{"gene":"RPL32","run_date":"2026-06-10T07:46:26","timeline":{"discoveries":[{"year":1997,"finding":"Yeast RPL32 binds to the 5' end of its own pre-mRNA transcript and inhibits splicing; a deletion of a conserved isoleucine residue abolishes this RNA binding and splicing regulation. The same protein also influences pre-rRNA processing in the nucleolus and regulates translation of its own mRNA in the cytoplasm, making it a single protein acting on three distinct RNA substrates in three cellular compartments.","method":"Genetic mutant selection for splicing-regulation deficiency, in vivo labeling, polysome analysis, RNA-binding assays","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (genetic mutants, RNA binding, polysome analysis, rRNA processing assays), replicated across two independent studies (PMID:9121443 and PMID:8366109)","pmids":["9121443","8366109"],"is_preprint":false},{"year":1993,"finding":"Yeast RPL32 regulates translation of its own mRNA through sequences in the 5' leader region; mutations within the 5' leader that abolish splicing regulation also abolish translational regulation, suggesting both are mediated by the same RNA structural element. Excess RPL32 reduces beta-galactosidase production from an L32-leader–LacZ fusion despite increased mRNA levels.","method":"Chimeric gene constructs (L32-leader fused to LacZ), pulse-labeling, polysome fractionation","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal approaches (reporter fusions, pulse labeling, polysome analysis) in a single rigorous study","pmids":["8366109"],"is_preprint":false},{"year":1995,"finding":"The RNA binding target of yeast RPL32 on its own pre-mRNA is a stem–internal loop–stem structural motif of fewer than 30 nucleotides; the internal loop is asymmetric, purine-rich, and closed by a potential G:U pair. Several loop bases are critical for protein binding (Kd ~10 nM) as shown by mutational and chemical protection/modification interference studies.","method":"Chemical and enzymatic RNA probing, thermodynamic melting, mutational analysis, filter-binding assays","journal":"Journal of molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstituted binding with mutagenesis, chemical probing, and thermodynamic characterization; replicated/extended by PMID:8608446 and PMID:9056762","pmids":["7616567"],"is_preprint":false},{"year":1996,"finding":"The G:U pair that closes the internal loop of the RPL32 pre-mRNA binding site is critical for full-strength protein binding; the G residue is required (inosine substitution only modestly reduces binding), while Watson-Crick pairing at that position does not favor binding, indicating the G:U pair influences protein recognition through RNA conformation.","method":"Electrophoretic bandshift and filter-binding assays with 16 sequence variants of a bimolecular stem-loop-stem RNA","journal":"RNA (New York, N.Y.)","confidence":"High","confidence_rationale":"Tier 1 / Moderate — systematic in vitro mutagenesis with two orthogonal binding assays, single lab but comprehensive variant analysis","pmids":["8608446"],"is_preprint":false},{"year":1997,"finding":"In vitro selection (SELEX) of RNA aptamers for yeast RPL32 shows that four purines (two 5'-GA-3' dinucleotides) on both sides of the internal loop are highly conserved and necessary for binding, and that the position but not size of the loop is variable, further defining the protein's RNA recognition requirements.","method":"In vitro RNA selection (SELEX), secondary structure analysis, binding assays","journal":"RNA (New York, N.Y.)","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — rigorous in vitro selection method, single lab, no mutagenesis validation of individual aptamers","pmids":["9056762"],"is_preprint":false},{"year":1989,"finding":"Intron 1 of the mouse rpL32 gene contains a transcriptional regulatory element within its first 27 base pairs that increases expression 5–10-fold; this element functions at the transcriptional level (shown by nuclear run-on) and is position- and orientation-sensitive, distinguishing it from a classical enhancer. Any spliceable intron can fulfill a general role in ensuring efficient RNA yield.","method":"Transfection of deletion/mutant constructs into COS and L cells; nuclear run-on transcription assays","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — deletion mapping combined with nuclear run-on assays, replicated in two cell types with stable and transient transfection","pmids":["2747643"],"is_preprint":false},{"year":1989,"finding":"Maximal transcription of the mouse rpL32 gene requires a ~150–200 bp region spanning the transcriptional start site, including elements at –79 to –69, downstream of the start site in exon I, and in intron 1; distinct nuclear factors bind to these elements including one that also recognizes a motif in the c-myc gene.","method":"Transient-expression assays of chimeric rpL32-CAT genes; gel mobility shift assays","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional deletion mapping combined with gel shift identification of binding factors, single lab with two orthogonal methods","pmids":["2546059"],"is_preprint":false},{"year":1989,"finding":"A downstream element in exon I (containing GGCTGCCATC) is absolutely required for rpL32 transcription in a simple vector context; a nuclear factor specifically binds this sequence as shown by gel mobility shift and methylation interference analysis.","method":"5' deletion and internal deletion mutant transfection into COS/CV-1 cells; gel mobility-shift and methylation interference assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional deletion assay combined with protein-DNA binding characterization, single lab with two orthogonal methods","pmids":["2726762"],"is_preprint":false},{"year":1993,"finding":"The rpL32 promoter contains two binding sites (one in exon I, one in intron 1) for the zinc-finger nuclear protein delta (YY-1/muE1/UCRBP); the two sites function independently and additively to raise expression ~10-fold, and the intronic site functions regardless of orientation. This is a positive role for delta factor, contrasting with its repressive role in other genes.","method":"Transfection of rpL32 genes with site-directed mutations in delta binding sites; gel mobility shift assays","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional mutagenesis combined with binding assays, single lab","pmids":["8341605"],"is_preprint":false},{"year":1993,"finding":"GABP (GA-binding protein), identified by recombinant subunits and specific antibodies, binds a single site (beta element) in the rpL32 promoter forming only dimeric (alpha/beta1 or alpha/beta2) complexes. This solitary site contributes similarly to promoter activity as the proximal site of the tandem rpL32 promoter.","method":"Gel mobility shift with recombinant GABP subunits and GABP-specific antibodies; DNase I footprinting; promoter mutation analysis","journal":"Gene expression","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — recombinant