{"gene":"POLR3G","run_date":"2026-04-28T19:45:44","timeline":{"discoveries":[{"year":1995,"finding":"The C31 subunit (yeast ortholog of POLR3G) is part of a three-subunit complex (C31, C34, C82) specific to RNA polymerase III. Partial deletion of the acidic C-terminus of C31 impairs transcription initiation of tRNA genes in the presence of general initiation factors (TFIIIB), but not non-specific transcription, termination, or recycling. Overexpression of the largest Pol III subunit (C160) suppresses the C31 truncation phenotype, indicating a functional interaction between C31 and C160.","method":"In vitro transcription assays, conditional yeast mutant (C-terminal deletion), suppressor overexpression, genetic epistasis","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1-2 — in vitro transcription assay + mutagenesis + genetic suppressor analysis in a single study with clear mechanistic readout","pmids":["7835345"],"is_preprint":false},{"year":2013,"finding":"POLR3G and the closely related POLR3GL subunit arose from a DNA-based gene duplication in a vertebrate ancestor. Both POLR3G-containing and POLR3GL-containing Pol III complexes occupy the same target genes genome-wide (shown by ChIP-seq), in constant proportions within a cell line, indicating equivalent target gene specificity. However, POLR3G-containing Pol III is relatively more abundant in dividing cells, and the POLR3G promoter—but not the POLR3GL promoter—binds the transcription factor MYC, establishing differential transcriptional regulation of the two subunit-encoding genes rather than neo-functionalization of the subunit products themselves.","method":"ChIP-seq, promoter analysis, transcription factor binding (MYC ChIP), quantitative comparison of Pol III isoform abundance in dividing vs. non-dividing cells","journal":"Genome research","confidence":"High","confidence_rationale":"Tier 2 — genome-wide ChIP-seq with multiple orthogonal approaches (ChIP, promoter analysis, quantitative isoform measurements); moderate-to-strong evidence from a single rigorous study","pmids":["24107381"],"is_preprint":false},{"year":2011,"finding":"POLR3G is required for maintenance of pluripotency in human embryonic stem cells (hESCs). Knockdown of POLR3G via inducible shRNA causes loss of pluripotency and promotes differentiation into all three germ layers without affecting apoptosis. Overexpression of POLR3G confers resistance to differentiation. POLR3G expression is regulated downstream of OCT4 and NANOG, and is modulated by the ERK1/2 signaling pathway.","method":"Inducible shRNA knockdown, overexpression, differentiation assays, pharmacological inhibition of ERK1/2, analysis of pluripotency markers","journal":"Stem cells (Dayton, Ohio)","confidence":"High","confidence_rationale":"Tier 2 — clean loss-of-function with defined cellular phenotype, gain-of-function, and pathway placement (OCT4/NANOG upstream, ERK1/2 regulation), multiple orthogonal readouts","pmids":["21898682"],"is_preprint":false},{"year":2020,"finding":"POLR3G and POLR3GL subunits can functionally compensate for each other in vivo: exogenous POLR3GL rescues the differentiation defect of POLR3G-knockout ESCs. Both Pol III isoforms bind the same target genes and exhibit equivalent functions in vitro and in vivo. POLR3G knockout mice die at a very early embryonic stage, while POLR3GL knockout mice survive embryogenesis but die ~3 weeks postnatally with growth defects and potential cerebellar neuronal defects, reflecting differential expression levels across developmental stages.","method":"Conditional and constitutive knockout mice, ESC differentiation rescue assays, in vitro transcription, ChIP","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1-2 — multiple genetic KO models combined with rescue experiments and in vitro assays; strong evidence from a single rigorous study","pmids":["32576691"],"is_preprint":false},{"year":2017,"finding":"POLR3G regulates a specific subset of the hPSC transcriptome including protein-coding genes, lincRNAs, microRNAs, and small nucleolar RNAs, and affects RNA splicing. Its primary function is in maintenance rather than repression of transcription. Among direct POLR3G targets, POLG (mitochondrial DNA polymerase gamma) is potentially important for sustaining stem cell status. The majority of POLR3G-regulated transcripts have pluripotency factor binding at their promoters.","method":"Deep-sequencing of polyA+ and smallRNA transcriptomes in POLR3G knockdown hPSCs, promoter analysis","journal":"Stem cell reports","confidence":"Medium","confidence_rationale":"Tier 2 — transcriptome-wide sequencing upon KD with defined cellular context, but mechanistic pathway placement is partially inferential","pmids":["28494942"],"is_preprint":false},{"year":2019,"finding":"In Xenopus (vertebrate model), a specific isoform of Polr3g (short form) is expressed early in the myogenic lineage. Forcing Polr3g expression during skeletal muscle differentiation causes a partial reversal of myogenic differentiation. Polr3g and the alternate isoform Polr3gL show distinct activity on tRNA isoacceptor synthesis as demonstrated by a custom tRNA microarray. Pol III-dependent transcripts, including tRNAs, are dramatically downregulated during skeletal muscle differentiation.","method":"Custom tRNA microarray, forced expression assay (overexpression in differentiating myoblasts), expression profiling during embryonic development","journal":"Developmental biology","confidence":"Medium","confidence_rationale":"Tier 2-3 — functional overexpression phenotype plus tRNA microarray in a vertebrate model; ortholog context consistent with mammalian POLR3G","pmids":["31173763"],"is_preprint":false},{"year":2022,"finding":"POLR3G knockout in the MDA-MB231 triple-negative breast cancer (TNBC) cell line dramatically reduces anchorage-independent growth and invasive capabilities in vitro, and impairs tumor growth and metastasis formation in orthotopic xenografts in vivo. POLR3G KO induces expression of the pioneer transcription factor FOXA1 and androgen receptor without altering EMT marker gene expression or proliferation.","method":"CRISPR KO, anchorage-independent growth assay, invasion assays, orthotopic xenograft, gene expression analysis","journal":"Cancers","confidence":"Medium","confidence_rationale":"Tier 2 — clean KO with defined in vitro and in vivo phenotypes plus gene expression mechanistic follow-up; single lab","pmids":["36497214"],"is_preprint":false},{"year":2023,"finding":"POLR3G promotes epithelial-mesenchymal transition (EMT) and invasion in bladder cancer cells via activation of the PI3K/AKT signaling pathway. Inhibition of PI3K/AKT with LY294002 reduces enhanced migration/invasion caused by POLR3G overexpression, and pathway activation with 740Y-P restores migration/invasion suppressed by POLR3G knockdown.","method":"shRNA knockdown, overexpression, in vitro migration/invasion assays, in vivo xenograft, pharmacological pathway inhibition/activation (LY294002, 740Y-P), Western blotting","journal":"FASEB journal","confidence":"Medium","confidence_rationale":"Tier 2-3 — pharmacological rescue/activation experiments place POLR3G upstream of PI3K/AKT; single lab","pmids":["37933949"],"is_preprint":false},{"year":2023,"finding":"POLR3G expression in cancer is controlled by a combinatorial regulatory platform: a gene-internal super-enhancer bound by multiple transcription factors (including MYC, ZNF131, ZNF207) drives expression, while gene-internal DNA methylation, retinoic acid-induced differentiation, and MXD4 negatively regulate it. Histone demethylase KDM5B is identified as a likely influencer of POLR3G gene activity. ZNF131 and ZNF207 functionally enhance POLR3G expression when overexpressed.","method":"Genomic survey of mRNA and chromatin signatures, functional TF overexpression assays, promoter/enhancer analysis, chromatin architecture analysis","journal":"Cancers","confidence":"Medium","confidence_rationale":"Tier 2-3 — functional TF overexpression experiments combined with large-scale genomic analysis; single lab, moderate mechanistic follow-up","pmids":["37894362"],"is_preprint":false},{"year":2023,"finding":"MicroRNA miR-26a-5p directly regulates POLR3G expression in lung cancer