protein binding, antibody supershift, and footprinting in single lab","pmids":["8019128"],"is_preprint":false},{"year":1993,"finding":"Yeast TFIID (TBP) binds directly to the gamma element (~-30 relative to TSS) of the TATA-less rpL32 promoter, and proteins of 20–40 kDa including a 40 kDa species with affinity for canonical TATA elements bind to this element, indicating that rpL32 uses TBP for transcriptional initiation through a non-canonical element.","method":"Gel mobility shift assay with cloned/purified yeast TBP; cell-free transcription competition assays","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — purified recombinant protein binding plus cell-free transcription, single lab","pmids":["8325365"],"is_preprint":false},{"year":1992,"finding":"The 5' terminal oligopyrimidine (5' TOP) sequence of mouse L32 mRNA is required for translational regulation; deletion of this sequence abolishes sequestration of the mRNA in subribosomal (untranslated) particles in quiescent cells. A 56-kDa protein (p56L32) from T-lymphocytes specifically binds the first 34 nucleotides of the L32 5'-UTR including the polypyrimidine tract.","method":"Stable transfection of RSV-L32 constructs with 5'-UTR deletions into 3T3 fibroblasts; polysome fractionation; RNA-protein UV cross-linking/band-shift","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional deletion in transfected cells combined with RNA-protein binding assay, single lab","pmids":["1309750"],"is_preprint":false},{"year":1990,"finding":"RPL32 mRNA redistributes from messenger ribonucleoprotein (mRNP) particles into polysomes following serum or phorbol ester activation of quiescent Swiss 3T3 cells, with the same kinetics as phosphorylation of eIF-4E, consistent with mitogen-induced eIF-4E phosphorylation recruiting translationally controlled mRNAs including L32 mRNA into polysomes.","method":"Polysome gradient fractionation; eIF-4E phosphorylation state analysis by gel electrophoresis; phorbol ester treatment","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — polysome fractionation and parallel biochemical analysis replicated across two mitogenic stimuli, but mechanistic link to eIF-4E remains correlative","pmids":["2303467"],"is_preprint":false},{"year":1995,"finding":"The 56-kDa p56L32 protein from T-lymphocytes requires both the polypyrimidine tract and a downstream element (GGUGGCUGCC) in the L32 5'-UTR for binding; this protein also binds to DNA of identical sequence with similar affinity, suggesting a dual role in transcriptional regulation and translational control.","method":"RNA-protein binding with deletion/site-directed mutants; competition assays with RNA and DNA probes","journal":"European journal of biochemistry","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — mutagenesis-guided binding assays, single lab, no functional consequence of dual binding directly demonstrated","pmids":["7744065"],"is_preprint":false},{"year":1992,"finding":"The beta-region factor (beta element at ~-71 to -70) of the rpL32 promoter is a 55-kDa polypeptide identified by UV cross-linking; a GT→TC mutation at -71/-70 eliminates its binding, and adding excess beta-element oligonucleotide reduces rpL32 transcription in a cell-free system, demonstrating a positive transcriptional role.","method":"UV cross-linking of nuclear extracts to rpL32 promoter fragments; gel mobility shift; cell-free transcription competition assay","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — two orthogonal methods (cross-linking and cell-free transcription), single lab, no protein identification beyond molecular weight","pmids":["1864363"],"is_preprint":false},{"year":1997,"finding":"GABP (the rpL32 beta factor) is constitutively expressed in BC3H1 myoblasts/myocytes; binding of GABP to the rpL32 promoter beta element is reduced in differentiated myocytes and is modulated by phosphorylation (dephosphorylation of extracts increases binding), suggesting post-translational modification of GABP regulates rpL32 transcription during differentiation.","method":"Gel mobility shift assays with recombinant GABP and specific antibodies; dephosphorylation of nuclear extracts; Western blotting for GABP levels","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — recombinant protein, antibody supershift, and phosphorylation manipulation, single lab","pmids":["9138087"],"is_preprint":false},{"year":2000,"finding":"Dexamethasone increases rpL32 gene transcription ~2.5-fold in rat L6 myoblasts and this is accompanied by enhanced binding of the delta factor (but not beta or gamma) to the rpL32 promoter; the glucocorticoid antagonist RU38486 reverses both effects, indicating glucocorticoid-receptor-mediated changes in delta factor activity underlie increased rpL32 transcription.","method":"Nuclear run-on transcription; gel mobility shift assays; pharmacological antagonist (RU38486)","journal":"Molecular and cellular endocrinology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — nuclear run-on combined with binding assays and pharmacological reversal, single lab","pmids":["11000527"],"is_preprint":false},{"year":2020,"finding":"RPL32 knockdown in human lung cancer cells causes ribosomal stress and impaired rRNA maturation; RPL5 and RPL11 then translocate from the nucleus to the nucleoplasm and bind MDM2, preventing MDM2-mediated p53 ubiquitination, leading to p53 accumulation and cell-cycle arrest.","method":"siRNA knockdown; rRNA processing assays; subcellular fractionation; co-immunoprecipitation of RPL5/RPL11 with MDM2; p53 protein level analysis; xenograft model with CpG-RPL32 siRNA","journal":"Molecular therapy. Nucleic acids","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (fractionation, Co-IP, protein levels) in single lab, mechanistic pathway established by KD with specific readouts","pmids":["32516735"],"is_preprint":false},{"year":1977,"finding":"RPL32 (L32) was isolated as a protein of the large (60S) ribosomal subunit of rat liver ribosomes, establishing its physical association with the 60S subunit; its molecular weight and amino acid composition were characterized.","method":"Stepwise LiCl elution from carboxymethylcellulose; ion exchange chromatography; SDS-PAGE; amino acid composition","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct biochemical purification from ribosomes, single study but foundational isolation","pmids":["863909"],"is_preprint":false},{"year":2026,"finding":"Genetic depletion of yeast Rpl32 blocks processing of the initial 35S pre-rRNA, preventing ribosome biogenesis and nuclear export of 60S subunits; this signals to the cytoplasm where mature 18S and 25S rRNAs are degraded in a ribophagy-independent, de-ubiquitination-dependent manner; cyclin 1 mRNA levels rapidly decrease after Rpl32 depletion, and the cell cycle arrests at G1.","method":"Inducible genetic depletion of Rpl32; kinetic analysis of pre-rRNA processing; live-cell imaging of L25-GFP reporter; rRNA degradation assays; mRNA level analysis; cell cycle FACS","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal cellular readouts (rRNA processing, 60S export, rRNA degradation, cell cycle), single lab study","pmids":["42202054"],"is_preprint":false}],"current_model":"RPL32 is an essential component of the 60S ribosomal subunit that, beyond its structural role in translation, autoregulates its own expression via a feedback loop in which the protein binds a purine-rich asymmetric internal-loop RNA structure at the 5' end of its own pre-mRNA to inhibit splicing and suppress translation; it also influences pre-rRNA processing in the nucleolus; its mRNA is subject to 5' TOP-dependent translational control linked to eIF-4E phosphorylation upon mitogenic stimulation; its gene transcription is driven by a complex promoter spanning the cap site that is positively regulated by the delta factor (YY-1), GABP, and other nuclear factors; and in mammalian cells RPL32 knockdown triggers ribosomal stress that causes RPL5/RPL11 to bind and inhibit MDM2, thereby stabilizing p53 and arresting the cell cycle."},"narrative":{"mechanistic_narrative":"RPL32 is an essential structural protein of the large (60S) ribosomal subunit that doubles as an autoregulatory RNA-binding factor coordinating its own expression with ribosome biogenesis [PMID:863909, PMID:9121443, PMID:8366109]. In yeast, a single RPL32 protein acts on three distinct RNA substrates in three compartments: it binds the 5' end of its own pre-mRNA to inhibit splicing, represses translation of its own mRNA through the same 5' leader element, and influences pre-rRNA processing in the nucleolus [PMID:9121443, PMID:8366109]. The autoregulatory binding site is a small stem–internal-loop–stem motif whose asymmetric, purine-rich loop—anchored by conserved 5'-GA dinucleotides and a G:U closing pair—is recognized with nanomolar affinity, with RNA conformation rather than Watson-Crick pairing dictating recognition [PMID:7616567, PMID:8608446, PMID:9056762]. In mammalian cells, RPL32 mRNA carries a 5' terminal oligopyrimidine (TOP) tract that sequesters it in untranslated mRNP particles in quiescent cells and mobilizes it into polysomes upon mitogenic stimulation, coincident with eIF-4E phosphorylation [PMID:1309750, PMID:2303467]. Transcription of the mammalian gene is driven by a complex promoter spanning the cap site, with positive contributions from the delta factor (YY-1), GABP, and TBP acting through exon I and intron 1 elements, and is modulated during differentiation and by glucocorticoids [PMID:2747643, PMID:8341605, PMID:8019128, PMID:8325365, PMID:11000527]. Loss of RPL32 triggers a ribosomal stress response: depletion blocks early pre-rRNA processing and 60S maturation/export, and in human cells frees RPL5 and RPL11 to bind and inhibit MDM2, stabilizing p53 and arresting the cell cycle [PMID:32516735, PMID:42202054].","teleology":[{"year":1977,"claim":"Establishing RPL32 as a physical constituent of the translational machinery defined its baseline identity before any regulatory role was known.","evidence":"Biochemical purification from rat liver 60S ribosomal subunits with SDS-PAGE and amino acid composition analysis","pmids":["863909"],"confidence":"Medium","gaps":["No functional role beyond structural association established","Position within the 60S subunit and rRNA contacts not mapped"]},{"year":1989,"claim":"Dissecting the mammalian rpL32 promoter answered how this ribosomal protein gene achieves high-level transcription, revealing a non-canonical architecture spanning the cap site rather than a classical upstream enhancer.","evidence":"Deletion mapping with CAT/reporter transfections, nuclear run-on, gel mobility shift and methylation interference in COS/L/CV-1 cells","pmids":["2747643","2546059","2726762"],"confidence":"High","gaps":["Identities of several binding factors not resolved","Mechanism by which intron 1 and exon I elements cooperate unclear"]},{"year":1993,"claim":"Identifying the specific factors binding the promoter established which transcriptional regulators drive rpL32, including an unexpected positive role for the normally repressive delta/YY-1 factor.","evidence":"Site-directed promoter mutagenesis with gel shift, footprinting and recombinant-protein/antibody assays for delta (YY-1), GABP, and TBP","pmids":["8341605","8019128","8325365"],"confidence":"Medium","gaps":["How multiple factors integrate into a single transcriptional output not resolved","Beta-element factor identified only by molecular weight in earlier work"]},{"year":1992,"claim":"Defining the 5' TOP element answered how rpL32 mRNA translation is coupled to growth state, showing the TOP tract is required to sequester the message in untranslated particles in quiescent cells.","evidence":"5'-UTR deletion constructs stably transfected into fibroblasts with polysome fractionation; UV cross-linking identifying a 56-kDa binding protein (p56L32)","pmids":["1309750","2303467"],"confidence":"Medium","gaps":["Identity of p56L32 not determined","Link between eIF-4E phosphorylation and L32 mRNA mobilization is correlative, not mechanistic"]},{"year":1995,"claim":"Yeast genetics revealed that RPL32 is itself an RNA-binding autoregulator acting at three levels, and mapped the cis-element required for splicing and translational feedback.","evidence":"Genetic mutant selection, L32-leader–LacZ reporter fusions, polysome analysis, and chemical/enzymatic RNA probing defining a <30-nt stem–internal-loop–stem motif","pmids":["9121443","8366109","7616567"],"confidence":"High","gaps":["Structural basis of protein–RNA contact not solved at atomic resolution","Whether mammalian RPL32 retains the same autoregulatory binding not addressed"]},{"year":1997,"claim":"Systematic mutagenesis and in vitro selection refined the RNA recognition code, showing conserved purines and a conformation-determining G:U pair govern high-affinity binding.","evidence":"Bandshift/filter-binding of sequence variants and SELEX aptamer selection with secondary-structure analysis","pmids":["8608446","9056762"],"confidence":"Medium","gaps":["Individual SELEX aptamers not validated by mutagenesis","Protein residues contacting the conserved purines not identified"]},{"year":2020,"claim":"Linking RPL32 loss to the p53 axis answered what cellular consequence follows disruption of this ribosomal protein, placing it within the ribosomal stress / nucleolar surveillance pathway.","evidence":"siRNA knockdown in human lung cancer cells with rRNA processing assays, subcellular fractionation, RPL5/RPL11–MDM2 co-immunoprecipitation, p53 readouts and xenografts","pmids":["32516735"],"confidence":"Medium","gaps":["Co-IP not reciprocally validated for direct RPL5/RPL11–MDM2 contact","Whether RPL32 itself participates in MDM2 sensing or acts only by triggering stress unclear"]},{"year":2026,"claim":"Inducible yeast depletion dissected the order of events after RPL32 loss, showing the block occurs at the earliest 35S pre-rRNA processing step and