cells. Overexpression of miR-26a-5p reduces colony formation, sphere formation, and sensitizes lung cancer cells to paclitaxel, consistent with POLR3G being a functional target of miR-26a-5p in controlling cancer stemness.","method":"miRNA overexpression, luciferase reporter or direct target validation, colony/sphere formation assays, drug co-treatment","journal":"Non-coding RNA research","confidence":"Low","confidence_rationale":"Tier 3 — single lab, functional assay with limited mechanistic depth on the miR-POLR3G interaction itself","pmids":["36949748"],"is_preprint":false},{"year":2024,"finding":"POLR3G knockdown in bladder cancer cell lines inhibits proliferation, migration, and invasion. RNA sequencing of POLR3G knockdown cells reveals enrichment of differentially expressed genes in the Wnt and MAPK signaling pathways. Western blotting confirms downregulation of Wnt pathway proteins (Wnt5a/b, DVL2, LRP-6, phosphorylated LRP-6) upon POLR3G knockdown.","method":"shRNA knockdown, RNA sequencing, functional proliferation/migration/invasion assays, Western blotting, in vivo BBN-induced bladder cancer rat model","journal":"European journal of medical research","confidence":"Low","confidence_rationale":"Tier 3 — single lab, knockdown plus RNA-seq with Western blot confirmation; pathway placement partially inferential","pmids":["39039528"],"is_preprint":false}],"current_model":"POLR3G (RPC7α/C31) is a subunit unique to one of two forms of RNA polymerase III; it is part of a conserved C31-C34-C82 subcomplex that contacts TFIIIB to promote transcription initiation at tRNA and other Pol III target genes, is preferentially expressed in pluripotent/embryonic stem cells and proliferating/cancer cells under MYC-driven transcriptional control, is required for maintenance of pluripotent stem cell identity and early embryonic development, and its overexpression in cancers promotes invasion and metastasis through PI3K/AKT and Wnt/MAPK signaling pathways."},"narrative":{"teleology":[{"year":1995,"claim":"Establishing that the yeast C31 subunit (POLR3G ortholog) resides in a Pol III–specific subcomplex and is selectively required for transcription initiation rather than elongation, termination, or recycling answered how Pol III achieves promoter-dependent start-site selection.","evidence":"In vitro transcription assays with C-terminal truncation mutants and C160 suppressor overexpression in yeast","pmids":["7835345"],"confidence":"High","gaps":["Physical contacts between C31 and TFIIIB were inferred but not mapped at residue level","Whether the mammalian ortholog retains the same initiation-specific role was not tested"]},{"year":2011,"claim":"Demonstrating that POLR3G is required for pluripotency maintenance in human ESCs and is regulated by OCT4/NANOG/ERK1/2 connected a Pol III subunit to stem cell identity, revealing a cell-type-specific role beyond housekeeping transcription.","evidence":"Inducible shRNA knockdown and overexpression in hESCs with pluripotency marker and differentiation assays plus ERK1/2 pharmacological inhibition","pmids":["21898682"],"confidence":"High","gaps":["Whether pluripotency loss is caused by reduced tRNA synthesis or altered transcription of specific Pol III targets was unresolved","Downstream effectors mediating the differentiation phenotype were not identified"]},{"year":2013,"claim":"Genome-wide ChIP-seq showing POLR3G- and POLR3GL-containing Pol III occupy identical targets, combined with MYC-selective binding to the POLR3G promoter, established that paralog-specific biology arises from differential transcriptional regulation rather than functional divergence of the subunit proteins.","evidence":"ChIP-seq for both Pol III isoforms, MYC ChIP at POLR3G and POLR3GL promoters, quantitative isoform abundance in dividing vs. non-dividing cells","pmids":["24107381"],"confidence":"High","gaps":["Whether subtle target-gene-level quantitative differences exist between isoforms under specific conditions was not excluded","Post-transcriptional regulation of the two subunits was not examined"]},{"year":2017,"claim":"Transcriptomic profiling upon POLR3G knockdown in hPSCs revealed that POLR3G influences expression of protein-coding genes, lincRNAs, and snoRNAs beyond canonical Pol III targets, broadening its functional scope in stem cells.","evidence":"Deep sequencing of polyA+ and small RNA transcriptomes in POLR3G-depleted hPSCs","pmids":["28494942"],"confidence":"Medium","gaps":["Direct versus indirect transcriptional effects were not distinguished","Whether effects on splicing and non-coding RNA are mediated through altered tRNA pools or independent mechanisms is unknown"]},{"year":2019,"claim":"Studies in Xenopus showed Polr3g and Polr3gL have distinct effects on tRNA isoacceptor pools and that forced Polr3g expression partially reverses myogenic differentiation, extending the differentiation-linked role of POLR3G to a non-mammalian vertebrate.","evidence":"Custom tRNA microarray and overexpression in differentiating Xenopus myoblasts","pmids":["31173763"],"confidence":"Medium","gaps":["Mechanism by which altered tRNA isoacceptor ratios influence differentiation state was not established","Relevance of Xenopus findings to mammalian skeletal muscle differentiation not confirmed"]},{"year":2020,"claim":"Genetic knockout in mice proved that POLR3G is essential for early embryogenesis while POLR3GL supports later development, and rescue experiments in ESCs demonstrated functional interchangeability of the two subunits when co-expressed, resolving the question of intrinsic versus expression-dependent specificity.","evidence":"Constitutive and conditional KO mice, ESC differentiation rescue with exogenous POLR3GL, in vitro transcription and ChIP","pmids":["32576691"],"confidence":"High","gaps":["Why POLR3GL cannot compensate in early embryos despite biochemical equivalence (expression timing or level) was not fully dissected","Tissue-specific consequences of POLR3G loss in adult organs remain unexplored"]},{"year":2022,"claim":"CRISPR knockout of POLR3G in triple-negative breast cancer cells demonstrated a direct requirement for anchorage-independent growth, invasion, and in vivo metastasis, linking the Pol III subunit to cancer aggressiveness independently of proliferation rate changes.","evidence":"CRISPR KO in MDA-MB231, invasion assays, orthotopic xenograft model, gene expression profiling","pmids":["36497214"],"confidence":"Medium","gaps":["How POLR3G KO induces FOXA1 and androgen receptor expression mechanistically is unknown","Whether the metastasis phenotype depends on altered Pol III transcriptional output or a non-canonical function was not determined"]},{"year":2023,"claim":"Pharmacological rescue experiments placed POLR3G upstream of PI3K/AKT signaling in bladder cancer EMT, providing the first pathway-level mechanism for its pro-invasive effects, while parallel work mapped a gene-internal super-enhancer integrating MYC, ZNF131, ZNF207, and DNA methylation to control POLR3G expression in cancer.","evidence":"shRNA/overexpression with LY294002 and 740Y-P pathway modulation in bladder cancer cells; functional TF overexpression plus chromatin architecture analysis","pmids":["37933949","37894362"],"confidence":"Medium","gaps":["Intermediate steps between Pol III transcriptional output and PI3K/AKT activation remain unidentified","Whether the super-enhancer architecture is broadly conserved across cancer types is untested","KDM5B involvement is correlative, not functionally validated"]},{"year":2024,"claim":"RNA-seq upon POLR3G knockdown in bladder cancer implicated Wnt and MAPK pathway components as downstream effectors, expanding the signaling network influenced by POLR3G beyond PI3K/AKT.","evidence":"shRNA knockdown, RNA-seq, Western blot validation of Wnt pathway proteins in bladder cancer cells and BBN-induced rat model","pmids":["39039528"],"confidence":"Low","gaps":["Pathway placement is based on knockdown transcriptomics without epistasis or rescue experiments","Direct versus indirect regulation of Wnt/MAPK components is unresolved","Single-lab study without independent replication"]},{"year":null,"claim":"The mechanism by which altered Pol III transcriptional output (e.g., tRNA pools, other