propagates to cytoplasmic rRNA degradation and G1 arrest.","evidence":"Inducible genetic depletion with kinetic pre-rRNA processing analysis, L25-GFP export imaging, rRNA degradation assays, cyclin mRNA measurement and cell-cycle FACS","pmids":["42202054"],"confidence":"Medium","gaps":["Mechanism coupling nuclear processing block to cytoplasmic de-ubiquitination-dependent rRNA degradation not defined","Whether the yeast cell-cycle arrest uses a p53-independent route equivalent to the mammalian pathway not addressed"]},{"year":null,"claim":"Whether the yeast three-substrate autoregulatory circuit and the conserved purine-rich RNA recognition mode operate in mammalian RPL32 alongside the 5' TOP/eIF-4E translational control remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No atomic-resolution structure of the RPL32–RNA complex","Cross-species conservation of the splicing-feedback loop untested in mammalian cells","Direct molecular link between eIF-4E phosphorylation and L32 TOP mRNA mobilization unestablished"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003723","term_label":"RNA binding","supporting_discovery_ids":[0,1,2,3,4]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[18]},{"term_id":"GO:0045182","term_label":"translation regulator activity","supporting_discovery_ids":[0,1]}],"localization":[{"term_id":"GO:0005840","term_label":"ribosome","supporting_discovery_ids":[18]},{"term_id":"GO:0005730","term_label":"nucleolus","supporting_discovery_ids":[0,19]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,11]}],"pathway":[{"term_id":"R-HSA-8953854","term_label":"Metabolism of RNA","supporting_discovery_ids":[0,19]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[18,1]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[17,19]}],"complexes":["60S ribosomal subunit"],"partners":["RPL5","RPL11","MDM2","YY1","GABPA","TBP"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P62910","full_name":"Large ribosomal subunit protein eL32","aliases":["60S ribosomal protein L32"],"length_aa":135,"mass_kda":15.9,"function":"Component of the large ribosomal subunit (PubMed:23636399, PubMed:32669547). The ribosome is a large ribonucleoprotein complex responsible for the synthesis of proteins in the cell (PubMed:23636399, PubMed:32669547)","subcellular_location":"Cytoplasm","url":"https://www.uniprot.org/uniprotkb/P62910/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/RPL32","classification":"Common Essential","n_dependent_lines":1206,"n_total_lines":1208,"dependency_fraction":0.9983443708609272},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"EIF2S3","stoichiometry":10.0},{"gene":"EIF3B","stoichiometry":10.0},{"gene":"RBM8A","stoichiometry":10.0},{"gene":"RPL11","stoichiometry":10.0},{"gene":"RPL4","stoichiometry":10.0},{"gene":"RPL5","stoichiometry":10.0},{"gene":"RPS16","stoichiometry":10.0},{"gene":"RPS18","stoichiometry":10.0},{"gene":"CAPRIN1","stoichiometry":4.0},{"gene":"DRG1","stoichiometry":4.0}],"url":"https://opencell.sf.czbiohub.org/search/RPL32","total_profiled":1310},"omim":[],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Endoplasmic reticulum","reliability":"Approved"},{"location":"Cytosol","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/RPL32"},"hgnc":{"alias_symbol":["L32","eL32"],"prev_symbol":[]},"alphafold":{"accession":"P62910","domains":[{"cath_id":"-","chopping":"16-61","consensus_level":"medium","plddt":96.2335,"start":16,"end":61},{"cath_id":"-","chopping":"63-130","consensus_level":"medium","plddt":93.6675,"start":63,"end":130}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P62910","model_url":"https://alphafold.ebi.ac.uk/files/AF-P62910-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P62910-F1-predicted_aligned_error_v6.png","plddt_mean":92.38},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RPL32","jax_strain_url":"https://www.jax.org/strain/search?query=RPL32"},"sequence":{"accession":"P62910","fasta_url":"https://rest.uniprot.org/uniprotkb/P62910.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P62910/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P62910"}},"corpus_meta":[{"pmid":"6327068","id":"PMC_6327068","title":"The gene family encoding the mouse ribosomal protein L32 contains a uniquely expressed intron-containing gene and an unmutated processed gene.","date":"1984","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/6327068","citation_count":376,"is_preprint":false},{"pmid":"2747643","id":"PMC_2747643","title":"Importance of introns for expression of mouse ribosomal protein gene rpL32.","date":"1989","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/2747643","citation_count":134,"is_preprint":false},{"pmid":"9121443","id":"PMC_9121443","title":"Ribosomal protein L32 of Saccharomyces cerevisiae influences both the splicing of its own transcript and the processing of rRNA.","date":"1997","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/9121443","citation_count":118,"is_preprint":false},{"pmid":"2303467","id":"PMC_2303467","title":"Simultaneous cytoplasmic redistribution of ribosomal protein L32 mRNA and phosphorylation of eukaryotic 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translation of its own transcript.","date":"1993","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/8366109","citation_count":78,"is_preprint":false},{"pmid":"25652741","id":"PMC_25652741","title":"Complete plastome sequence of Thalictrum coreanum (Ranunculaceae) and transfer of the rpl32 gene to the nucleus in the ancestor of the subfamily Thalictroideae.","date":"2015","source":"BMC plant biology","url":"https://pubmed.ncbi.nlm.nih.gov/25652741","citation_count":75,"is_preprint":false},{"pmid":"863909","id":"PMC_863909","title":"Isolation of eukaryotic ribosomal proteins. Purification and characterization of 60 S ribosomal subunit proteins L3, L6, L7', L8, L10, L15, L17, L18, L19, L23', L25, L27', L28, L29, L31, L32, L34, L35, L36, L36', and L37'.","date":"1977","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/863909","citation_count":68,"is_preprint":false},{"pmid":"3316213","id":"PMC_3316213","title":"The yeast ribosomal protein L32 and its gene.","date":"1987","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/3316213","citation_count":56,"is_preprint":false},{"pmid":"3866218","id":"PMC_3866218","title":"A processed pseudogene in an intron of the HLA-DP beta 1 chain gene is a member of the ribosomal protein L32 gene family.","date":"1985","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/3866218","citation_count":43,"is_preprint":false},{"pmid":"2726762","id":"PMC_2726762","title":"An element downstream of the cap site is required for transcription of the gene encoding mouse ribosomal protein L32.","date":"1989","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/2726762","citation_count":38,"is_preprint":false},{"pmid":"3437894","id":"PMC_3437894","title":"The synthesis of ribosomal proteins S16 and L32 is not autogenously regulated during mouse myoblast differentiation.","date":"1987","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/3437894","citation_count":36,"is_preprint":false},{"pmid":"8341605","id":"PMC_8341605","title":"The importance of downstream delta-factor binding elements for the activity of the rpL32 promoter.","date":"1993","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/8341605","citation_count":35,"is_preprint":false},{"pmid":"8019128","id":"PMC_8019128","title":"Comparative utilization of transcription factor GABP by the promoters of ribosomal protein genes rpL30 and rpL32.","date":"1993","source":"Gene expression","url":"https://pubmed.ncbi.nlm.nih.gov/8019128","citation_count":31,"is_preprint":false},{"pmid":"7616567","id":"PMC_7616567","title":"Characterization of the pre-mRNA binding site for yeast ribosomal protein L32: the importance of a purine-rich internal loop.","date":"1995","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/7616567","citation_count":31,"is_preprint":false},{"pmid":"33811236","id":"PMC_33811236","title":"The evolutionary fate of rpl32 and rps16 losses in the Euphorbia schimperi (Euphorbiaceae) plastome.","date":"2021","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/33811236","citation_count":25,"is_preprint":false},{"pmid":"32516735","id":"PMC_32516735","title":"RPL32 Promotes Lung Cancer Progression by Facilitating p53 Degradation.","date":"2020","source":"Molecular therapy. Nucleic acids","url":"https://pubmed.ncbi.nlm.nih.gov/32516735","citation_count":24,"is_preprint":false},{"pmid":"22132208","id":"PMC_22132208","title":"Transcriptional downregulation of rice rpL32 gene under abiotic stress is associated with removal of transcription factors within the promoter region.","date":"2011","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22132208","citation_count":19,"is_preprint":false},{"pmid":"7744065","id":"PMC_7744065","title":"Lymphocyte p56L32 is a RNA/DNA-binding protein which interacts with conserved elements of the murine L32 ribosomal protein mRNA.","date":"1995","source":"European journal of biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/7744065","citation_count":18,"is_preprint":false},{"pmid":"2388827","id":"PMC_2388827","title":"Structure of Xenopus laevis ribosomal protein L32 and its expression during development.","date":"1990","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/2388827","citation_count":17,"is_preprint":false},{"pmid":"9138087","id":"PMC_9138087","title":"GA-binding protein is involved in altered expression of ribosomal protein L32 gene.","date":"1997","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9138087","citation_count":17,"is_preprint":false},{"pmid":"8608446","id":"PMC_8608446","title":"Yeast ribosomal protein L32 recognizes an RNA G:U juxtaposition.","date":"1996","source":"RNA (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/8608446","citation_count":16,"is_preprint":false},{"pmid":"9056762","id":"PMC_9056762","title":"RNA apatamers for yeast ribosomal protein L32 have a conserved purine-rich internal loop.","date":"1997","source":"RNA (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/9056762","citation_count":16,"is_preprint":false},{"pmid":"2477362","id":"PMC_2477362","title":"Cloning and analysis of an Escherichia coli operon containing the rpmF gene for ribosomal protein L32 and the gene for a 30-kilodalton protein.","date":"1989","source":"Journal of bacteriology","url":"https://pubmed.ncbi.nlm.nih.gov/2477362","citation_count":14,"is_preprint":false},{"pmid":"14597412","id":"PMC_14597412","title":"Alternative patterns of transcription and translation of the ribosomal protein L32 mRNA in somatic and spermatogenic cells in mice.","date":"2003","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/14597412","citation_count":12,"is_preprint":false},{"pmid":"37569133","id":"PMC_37569133","title":"Sleep-Enhancing Effect of Water Extract from Jujube (Zizyphus jujuba Mill.) Seeds Fermented by Lactobacillus brevis L32.","date":"2023","source":"Foods (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/37569133","citation_count":11,"is_preprint":false},{"pmid":"3911025","id":"PMC_3911025","title":"Gene rpmF for ribosomal protein L32 and gene rimJ for a ribosomal protein acetylating enzyme are located near pyrC (23.4 min) in Escherichia coli.","date":"1985","source":"Molecular & general genetics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/3911025","citation_count":11,"is_preprint":false},{"pmid":"32705264","id":"PMC_32705264","title":"Biological effect of ribosomal protein L32 on human breast cancer cell behavior.","date":"2020","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/32705264","citation_count":10,"is_preprint":false},{"pmid":"1840541","id":"PMC_1840541","title":"Screening a yeast promoter library leads to the isolation of the RP29/L32 and SNR17B/RPL37A divergent promoters and the discovery of a gene encoding 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The same protein also influences pre-rRNA processing in the nucleolus and regulates translation of its own mRNA in the cytoplasm, making it a single protein acting on three distinct RNA substrates in three cellular compartments.\",\n      \"method\": \"Genetic mutant selection for splicing-regulation deficiency, in vivo labeling, polysome analysis, RNA-binding assays\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (genetic mutants, RNA binding, polysome analysis, rRNA processing assays), replicated across two independent studies (PMID:9121443 and PMID:8366109)\",\n      \"pmids\": [\"9121443\", \"8366109\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1993,\n      \"finding\": \"Yeast RPL32 regulates translation of its own mRNA through sequences in the 5' leader region; mutations within the 5' leader that abolish splicing regulation also abolish translational regulation, suggesting both are mediated by the same RNA structural element. Excess RPL32 reduces beta-galactosidase production from an L32-leader–LacZ fusion despite increased mRNA levels.\",\n      \"method\": \"Chimeric gene constructs (L32-leader fused to LacZ), pulse-labeling, polysome fractionation\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal approaches (reporter fusions, pulse labeling, polysome analysis) in a single rigorous study\",\n      \"pmids\": [\"8366109\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"The RNA binding target of yeast RPL32 on its own pre-mRNA is a stem–internal loop–stem structural motif of fewer than 30 nucleotides; the internal loop is asymmetric, purine-rich, and closed by a potential G:U pair. Several loop bases are critical for protein binding (Kd ~10 nM) as shown by mutational and chemical protection/modification interference studies.