small non-coding RNAs) is transduced into activation of PI3K/AKT, Wnt/MAPK, or other oncogenic signaling pathways remains the central unresolved question.","evidence":"","pmids":[],"confidence":"Low","gaps":["No direct biochemical link between Pol III transcript changes and signaling pathway activation has been identified","Whether POLR3G has non-canonical, Pol III-independent functions is unexplored","Structural basis of the POLR3G–C34–C82 subcomplex interaction with TFIIIB in mammals lacks atomic-resolution data"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[0,2,4]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,1,2]}],"pathway":[{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[0,1,4]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[2,3,5]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[7]}],"complexes":["RNA polymerase III","C31-C34-C82 subcomplex"],"partners":["POLR3GL","MYC","ZNF131","ZNF207"],"other_free_text":[]},"mechanistic_narrative":"POLR3G encodes RPC7α, a subunit of one of two alternative forms of RNA polymerase III that functions within a conserved C31–C34–C82 subcomplex to promote transcription initiation at tRNA and other Pol III target genes [PMID:7835345]. POLR3G and its paralog POLR3GL arose by vertebrate gene duplication and assemble into Pol III complexes that occupy identical genomic targets, but POLR3G is preferentially expressed in pluripotent stem cells and proliferating/cancer cells owing to MYC-driven transcriptional activation of the POLR3G promoter and regulation downstream of OCT4/NANOG [PMID:24107381, PMID:21898682, PMID:37894362]. POLR3G is required for maintenance of pluripotency in human embryonic stem cells—its depletion triggers differentiation into all three germ layers—and POLR3G-knockout mice die at early embryonic stages, whereas the two isoform-containing Pol III complexes are functionally interchangeable when both subunits are co-expressed [PMID:21898682, PMID:32576691]. In cancer, POLR3G promotes invasion and metastasis in breast and bladder carcinoma models through engagement of PI3K/AKT and Wnt/MAPK signaling pathways [PMID:36497214, PMID:37933949, PMID:39039528]."},"prefetch_data":{"uniprot":{"accession":"O15318","full_name":"DNA-directed RNA polymerase III subunit RPC7","aliases":["DNA-directed RNA polymerase III subunit G","RNA polymerase III 32 kDa apha subunit","RPC32-alpha","RNA polymerase III 32 kDa subunit","RPC32"],"length_aa":223,"mass_kda":25.9,"function":"DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates (PubMed:20413673, PubMed:33558764, PubMed:34675218, PubMed:35637192). Specific peripheric component of RNA polymerase III (Pol III) which synthesizes small non-coding RNAs including 5S rRNA, snRNAs, tRNAs and miRNAs from at least 500 distinct genomic loci (PubMed:20154270, PubMed:20413673, PubMed:35637192). Acts as a long tether that bridges POLR3C/RPC3-POLR3F/RPC6-POLR3G/RPC7 heterotrimer and the mobile stalk of Pol III, coordinating the dynamics of Pol III stalk and clamp modules during the transition from apo to elongation state. Pol III exists as two alternative complexes defined by the mutually exclusive incorporation of subunit POLR3G/RPC7alpha or POLR3GL/RPC7beta. POLR3G/RPC7alpha modulates Pol III transcriptome by specifically enhancing the transcription of snaR-A non-coding RNAs. At resting state, occupies the active site of apo Pol III and keeps Pol III in an autoinhibitory mode, preventing non-specific transcription (PubMed:33558764, PubMed:33558766, PubMed:35637192). Pol III plays a key role in sensing and limiting infection by intracellular bacteria and DNA viruses. Acts as a nuclear and cytosolic DNA sensor involved in innate immune response. Can sense non-self dsDNA that serves as template for transcription into dsRNA. The non-self RNA polymerase III transcripts, such as Epstein-Barr virus-encoded RNAs (EBERs), induce type I interferon and NF-kappa-B through the RIG-I pathway (PubMed:19609254, PubMed:19631370)","subcellular_location":"Nucleus; Cytoplasm","url":"https://www.uniprot.org/uniprotkb/O15318/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/POLR3G","classification":"Not Classified","n_dependent_lines":15,"n_total_lines":1208,"dependency_fraction":0.012417218543046357},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000113356","cell_line_id":"CID000715","localizations":[{"compartment":"nuclear_punctae","grade":3},{"compartment":"nucleoplasm","grade":2}],"interactors":[{"gene":"POLR1C","stoichiometry":10.0},{"gene":"POLR1D","stoichiometry":10.0},{"gene":"POLR2E","stoichiometry":10.0},{"gene":"POLR2F","stoichiometry":10.0},{"gene":"POLR2H","stoichiometry":10.0},{"gene":"POLR2K","stoichiometry":10.0},{"gene":"POLR3A","stoichiometry":10.0},{"gene":"POLR3B","stoichiometry":10.0},{"gene":"POLR3C","stoichiometry":10.0},{"gene":"POLR3D","stoichiometry":10.0}],"url":"https://opencell.sf.czbiohub.org/target/CID000715","total_profiled":1310},"omim":[{"mim_id":"617457","title":"POLYMERASE III, RNA, SUBUNIT G-LIKE; POLR3GL","url":"https://www.omim.org/entry/617457"},{"mim_id":"617456","title":"POLYMERASE III, RNA, SUBUNIT G; POLR3G","url":"https://www.omim.org/entry/617456"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Nuclear bodies","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/POLR3G"},"hgnc":{"alias_symbol":["RPC32","RPC7","C31"],"prev_symbol":[]},"alphafold":{"accession":"O15318","domains":[{"cath_id":"-","chopping":"48-76_90-101","consensus_level":"medium","plddt":87.8922,"start":48,"end":101}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O15318","model_url":"https://alphafold.ebi.ac.uk/files/AF-O15318-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O15318-F1-predicted_aligned_error_v6.png","plddt_mean":67.31},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=POLR3G","jax_strain_url":"https://www.jax.org/strain/search?query=POLR3G"},"sequence":{"accession":"O15318","fasta_url":"https://rest.uniprot.org/uniprotkb/O15318.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O15318/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O15318"}},"corpus_meta":[{"pmid":"7972041","id":"PMC_7972041","title":"Prostate and mammary adenocarcinoma in transgenic mice carrying a rat C3(1) simian virus 40 large tumor antigen fusion gene.","date":"1994","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/7972041","citation_count":306,"is_preprint":false},{"pmid":"1762157","id":"PMC_1762157","title":"Analysis of the integration function of the streptomycete bacteriophage phi C31.","date":"1991","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/1762157","citation_count":168,"is_preprint":false},{"pmid":"5014931","id":"PMC_5014931","title":"Characterization of temperate actinophage phi C31 isolated from Streptomyces coelicolor A3(2).","date":"1972","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/5014931","citation_count":161,"is_preprint":false},{"pmid":"2818590","id":"PMC_2818590","title":"Functional characterization of an androgen response element in the first intron of the C3(1) gene of prostatic binding protein.","date":"1989","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/2818590","citation_count":123,"is_preprint":false},{"pmid":"6292047","id":"PMC_6292047","title":"The expression of Streptomyces and Escherichia coli drug-resistance determinants cloned into the Streptomyces phage phi C31.","date":"1982","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/6292047","citation_count":112,"is_preprint":false},{"pmid":"12270833","id":"PMC_12270833","title":"The streptomyces genome contains multiple pseudo-attB sites for the (phi)C31-encoded site-specific recombination system.","date":"2002","source":"Journal of bacteriology","url":"https://pubmed.ncbi.nlm.nih.gov/12270833","citation_count":109,"is_preprint":false},{"pmid":"16550351","id":"PMC_16550351","title":"Maternal transfer of complement components C3-1, C3-3, C3-4, C4, C5, C7, Bf, and Df to offspring in rainbow trout (Oncorhynchus mykiss).","date":"2006","source":"Immunogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/16550351","citation_count":93,"is_preprint":false},{"pmid":"7835345","id":"PMC_7835345","title":"A mutation in the C31 subunit of Saccharomyces cerevisiae RNA polymerase III