\",\n      \"method\": \"Chemical and enzymatic RNA probing, thermodynamic melting, mutational analysis, filter-binding assays\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstituted binding with mutagenesis, chemical probing, and thermodynamic characterization; replicated/extended by PMID:8608446 and PMID:9056762\",\n      \"pmids\": [\"7616567\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1996,\n      \"finding\": \"The G:U pair that closes the internal loop of the RPL32 pre-mRNA binding site is critical for full-strength protein binding; the G residue is required (inosine substitution only modestly reduces binding), while Watson-Crick pairing at that position does not favor binding, indicating the G:U pair influences protein recognition through RNA conformation.\",\n      \"method\": \"Electrophoretic bandshift and filter-binding assays with 16 sequence variants of a bimolecular stem-loop-stem RNA\",\n      \"journal\": \"RNA (New York, N.Y.)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — systematic in vitro mutagenesis with two orthogonal binding assays, single lab but comprehensive variant analysis\",\n      \"pmids\": [\"8608446\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"In vitro selection (SELEX) of RNA aptamers for yeast RPL32 shows that four purines (two 5'-GA-3' dinucleotides) on both sides of the internal loop are highly conserved and necessary for binding, and that the position but not size of the loop is variable, further defining the protein's RNA recognition requirements.\",\n      \"method\": \"In vitro RNA selection (SELEX), secondary structure analysis, binding assays\",\n      \"journal\": \"RNA (New York, N.Y.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — rigorous in vitro selection method, single lab, no mutagenesis validation of individual aptamers\",\n      \"pmids\": [\"9056762\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1989,\n      \"finding\": \"Intron 1 of the mouse rpL32 gene contains a transcriptional regulatory element within its first 27 base pairs that increases expression 5–10-fold; this element functions at the transcriptional level (shown by nuclear run-on) and is position- and orientation-sensitive, distinguishing it from a classical enhancer. Any spliceable intron can fulfill a general role in ensuring efficient RNA yield.\",\n      \"method\": \"Transfection of deletion/mutant constructs into COS and L cells; nuclear run-on transcription assays\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — deletion mapping combined with nuclear run-on assays, replicated in two cell types with stable and transient transfection\",\n      \"pmids\": [\"2747643\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1989,\n      \"finding\": \"Maximal transcription of the mouse rpL32 gene requires a ~150–200 bp region spanning the transcriptional start site, including elements at –79 to –69, downstream of the start site in exon I, and in intron 1; distinct nuclear factors bind to these elements including one that also recognizes a motif in the c-myc gene.\",\n      \"method\": \"Transient-expression assays of chimeric rpL32-CAT genes; gel mobility shift assays\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional deletion mapping combined with gel shift identification of binding factors, single lab with two orthogonal methods\",\n      \"pmids\": [\"2546059\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1989,\n      \"finding\": \"A downstream element in exon I (containing GGCTGCCATC) is absolutely required for rpL32 transcription in a simple vector context; a nuclear factor specifically binds this sequence as shown by gel mobility shift and methylation interference analysis.\",\n      \"method\": \"5' deletion and internal deletion mutant transfection into COS/CV-1 cells; gel mobility-shift and methylation interference assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional deletion assay combined with protein-DNA binding characterization, single lab with two orthogonal methods\",\n      \"pmids\": [\"2726762\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1993,\n      \"finding\": \"The rpL32 promoter contains two binding sites (one in exon I, one in intron 1) for the zinc-finger nuclear protein delta (YY-1/muE1/UCRBP); the two sites function independently and additively to raise expression ~10-fold, and the intronic site functions regardless of orientation. This is a positive role for delta factor, contrasting with its repressive role in other genes.\",\n      \"method\": \"Transfection of rpL32 genes with site-directed mutations in delta binding sites; gel mobility shift assays\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional mutagenesis combined with binding assays, single lab\",\n      \"pmids\": [\"8341605\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1993,\n      \"finding\": \"GABP (GA-binding protein), identified by recombinant subunits and specific antibodies, binds a single site (beta element) in the rpL32 promoter forming only dimeric (alpha/beta1 or alpha/beta2) complexes. This solitary site contributes similarly to promoter activity as the proximal site of the tandem rpL32 promoter.\",\n      \"method\": \"Gel mobility shift with recombinant GABP subunits and GABP-specific antibodies; DNase I footprinting; promoter mutation analysis\",\n      \"journal\": \"Gene expression\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — recombinant protein binding, antibody supershift, and footprinting in single lab\",\n      \"pmids\": [\"8019128\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1993,\n      \"finding\": \"Yeast TFIID (TBP) binds directly to the gamma element (~-30 relative to TSS) of the TATA-less rpL32 promoter, and proteins of 20–40 kDa including a 40 kDa species with affinity for canonical TATA elements bind to this element, indicating that rpL32 uses TBP for transcriptional initiation through a non-canonical element.\",\n      \"method\": \"Gel mobility shift assay with cloned/purified yeast TBP; cell-free transcription competition assays\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — purified recombinant protein binding plus cell-free transcription, single lab\",\n      \"pmids\": [\"8325365\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1992,\n      \"finding\": \"The 5' terminal oligopyrimidine (5' TOP) sequence of mouse L32 mRNA is required for translational regulation; deletion of this sequence abolishes sequestration of the mRNA in subribosomal (untranslated) particles in quiescent cells. A 56-kDa protein (p56L32) from T-lymphocytes specifically binds the first 34 nucleotides of the L32 5'-UTR including the polypyrimidine tract.\",\n      \"method\": \"Stable transfection of RSV-L32 constructs with 5'-UTR deletions into 3T3 fibroblasts; polysome fractionation; RNA-protein UV cross-linking/band-shift\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional deletion in transfected cells combined with RNA-protein binding assay, single lab\",\n      \"pmids\": [\"1309750\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1990,\n      \"finding\": \"RPL32 mRNA redistributes from messenger ribonucleoprotein (mRNP) particles into polysomes following serum or phorbol ester activation of quiescent Swiss 3T3 cells, with the same kinetics as phosphorylation of eIF-4E, consistent with mitogen-induced eIF-4E phosphorylation recruiting translationally controlled mRNAs including L32 mRNA into polysomes.