affects transcription initiation.","date":"1995","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/7835345","citation_count":90,"is_preprint":false},{"pmid":"2989111","id":"PMC_2989111","title":"New derivatives of the Streptomyces temperate phage phi C31 useful for the cloning and functional analysis of Streptomyces DNA.","date":"1985","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/2989111","citation_count":63,"is_preprint":false},{"pmid":"16672982","id":"PMC_16672982","title":"Phi c31 integrase induces chromosomal aberrations in primary human fibroblasts.","date":"2006","source":"Gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/16672982","citation_count":62,"is_preprint":false},{"pmid":"22045025","id":"PMC_22045025","title":"Consumption of high ω-3 fatty acid diet suppressed prostate tumorigenesis in C3(1) Tag mice.","date":"2011","source":"Carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/22045025","citation_count":59,"is_preprint":false},{"pmid":"1656389","id":"PMC_1656389","title":"Structural analysis of the actinophage phi C31 attachment site.","date":"1991","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/1656389","citation_count":58,"is_preprint":false},{"pmid":"17407311","id":"PMC_17407311","title":"Breast cancer prevention by green tea catechins and black tea theaflavins in the C3(1) SV40 T,t antigen transgenic mouse model is accompanied by increased apoptosis and a decrease in oxidative DNA adducts.","date":"2007","source":"Journal of agricultural and food chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17407311","citation_count":50,"is_preprint":false},{"pmid":"8224520","id":"PMC_8224520","title":"Proteins interacting with an androgen-responsive unit in the C3(1) gene intron.","date":"1993","source":"Molecular and cellular endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/8224520","citation_count":47,"is_preprint":false},{"pmid":"24107381","id":"PMC_24107381","title":"Gene duplication and neofunctionalization: POLR3G and POLR3GL.","date":"2013","source":"Genome research","url":"https://pubmed.ncbi.nlm.nih.gov/24107381","citation_count":46,"is_preprint":false},{"pmid":"19679105","id":"PMC_19679105","title":"Mechanism of cytotoxicity mediated by the C31 fragment of the amyloid precursor protein.","date":"2009","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/19679105","citation_count":46,"is_preprint":false},{"pmid":"7600815","id":"PMC_7600815","title":"Morphology, histology, and ultrastructure of human C31 organ-cultured corneas.","date":"1995","source":"Cornea","url":"https://pubmed.ncbi.nlm.nih.gov/7600815","citation_count":42,"is_preprint":false},{"pmid":"32083668","id":"PMC_32083668","title":"A memory switch for plant synthetic biology based on the phage ϕC31 integration system.","date":"2020","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/32083668","citation_count":41,"is_preprint":false},{"pmid":"12431051","id":"PMC_12431051","title":"Synthesis and evaluation of indenopyrazoles as cyclin-dependent kinase inhibitors. 3. Structure activity relationships at C3(1,2).","date":"2002","source":"Journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12431051","citation_count":40,"is_preprint":false},{"pmid":"21898682","id":"PMC_21898682","title":"A novel role for an RNA polymerase III subunit POLR3G in regulating pluripotency in human embryonic stem cells.","date":"2011","source":"Stem cells (Dayton, Ohio)","url":"https://pubmed.ncbi.nlm.nih.gov/21898682","citation_count":40,"is_preprint":false},{"pmid":"23150546","id":"PMC_23150546","title":"Gated rotation mechanism of site-specific recombination by ϕC31 integrase.","date":"2012","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/23150546","citation_count":39,"is_preprint":false},{"pmid":"21774594","id":"PMC_21774594","title":"Dietary walnut suppressed mammary gland tumorigenesis in the C(3)1 TAg mouse.","date":"2011","source":"Nutrition and cancer","url":"https://pubmed.ncbi.nlm.nih.gov/21774594","citation_count":39,"is_preprint":false},{"pmid":"3185504","id":"PMC_3185504","title":"The repressor gene (c) of the Streptomyces temperate phage phi c31: nucleotide sequence, analysis and functional cloning.","date":"1988","source":"Molecular & general genetics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/3185504","citation_count":34,"is_preprint":false},{"pmid":"6307817","id":"PMC_6307817","title":"The restriction mapping of c gene deletions in Streptomyces bacteriophage phi C31 and their use in cloning vector development.","date":"1983","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/6307817","citation_count":32,"is_preprint":false},{"pmid":"7642495","id":"PMC_7642495","title":"Two genes involved in the phase-variable phi C31 resistance mechanism of Streptomyces coelicolor A3(2).","date":"1995","source":"Journal of bacteriology","url":"https://pubmed.ncbi.nlm.nih.gov/7642495","citation_count":32,"is_preprint":false},{"pmid":"6269960","id":"PMC_6269960","title":"Restriction mapping of the DNA of the Streptomyces temperate phage phi C31 and its derivatives.","date":"1981","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/6269960","citation_count":27,"is_preprint":false},{"pmid":"25101216","id":"PMC_25101216","title":"Inactivation of Streptomyces phage ɸC31 by 405 nm light: Requirement for exogenous photosensitizers?","date":"2014","source":"Bacteriophage","url":"https://pubmed.ncbi.nlm.nih.gov/25101216","citation_count":27,"is_preprint":false},{"pmid":"2040440","id":"PMC_2040440","title":"Construction and transduction of a shuttle vector bearing the cos site of Streptomyces phage phi C31 and determination of its cohesive ends.","date":"1991","source":"FEMS microbiology letters","url":"https://pubmed.ncbi.nlm.nih.gov/2040440","citation_count":27,"is_preprint":false},{"pmid":"32576691","id":"PMC_32576691","title":"Functions of paralogous RNA polymerase III subunits POLR3G and POLR3GL in mouse development.","date":"2020","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/32576691","citation_count":26,"is_preprint":false},{"pmid":"11532128","id":"PMC_11532128","title":"Glycosylation of a Streptomyces coelicolor A3(2) cell envelope protein is required for infection by bacteriophage phi C31.","date":"2001","source":"Molecular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/11532128","citation_count":24,"is_preprint":false},{"pmid":"6271645","id":"PMC_6271645","title":"Dispensable sequences and packaging constraints of DNA from the Streptomyces temperate phage phi C31.","date":"1981","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/6271645","citation_count":23,"is_preprint":false},{"pmid":"37933949","id":"PMC_37933949","title":"POLR3G promotes EMT via PI3K/AKT signaling pathway in bladder cancer.","date":"2023","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/37933949","citation_count":21,"is_preprint":false},{"pmid":"8446035","id":"PMC_8446035","title":"Genetic analysis of the phi C31-specific phage growth limitation (Pgl) system of Streptomyces coelicolor A3(2).","date":"1993","source":"Molecular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/8446035","citation_count":19,"is_preprint":false},{"pmid":"28494942","id":"PMC_28494942","title":"RNA Polymerase III Subunit POLR3G Regulates Specific Subsets of PolyA+ and SmallRNA Transcriptomes and Splicing in Human Pluripotent Stem Cells.","date":"2017","source":"Stem cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/28494942","citation_count":19,"is_preprint":false},{"pmid":"1779769","id":"PMC_1779769","title":"Three in-frame N-terminally different proteins are produced from the repressor locus of the Streptomyces bacteriophage phi C31.","date":"1991","source":"Molecular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/1779769","citation_count":19,"is_preprint":false},{"pmid":"27387286","id":"PMC_27387286","title":"The mechanism of ϕC31 integrase directionality: experimental analysis and computational modelling.","date":"2016","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/27387286","citation_count":18,"is_preprint":false},{"pmid":"31469960","id":"PMC_31469960","title":"C30 and C31 Triterpenoids and Triterpene Sugar Esters with Cytotoxic Activities from Edible Mushroom Fomitopsis pinicola (Sw. Ex Fr.) Krast.","date":"2019","source":"Journal of agricultural and food chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/31469960","citation_count":18,"is_preprint":false},{"pmid":"23435975","id":"PMC_23435975","title":"FK506 maturation involves a cytochrome p450 protein-catalyzed four-electron C-9 oxidation in parallel with a C-31 O-methylation.","date":"2013","source":"Journal of bacteriology","url":"https://pubmed.ncbi.nlm.nih.gov/23435975","citation_count":18,"is_preprint":false},{"pmid":"29228292","id":"PMC_29228292","title":"Recombination directionality factor gp3 binds ϕC31 integrase via the zinc domain, potentially affecting the trajectory of the coiled-coil motif.","date":"2018","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/29228292","citation_count":16,"is_preprint":false},{"pmid":"7651131","id":"PMC_7651131","title":"Control of lytic development in the Streptomyces temperate phage phi C31.","date":"1995","source":"Molecular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/7651131","citation_count":16,"is_preprint":false},{"pmid":"29715599","id":"PMC_29715599","title":"Lanostane-type C31 triterpenoid derivatives from the fruiting bodies of cultivated Fomitopsis palustris.","date":"2018","source":"Phytochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29715599","citation_count":16,"is_preprint":false},{"pmid":"12374845","id":"PMC_12374845","title":"A gene encoding a homologue of dolichol phosphate-beta-D-mannose synthase is required for infection of Streptomyces coelicolor A3(2) by phage (phi)C31.","date":"2002","source":"Journal of bacteriology","url":"https://pubmed.ncbi.nlm.nih.gov/12374845","citation_count":16,"is_preprint":false},{"pmid":"36497214","id":"PMC_36497214","title":"The POLR3G Subunit of Human RNA Polymerase III Regulates Tumorigenesis and Metastasis in Triple-Negative Breast Cancer.","date":"2022","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/36497214","citation_count":15,"is_preprint":false},{"pmid":"30169869","id":"PMC_30169869","title":"Protein Phosphatase 1-Targeting Small-Molecule C31 Inhibits Ebola Virus Replication.","date":"2018","source":"The Journal of infectious diseases","url":"https://pubmed.ncbi.nlm.nih.gov/30169869","citation_count":15,"is_preprint":false},{"pmid":"23574114","id":"PMC_23574114","title":"Site-specific T-DNA integration in Arabidopsis thaliana mediated by the combined action of CRE recombinase and ϕC31 integrase.","date":"2013","source":"The Plant journal : for cell and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/23574114","citation_count":15,"is_preprint":false},{"pmid":"27986956","id":"PMC_27986956","title":"Single-molecule analysis of ϕC31 integrase-mediated site-specific recombination by tethered particle motion.","date":"2016","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/27986956","citation_count":15,"is_preprint":false},{"pmid":"3806054","id":"PMC_3806054","title":"Characteristics of the developmental cycle of actinophage phi C31.","date":"1986","source":"Journal of general microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/3806054","citation_count":15,"is_preprint":false},{"pmid":"17249800","id":"PMC_17249800","title":"Development of an access route to the c31-c52 central core of amphidinol 3.","date":"2007","source":"Organic letters","url":"https://pubmed.ncbi.nlm.nih.gov/17249800","citation_count":15,"is_preprint":false},{"pmid":"31667500","id":"PMC_31667500","title":"Control of ϕC31 integrase-mediated site-specific recombination by protein trans-splicing.","date":"2019","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/31667500","citation_count":14,"is_preprint":false},{"pmid":"7106562","id":"PMC_7106562","title":"Sequence organization of a viral DNA insertion present in the adenovirus-type-5-transformed hamster line BHK268-C31.","date":"1982","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/7106562","citation_count":14,"is_preprint":false},{"pmid":"1479345","id":"PMC_1479345","title":"Global transcription pattern of phi C31 after induction of a Streptomyces coelicolor lysogen at different growth stages.","date":"1992","source":"Journal of general microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/1479345","citation_count":13,"is_preprint":false},{"pmid":"9472712","id":"PMC_9472712","title":"Altered expression of transforming growth factor betas during urethral and bulbourethral gland tumor progression in transgenic mice carrying the androgen-responsive C3(1) 5' flanking region fused to SV40 large T antigen.","date":"1998","source":"Carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/9472712","citation_count":13,"is_preprint":false},{"pmid":"2628169","id":"PMC_2628169","title":"Transcriptional analysis of the repressor gene of the temperate Streptomyces phage phi C31.","date":"1989","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/2628169","citation_count":13,"is_preprint":false},{"pmid":"36949748","id":"PMC_36949748","title":"Regulating POLR3G by MicroRNA-26a-5p as a promising therapeutic target of lung cancer stemness and chemosensitivity.","date":"2023","source":"Non-coding RNA research","url":"https://pubmed.ncbi.nlm.nih.gov/36949748","citation_count":12,"is_preprint":false},{"pmid":"7934940","id":"PMC_7934940","title":"Multiple novel promoters from the early region in the Streptomyces temperate phage phi C31 are activated during lytic development.","date":"1993","source":"Molecular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/7934940","citation_count":12,"is_preprint":false},{"pmid":"8088546","id":"PMC_8088546","title":"Sequence of the essential early region of phi C31, a temperate phage of Streptomyces spp. with unusual features in its lytic development.","date":"1994","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/8088546","citation_count":12,"is_preprint":false},{"pmid":"1708439","id":"PMC_1708439","title":"Induction of a phi C31 prophage inhibits rRNA transcription in Streptomyces coelicolor A3(2).","date":"1990","source":"Molecular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/1708439","citation_count":11,"is_preprint":false},{"pmid":"1452040","id":"PMC_1452040","title":"Transcription map of the early region of the Streptomyces bacteriophage phi C31.","date":"1992","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/1452040","citation_count":11,"is_preprint":false},{"pmid":"37949680","id":"PMC_37949680","title":"APP-C31: An Intracellular Promoter of Both Metal-Free and Metal-Bound Amyloid-β40 Aggregation and Toxicity in Alzheimer's Disease.","date":"2023","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/37949680","citation_count":10,"is_preprint":false},{"pmid":"31173763","id":"PMC_31173763","title":"Skeletal muscle differentiation drives a dramatic downregulation of RNA polymerase III activity and differential expression of Polr3g isoforms.","date":"2019","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/31173763","citation_count":10,"is_preprint":false},{"pmid":"20158522","id":"PMC_20158522","title":"Gene targeting for O-methyltransferase genes, mycE and mycF, on the chromosome of Micromonospora griseorubida producing mycinamicin with a disruption cassette containing the bacteriophage phi C31 attB attachment site.","date":"2010","source":"FEMS microbiology letters","url":"https://pubmed.ncbi.nlm.nih.gov/20158522","citation_count":10,"is_preprint":false},{"pmid":"19745601","id":"PMC_19745601","title":"Optimization of Streptomyces bacteriophage phi C31 integrase system to prevent post integrative gene silencing in pulmonary type II cells.","date":"2009","source":"Experimental & molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/19745601","citation_count":7,"is_preprint":false},{"pmid":"20578458","id":"PMC_20578458","title":"Intramolecular integration assay validates integrase phi C31 and R4 potential in a variety of insect cells.","date":"2009","source":"The Southeast Asian journal of tropical medicine and public health","url":"https://pubmed.ncbi.nlm.nih.gov/20578458","citation_count":7,"is_preprint":false},{"pmid":"36710885","id":"PMC_36710885","title":"RNA polymerase III transcription and cancer: A tale of two RPC7 subunits.","date":"2023","source":"Frontiers in molecular