\",\n      \"method\": \"Polysome gradient fractionation; eIF-4E phosphorylation state analysis by gel electrophoresis; phorbol ester treatment\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — polysome fractionation and parallel biochemical analysis replicated across two mitogenic stimuli, but mechanistic link to eIF-4E remains correlative\",\n      \"pmids\": [\"2303467\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"The 56-kDa p56L32 protein from T-lymphocytes requires both the polypyrimidine tract and a downstream element (GGUGGCUGCC) in the L32 5'-UTR for binding; this protein also binds to DNA of identical sequence with similar affinity, suggesting a dual role in transcriptional regulation and translational control.\",\n      \"method\": \"RNA-protein binding with deletion/site-directed mutants; competition assays with RNA and DNA probes\",\n      \"journal\": \"European journal of biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — mutagenesis-guided binding assays, single lab, no functional consequence of dual binding directly demonstrated\",\n      \"pmids\": [\"7744065\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1992,\n      \"finding\": \"The beta-region factor (beta element at ~-71 to -70) of the rpL32 promoter is a 55-kDa polypeptide identified by UV cross-linking; a GT→TC mutation at -71/-70 eliminates its binding, and adding excess beta-element oligonucleotide reduces rpL32 transcription in a cell-free system, demonstrating a positive transcriptional role.\",\n      \"method\": \"UV cross-linking of nuclear extracts to rpL32 promoter fragments; gel mobility shift; cell-free transcription competition assay\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — two orthogonal methods (cross-linking and cell-free transcription), single lab, no protein identification beyond molecular weight\",\n      \"pmids\": [\"1864363\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"GABP (the rpL32 beta factor) is constitutively expressed in BC3H1 myoblasts/myocytes; binding of GABP to the rpL32 promoter beta element is reduced in differentiated myocytes and is modulated by phosphorylation (dephosphorylation of extracts increases binding), suggesting post-translational modification of GABP regulates rpL32 transcription during differentiation.\",\n      \"method\": \"Gel mobility shift assays with recombinant GABP and specific antibodies; dephosphorylation of nuclear extracts; Western blotting for GABP levels\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — recombinant protein, antibody supershift, and phosphorylation manipulation, single lab\",\n      \"pmids\": [\"9138087\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"Dexamethasone increases rpL32 gene transcription ~2.5-fold in rat L6 myoblasts and this is accompanied by enhanced binding of the delta factor (but not beta or gamma) to the rpL32 promoter; the glucocorticoid antagonist RU38486 reverses both effects, indicating glucocorticoid-receptor-mediated changes in delta factor activity underlie increased rpL32 transcription.\",\n      \"method\": \"Nuclear run-on transcription; gel mobility shift assays; pharmacological antagonist (RU38486)\",\n      \"journal\": \"Molecular and cellular endocrinology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — nuclear run-on combined with binding assays and pharmacological reversal, single lab\",\n      \"pmids\": [\"11000527\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RPL32 knockdown in human lung cancer cells causes ribosomal stress and impaired rRNA maturation; RPL5 and RPL11 then translocate from the nucleus to the nucleoplasm and bind MDM2, preventing MDM2-mediated p53 ubiquitination, leading to p53 accumulation and cell-cycle arrest.\",\n      \"method\": \"siRNA knockdown; rRNA processing assays; subcellular fractionation; co-immunoprecipitation of RPL5/RPL11 with MDM2; p53 protein level analysis; xenograft model with CpG-RPL32 siRNA\",\n      \"journal\": \"Molecular therapy. Nucleic acids\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (fractionation, Co-IP, protein levels) in single lab, mechanistic pathway established by KD with specific readouts\",\n      \"pmids\": [\"32516735\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1977,\n      \"finding\": \"RPL32 (L32) was isolated as a protein of the large (60S) ribosomal subunit of rat liver ribosomes, establishing its physical association with the 60S subunit; its molecular weight and amino acid composition were characterized.\",\n      \"method\": \"Stepwise LiCl elution from carboxymethylcellulose; ion exchange chromatography; SDS-PAGE; amino acid composition\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct biochemical purification from ribosomes, single study but foundational isolation\",\n      \"pmids\": [\"863909\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Genetic depletion of yeast Rpl32 blocks processing of the initial 35S pre-rRNA, preventing ribosome biogenesis and nuclear export of 60S subunits; this signals to the cytoplasm where mature 18S and 25S rRNAs are degraded in a ribophagy-independent, de-ubiquitination-dependent manner; cyclin 1 mRNA levels rapidly decrease after Rpl32 depletion, and the cell cycle arrests at G1.\",\n      \"method\": \"Inducible genetic depletion of Rpl32; kinetic analysis of pre-rRNA processing; live-cell imaging of L25-GFP reporter; rRNA degradation assays; mRNA level analysis; cell cycle FACS\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal cellular readouts (rRNA processing, 60S export, rRNA degradation, cell cycle), single lab study\",\n      \"pmids\": [\"42202054\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RPL32 is an essential component of the 60S ribosomal subunit that, beyond its structural role in translation, autoregulates its own expression via a feedback loop in which the protein binds a purine-rich asymmetric internal-loop RNA structure at the 5' end of its own pre-mRNA to inhibit splicing and suppress translation; it also influences pre-rRNA processing in the nucleolus; its mRNA is subject to 5' TOP-dependent translational control linked to eIF-4E phosphorylation upon mitogenic stimulation; its gene transcription is driven by a complex promoter spanning the cap site that is positively regulated by the delta factor (YY-1), GABP, and other nuclear factors; and in mammalian cells RPL32 knockdown triggers ribosomal stress that causes RPL5/RPL11 to bind and inhibit MDM2, thereby stabilizing p53 and arresting the cell cycle.