biosciences","url":"https://pubmed.ncbi.nlm.nih.gov/36710885","citation_count":6,"is_preprint":false},{"pmid":"39487291","id":"PMC_39487291","title":"Variable orthogonality of serine integrase interactions within the ϕC31 family.","date":"2024","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/39487291","citation_count":6,"is_preprint":false},{"pmid":"6947925","id":"PMC_6947925","title":"[Genetic mapping and characteristics of the actinophage phi C31 deletion mutants of Streptomyces coelicolor A3(2) incapable of lysogenization].","date":"1981","source":"Genetika","url":"https://pubmed.ncbi.nlm.nih.gov/6947925","citation_count":5,"is_preprint":false},{"pmid":"15911130","id":"PMC_15911130","title":"C31 enhances voltage-gated calcium channel currents in undifferentiated PC12 cells.","date":"2005","source":"Neuroscience letters","url":"https://pubmed.ncbi.nlm.nih.gov/15911130","citation_count":5,"is_preprint":false},{"pmid":"37894362","id":"PMC_37894362","title":"A Combinatorial Regulatory Platform Determines Expression of RNA Polymerase III Subunit RPC7α (POLR3G) in Cancer.","date":"2023","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/37894362","citation_count":4,"is_preprint":false},{"pmid":"36269749","id":"PMC_36269749","title":"Mammary Tumor Growth and Proliferation Are Dependent on Growth Hormone in Female SV40 C3(1) T-Antigen Mice.","date":"2022","source":"Endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/36269749","citation_count":4,"is_preprint":false},{"pmid":"33817748","id":"PMC_33817748","title":"Maternal fish oil consumption has a negative impact on mammary gland tumorigenesis in C3(1) Tag mice offspring.","date":"2021","source":"European journal of nutrition","url":"https://pubmed.ncbi.nlm.nih.gov/33817748","citation_count":4,"is_preprint":false},{"pmid":"38617232","id":"PMC_38617232","title":"Variable orthogonality of RDF - large serine integrase interactions within the ϕC31 family.","date":"2024","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/38617232","citation_count":2,"is_preprint":false},{"pmid":"39039528","id":"PMC_39039528","title":"The upregulation of POLR3G correlates with increased malignancy of bladder urothelium.","date":"2024","source":"European journal of medical research","url":"https://pubmed.ncbi.nlm.nih.gov/39039528","citation_count":1,"is_preprint":false},{"pmid":"7857157","id":"PMC_7857157","title":"[Actinomycete plasmid and integrative vectors based on DNA of the temperate Phi C31 actinophage, determining limitation of lytic development of phage Phi C31, not dependent on repressor].","date":"1994","source":"Antibiotiki i khimioterapiia = Antibiotics and chemoterapy [sic]","url":"https://pubmed.ncbi.nlm.nih.gov/7857157","citation_count":1,"is_preprint":false},{"pmid":"39559319","id":"PMC_39559319","title":"NMI, POLR3G and APIP are the key molecules connecting glaucoma with high intraocular pressure: a clue for early diagnostic biomarker candidates.","date":"2024","source":"International journal of ophthalmology","url":"https://pubmed.ncbi.nlm.nih.gov/39559319","citation_count":0,"is_preprint":false},{"pmid":"41639863","id":"PMC_41639863","title":"APP-C31 pathology as a target in neurodegenerative diseases.","date":"2026","source":"Journal of biomedical science","url":"https://pubmed.ncbi.nlm.nih.gov/41639863","citation_count":0,"is_preprint":false},{"pmid":"6289084","id":"PMC_6289084","title":"[Physical mapping of actinophage Streptomyces coelicolor A3(2). VI. The use of deletion mutants of actinophage phi C31 for construction of phage vectors].","date":"1982","source":"Molekuliarnaia biologiia","url":"https://pubmed.ncbi.nlm.nih.gov/6289084","citation_count":0,"is_preprint":false},{"pmid":"6291223","id":"PMC_6291223","title":"[Extraction and freeze-drying of herpes virus C3-1 isolated from turkeys].","date":"1982","source":"Veterinarno-meditsinski nauki","url":"https://pubmed.ncbi.nlm.nih.gov/6291223","citation_count":0,"is_preprint":false},{"pmid":"3270545","id":"PMC_3270545","title":"The effect of rifampicin on the development of the Streptomyces bacteriophage phi C31.","date":"1988","source":"Microbiologia (Madrid, Spain)","url":"https://pubmed.ncbi.nlm.nih.gov/3270545","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.07.08.602524","title":"Plasmid2MC: Efficient cell-free recombination of plasmids into high-purity minicircle DNA for use in genome editing applications","date":"2024-07-08","source":"bioRxiv","url":"https://doi.org/10.1101/2024.07.08.602524","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":33908,"output_tokens":3109,"usd":0.074179},"stage2":{"model":"claude-opus-4-6","input_tokens":6445,"output_tokens":2738,"usd":0.151012},"total_usd":0.225191,"stage1_batch_id":"msgbatch_01SUR8NDzVrNmyszWyJeCct7","stage2_batch_id":"msgbatch_01G7Yfutko3cDWzejrwH5AMx","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1995,\n      \"finding\": \"The C31 subunit (yeast ortholog of POLR3G) is part of a three-subunit complex (C31, C34, C82) specific to RNA polymerase III. Partial deletion of the acidic C-terminus of C31 impairs transcription initiation of tRNA genes in the presence of general initiation factors (TFIIIB), but not non-specific transcription, termination, or recycling. Overexpression of the largest Pol III subunit (C160) suppresses the C31 truncation phenotype, indicating a functional interaction between C31 and C160.\",\n      \"method\": \"In vitro transcription assays, conditional yeast mutant (C-terminal deletion), suppressor overexpression, genetic epistasis\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — in vitro transcription assay + mutagenesis + genetic suppressor analysis in a single study with clear mechanistic readout\",\n      \"pmids\": [\"7835345\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"POLR3G and the closely related POLR3GL subunit arose from a DNA-based gene duplication in a vertebrate ancestor. Both POLR3G-containing and POLR3GL-containing Pol III complexes occupy the same target genes genome-wide (shown by ChIP-seq), in constant proportions within a cell line, indicating equivalent target gene specificity. However, POLR3G-containing Pol III is relatively more abundant in dividing cells, and the POLR3G promoter—but not the POLR3GL promoter—binds the transcription factor MYC, establishing differential transcriptional regulation of the two subunit-encoding genes rather than neo-functionalization of the subunit products themselves.\",\n      \"method\": \"ChIP-seq, promoter analysis, transcription factor binding (MYC ChIP), quantitative comparison of Pol III isoform abundance in dividing vs. non-dividing cells\",\n      \"journal\": \"Genome research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genome-wide ChIP-seq with multiple orthogonal approaches (ChIP, promoter analysis, quantitative isoform measurements); moderate-to-strong evidence from a single rigorous study\",\n      \"pmids\": [\"24107381\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"POLR3G is required for maintenance of pluripotency in human embryonic stem cells (hESCs). Knockdown of POLR3G via inducible shRNA causes loss of pluripotency and promotes differentiation into all three germ layers without affecting apoptosis. Overexpression of POLR3G confers resistance to differentiation. POLR3G expression is regulated downstream of OCT4 and NANOG, and is modulated by the ERK1/2 signaling pathway.\",\n      \"method\": \"Inducible shRNA knockdown, overexpression, differentiation assays, pharmacological inhibition of ERK1/2, analysis of pluripotency markers\",\n      \"journal\": \"Stem cells (Dayton, Ohio)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean loss-of-function with defined cellular phenotype, gain-of-function, and pathway placement (OCT4/NANOG upstream, ERK1/2 regulation), multiple orthogonal readouts\",\n      \"pmids\": [\"21898682\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"POLR3G and POLR3GL subunits can functionally compensate for each other in vivo: exogenous POLR3GL rescues the differentiation defect of POLR3G-knockout ESCs. Both Pol III isoforms bind the same target genes and exhibit equivalent functions in vitro and in vivo. POLR3G knockout mice die at a very early embryonic stage, while POLR3GL knockout mice survive embryogenesis but die ~3 weeks postnatally with growth defects and potential cerebellar neuronal defects, reflecting differential expression levels across developmental stages.\",\n      \"method\": \"Conditional and constitutive knockout mice, ESC differentiation rescue assays, in vitro transcription, ChIP\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — multiple genetic KO models combined with rescue experiments and in vitro assays; strong evidence from a single rigorous study\",\n      \"pmids\": [\"32576691\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"POLR3G regulates a specific subset of the hPSC transcriptome including protein-coding genes, lincRNAs, microRNAs, and small nucleolar RNAs, and affects RNA splicing. Its primary function is in maintenance rather than repression of transcription. Among direct POLR3G targets, POLG (mitochondrial DNA polymerase gamma) is potentially important for sustaining stem cell status. The majority of POLR3G-regulated transcripts have pluripotency factor binding at their promoters.\",\n      \"method\": \"Deep-sequencing of polyA+ and smallRNA transcriptomes in POLR3G knockdown hPSCs, promoter analysis\",\n      \"journal\": \"Stem cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — transcriptome-wide sequencing upon KD with defined cellular context, but mechanistic pathway placement is partially inferential\",\n      \"pmids\": [\"28494942\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"In Xenopus (vertebrate model), a specific isoform of Polr3g (short form) is expressed early in the myogenic lineage. Forcing Polr3g expression during skeletal muscle differentiation causes a partial reversal of myogenic differentiation. Polr3g and the alternate isoform Polr3gL show distinct activity on tRNA isoacceptor synthesis as demonstrated by a custom tRNA microarray. Pol III-dependent transcripts, including tRNAs, are dramatically downregulated during skeletal muscle differentiation.\",\n      \"method\": \"Custom tRNA microarray, forced expression assay (overexpression in differentiating myoblasts), expression profiling during embryonic development\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — functional overexpression phenotype plus tRNA microarray in a vertebrate model; ortholog context consistent with mammalian POLR3G\",\n      \"pmids\": [\"31173763\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"POLR3G knockout in the MDA-MB231 triple-negative breast cancer (TNBC) cell line dramatically reduces anchorage-independent growth and invasive capabilities in vitro, and impairs tumor growth and metastasis formation in orthotopic xenografts in vivo. POLR3G KO induces expression of the pioneer transcription factor FOXA1 and androgen receptor without altering EMT marker gene expression or proliferation.\",\n      \"method\": \"CRISPR KO, anchorage-independent growth assay, invasion assays, orthotopic xenograft, gene expression analysis\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KO with defined in vitro and in vivo phenotypes plus gene expression mechanistic follow-up; single lab\",\n      \"pmids\": [\"36497214\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"POLR3G promotes epithelial-mesenchymal transition (EMT) and invasion in bladder cancer cells via activation of the PI3K/AKT signaling pathway. Inhibition of PI3K/AKT with LY294002 reduces enhanced migration/invasion caused by POLR3G overexpression, and pathway activation with 740Y-P restores migration/invasion suppressed by POLR3G knockdown.\",\n      \"method\": \"shRNA knockdown, overexpression, in vitro migration/invasion assays, in vivo xenograft, pharmacological pathway inhibition/activation (LY294002, 740Y-P), Western blotting\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — pharmacological rescue/activation experiments place POLR3G upstream of PI3K/AKT; single lab\",\n      \"pmids\": [\"37933949\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"POLR3G expression in cancer is controlled by a combinatorial regulatory platform: a gene-internal super-enhancer bound by multiple transcription factors (including MYC, ZNF131, ZNF207) drives expression, while gene-internal DNA methylation, retinoic acid-induced differentiation, and MXD4 negatively regulate it. Histone demethylase KDM5B is identified as a likely influencer of POLR3G gene activity. ZNF131 and ZNF207 functionally enhance POLR3G expression when overexpressed.\",\n      \"method\": \"Genomic survey of mRNA and chromatin signatures, functional TF overexpression assays, promoter/enhancer analysis, chromatin architecture analysis\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — functional TF overexpression experiments combined with large-scale genomic analysis; single lab, moderate mechanistic follow-up\",\n      \"pmids\": [\"37894362\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"MicroRNA miR-26a-5p directly regulates POLR3G expression in lung cancer cells. Overexpression of miR-26a-5p reduces colony formation, sphere formation, and sensitizes lung cancer cells to paclitaxel, consistent with POLR3G being a functional target of miR-26a-5p in controlling cancer stemness.\",\n      \"method\": \"miRNA overexpression, luciferase reporter or direct target validation, colony/sphere formation assays, drug co-treatment\",\n      \"journal\": \"Non-coding RNA research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single lab, functional assay with limited mechanistic depth on the miR-POLR3G interaction itself\",\n      \"pmids\": [\"36949748\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"POLR3G knockdown in bladder cancer cell lines inhibits proliferation, migration, and invasion. RNA sequencing of POLR3G knockdown cells reveals enrichment of differentially expressed genes in the Wnt and MAPK signaling pathways. Western blotting confirms downregulation of Wnt pathway proteins (Wnt5a/b, DVL2, LRP-6, phosphorylated LRP-6) upon POLR3G knockdown.\",\n      \"method\": \"shRNA knockdown, RNA sequencing, functional proliferation/migration/invasion assays, Western blotting, in vivo BBN-induced bladder cancer rat model\",\n      \"journal\": \"European journal of medical research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single lab, knockdown plus RNA-seq with Western blot confirmation; pathway placement partially inferential\",\n      \"pmids\": [\"39039528\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"POLR3G (RPC7α/C31) is a subunit unique to one of two forms of RNA polymerase III; it is part of a conserved C31-C34-C82 subcomplex that contacts TFIIIB to promote transcription initiation at tRNA and other Pol III target genes, is preferentially expressed in pluripotent/embryonic stem cells and proliferating/cancer cells under MYC-driven transcriptional control, is required for maintenance of pluripotent stem cell identity and early embryonic development, and its overexpression in cancers promotes invasion and metastasis through PI3K/AKT and Wnt/MAPK signaling pathways.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"POLR3G encodes RPC7α, a subunit of one of two alternative forms of RNA polymerase III that functions within a conserved C31–C34–C82 subcomplex to promote transcription initiation at tRNA and other Pol III target genes [PMID:7835345]. POLR3G and its paralog POLR3GL arose by vertebrate gene duplication and assemble into Pol III complexes that occupy identical genomic targets, but POLR3G is preferentially expressed in pluripotent stem cells and proliferating/cancer cells owing to MYC-driven transcriptional activation of the POLR3G promoter and regulation downstream of OCT4/NANOG [PMID:24107381, PMID:21898682, PMID:37894362]. POLR3G is required for maintenance of pluripotency in human embryonic stem cells—its depletion triggers differentiation into all three germ layers—and POLR3G-knockout mice die at early embryonic stages, whereas the two isoform-containing Pol III complexes are functionally interchangeable when both subunits are co-expressed [PMID:21898682, PMID:32576691]. In cancer, POLR3G promotes invasion and metastasis in breast and bladder carcinoma models through engagement of PI3K/AKT and Wnt/MAPK signaling pathways [PMID:36497214, PMID:37933949, PMID:39039528].\",\n  \"teleology\": [\n    {\n      \"year\": 1995,\n      \"claim\": \"Establishing that the yeast C31 subunit (POLR3G ortholog) resides in a Pol III–specific subcomplex and is selectively required for transcription initiation rather than elongation, termination, or recycling answered how Pol III achieves promoter-dependent start-site selection.\",\n      \"evidence\": \"In vitro transcription assays with C-terminal truncation mutants and C160 suppressor overexpression in yeast\",\n      \"pmids\": [\"7835345\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Physical contacts between C31 and TFIIIB were inferred but not mapped at residue level\",\n        \"Whether the mammalian ortholog retains the same initiation-specific role was not tested\"\n      ]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Demonstrating that POLR3G is required for pluripotency maintenance in human ESCs and is regulated by OCT4/NANOG/ERK1/2 connected a Pol III subunit to stem cell identity, revealing a cell-type-specific role beyond housekeeping transcription.\",\n      \"evidence\": \"Inducible shRNA knockdown and overexpression in hESCs with pluripotency marker and differentiation assays plus ERK1/2 pharmacological inhibition\",\n      \"pmids\": [\"21898682\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether pluripotency loss is caused by reduced tRNA synthesis or altered transcription of specific Pol III targets was unresolved\",\n        \"Downstream effectors mediating the differentiation phenotype were not identified\"\n      ]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Genome-wide ChIP-seq showing POLR3G- and POLR3GL-containing Pol III occupy identical targets, combined with MYC-selective binding to the POLR3G promoter, established that paralog-specific biology arises from differential transcriptional regulation rather than functional divergence of the subunit proteins.\",\n      \"evidence\": \"ChIP-seq for both Pol III isoforms, MYC ChIP at POLR3G and POLR3GL promoters, quantitative isoform abundance in dividing vs. non-dividing cells\",\n      \"pmids\": [\"24107381\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether subtle target-gene-level quantitative differences exist between isoforms under specific conditions was not excluded\",\n        \"Post-transcriptional regulation of the two subunits was not examined\"\n      ]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Transcriptomic profiling upon POLR3G knockdown in hPSCs revealed that POLR3G influences expression of protein-coding genes, lincRNAs, and snoRNAs beyond canonical Pol III targets, broadening its functional scope in stem cells.\",\n      \"evidence\": \"Deep sequencing of polyA+ and small RNA transcriptomes in POLR3G-depleted hPSCs\",\n      \"pmids\": [\"28494942\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct versus indirect transcriptional effects were not distinguished\",\n        \"Whether effects on splicing and non-coding RNA are mediated through altered tRNA pools or independent mechanisms is unknown\"\n      ]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Studies in Xenopus showed Polr3g and Polr3gL have distinct effects on tRNA isoacceptor pools and that forced Polr3g expression partially reverses myogenic differentiation, extending the differentiation-linked role of POLR3G to a non-mammalian vertebrate.\",\n      \"evidence\": \"Custom tRNA microarray and overexpression in differentiating Xenopus myoblasts\",\n      \"pmids\": [\"31173763\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which altered tRNA isoacceptor ratios influence differentiation state was not established\",\n        \"Relevance of Xenopus findings to mammalian skeletal muscle differentiation not confirmed\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Genetic knockout in mice proved that POLR3G is essential for early embryogenesis while POLR3GL supports later development, and rescue experiments in ESCs demonstrated functional interchangeability of the two subunits when co-expressed, resolving the question of intrinsic versus expression-dependent specificity.\",\n      \"evidence\": \"Constitutive and conditional KO mice, ESC differentiation rescue with exogenous POLR3GL, in vitro transcription and ChIP\",\n      \"pmids\": [\"32576691\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Why POLR3GL cannot compensate in early embryos despite biochemical equivalence (expression timing or level) was not fully dissected\",\n        \"Tissue-specific consequences of POLR3G loss in adult organs remain unexplored\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"CRISPR knockout of POLR3G in triple-negative breast cancer cells demonstrated a direct requirement for anchorage-independent growth, invasion, and in vivo metastasis, linking the Pol III subunit to cancer aggressiveness independently of proliferation rate changes.\",\n      \"evidence\": \"CRISPR KO in MDA-MB231, invasion assays, orthotopic xenograft model, gene expression profiling\",\n      \"pmids\": [\"36497214\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"How POLR3G KO induces FOXA1 and androgen receptor expression mechanistically is unknown\",\n        \"Whether the metastasis phenotype depends on altered Pol III transcriptional output or a non-canonical function was not determined\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Pharmacological rescue experiments placed POLR3G upstream of PI3K/AKT signaling in bladder cancer EMT, providing the first pathway-level mechanism for its pro-invasive effects, while parallel work mapped a gene-internal super-enhancer integrating MYC, ZNF131, ZNF207, and DNA methylation to control POLR3G expression in cancer.\",\n      \"evidence\": \"shRNA/overexpression with LY294002 and 740Y-P pathway modulation in bladder cancer cells; functional TF overexpression plus chromatin architecture analysis\",\n      \"pmids\": [\"37933949\", \"37894362\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Intermediate steps between Pol III transcriptional output and PI3K/AKT activation remain unidentified\",\n        \"Whether the super-enhancer architecture is broadly conserved across cancer types is untested\",\n        \"KDM5B involvement is correlative, not functionally validated\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"RNA-seq upon POLR3G knockdown in bladder cancer implicated Wnt and MAPK pathway components as downstream effectors, expanding the signaling network influenced by POLR3G beyond PI3K/AKT.\",\n      \"evidence\": \"shRNA knockdown, RNA-seq, Western blot validation of Wnt pathway proteins in bladder cancer cells and BBN-induced rat model\",\n      \"pmids\": [\"39039528\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Pathway placement is based on knockdown transcriptomics without epistasis or rescue experiments\",\n        \"Direct versus indirect regulation of Wnt/MAPK components is unresolved\",\n        \"Single-lab study without independent replication\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The mechanism by which altered Pol III transcriptional output (e.g., tRNA pools, other small non-coding RNAs) is transduced into activation of PI3K/AKT, Wnt/MAPK, or other oncogenic signaling pathways remains the central unresolved question.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No direct biochemical link between Pol III transcript changes and signaling pathway activation has been identified\",\n        \"Whether POLR3G has non-canonical, Pol III-independent functions is unexplored\",\n        \"Structural basis of the POLR3G–C34–C82 subcomplex interaction with TFIIIB in mammals lacks atomic-resolution data\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [0, 2, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 1, 2]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [0, 1, 4]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [2, 3, 5]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [7]}\n    ],\n    \"complexes\": [\n      \"RNA polymerase III\",\n      \"C31-C34-C82 subcomplex\"\n    ],\n    \"partners\": [\n      \"POLR3GL\",\n      \"MYC\",\n      \"ZNF131\",\n      \"ZNF207\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}