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RPL32 is an essential structural protein of the large (60S) ribosomal subunit that doubles as an autoregulatory RNA-binding factor coordinating its own expression with ribosome biogenesis [#18, #0]. In yeast, a single RPL32 protein acts on three distinct RNA substrates in three compartments: it binds the 5' end of its own pre-mRNA to inhibit splicing, represses translation of its own mRNA through the same 5' leader element, and influences pre-rRNA processing in the nucleolus [#0, #1]. The autoregulatory binding site is a small stem–internal-loop–stem motif whose asymmetric, purine-rich loop—anchored by conserved 5'-GA dinucleotides and a G:U closing pair—is recognized with nanomolar affinity, with RNA conformation rather than Watson-Crick pairing dictating recognition [#2, #3, #4]. In mammalian cells, RPL32 mRNA carries a 5' terminal oligopyrimidine (TOP) tract that sequesters it in untranslated mRNP particles in quiescent cells and mobilizes it into polysomes upon mitogenic stimulation, coincident with eIF-4E phosphorylation [#11, #12]. Transcription of the mammalian gene is driven by a complex promoter spanning the cap site, with positive contributions from the delta factor (YY-1), GABP, and TBP acting through exon I and intron 1 elements, and is modulated during differentiation and by glucocorticoids [#5, #8, #9, #10, #16]. Loss of RPL32 triggers a ribosomal stress response: depletion blocks early pre-rRNA processing and 60S maturation/export, and in human cells frees RPL5 and RPL11 to bind and inhibit MDM2, stabilizing p53 and arresting the cell cycle [#17, #19].\",\n  \"teleology\": [\n    {\n      \"year\": 1977,\n      \"claim\": \"Establishing RPL32 as a physical constituent of the translational machinery defined its baseline identity before any regulatory role was known.\",\n      \"evidence\": \"Biochemical purification from rat liver 60S ribosomal subunits with SDS-PAGE and amino acid composition analysis\",\n      \"pmids\": [\"863909\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No functional role beyond structural association established\", \"Position within the 60S subunit and rRNA contacts not mapped\"]\n    },\n    {\n      \"year\": 1989,\n      \"claim\": \"Dissecting the mammalian rpL32 promoter answered how this ribosomal protein gene achieves high-level transcription, revealing a non-canonical architecture spanning the cap site rather than a classical upstream enhancer.\",\n      \"evidence\": \"Deletion mapping with CAT/reporter transfections, nuclear run-on, gel mobility shift and methylation interference in COS/L/CV-1 cells\",\n      \"pmids\": [\"2747643\", \"2546059\", \"2726762\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identities of several binding factors not resolved\", \"Mechanism by which intron 1 and exon I elements cooperate unclear\"]\n    },\n    {\n      \"year\": 1993,\n      \"claim\": \"Identifying the specific factors binding the promoter established which transcriptional regulators drive rpL32, including an unexpected positive role for the normally repressive delta/YY-1 factor.\",\n      \"evidence\": \"Site-directed promoter mutagenesis with gel shift, footprinting and recombinant-protein/antibody assays for delta (YY-1), GABP, and TBP\",\n      \"pmids\": [\"8341605\", \"8019128\", \"8325365\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How multiple factors integrate into a single transcriptional output not resolved\", \"Beta-element factor identified only by molecular weight in earlier work\"]\n    },\n    {\n      \"year\": 1992,\n      \"claim\": \"Defining the 5' TOP element answered how rpL32 mRNA translation is coupled to growth state, showing the TOP tract is required to sequester the message in untranslated particles in quiescent cells.\",\n      \"evidence\": \"5'-UTR deletion constructs stably transfected into fibroblasts with polysome fractionation; UV cross-linking identifying a 56-kDa binding protein (p56L32)\",\n      \"pmids\": [\"1309750\", \"2303467\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Identity of p56L32 not determined\", \"Link between eIF-4E phosphorylation and L32 mRNA mobilization is correlative, not mechanistic\"]\n    },\n    {\n      \"year\": 1995,\n      \"claim\": \"Yeast genetics revealed that RPL32 is itself an RNA-binding autoregulator acting at three levels, and mapped the cis-element required for splicing and translational feedback.\",\n      \"evidence\": \"Genetic mutant selection, L32-leader–LacZ reporter fusions, polysome analysis, and chemical/enzymatic RNA probing defining a <30-nt stem–internal-loop–stem motif\",\n      \"pmids\": [\"9121443\", \"8366109\", \"7616567\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of protein–RNA contact not solved at atomic resolution\", \"Whether mammalian RPL32 retains the same autoregulatory binding not addressed\"]\n    },\n    {\n      \"year\": 1997,\n      \"claim\": \"Systematic mutagenesis and in vitro selection refined the RNA recognition code, showing conserved purines and a conformation-determining G:U pair govern high-affinity binding.\",\n      \"evidence\": \"Bandshift/filter-binding of sequence variants and SELEX aptamer selection with secondary-structure analysis\",\n      \"pmids\": [\"8608446\", \"9056762\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Individual SELEX aptamers not validated by mutagenesis\", \"Protein residues contacting the conserved purines not identified\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Linking RPL32 loss to the p53 axis answered what cellular consequence follows disruption of this ribosomal protein, placing it within the ribosomal stress / nucleolar surveillance pathway.\",\n      \"evidence\": \"siRNA knockdown in human lung cancer cells with rRNA processing assays, subcellular fractionation, RPL5/RPL11–MDM2 co-immunoprecipitation, p53 readouts and xenografts\",\n      \"pmids\": [\"32516735\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Co-IP not reciprocally validated for direct RPL5/RPL11–MDM2 contact\", \"Whether RPL32 itself participates in MDM2 sensing or acts only by triggering stress unclear\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Inducible yeast depletion dissected the order of events after RPL32 loss, showing the block occurs at the earliest 35S pre-rRNA processing step and propagates to cytoplasmic rRNA degradation and G1 arrest.\",\n      \"evidence\": \"Inducible genetic depletion with kinetic pre-rRNA processing analysis, L25-GFP export imaging, rRNA degradation assays, cyclin mRNA measurement and cell-cycle FACS\",\n      \"pmids\": [\"42202054\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism coupling nuclear processing block to cytoplasmic de-ubiquitination-dependent rRNA degradation not defined\", \"Whether the yeast cell-cycle arrest uses a p53-independent route equivalent to the mammalian pathway not addressed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Whether the yeast three-substrate autoregulatory circuit and the conserved purine-rich RNA recognition mode operate in mammalian RPL32 alongside the 5' TOP/eIF-4E translational control remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No atomic-resolution structure of the RPL32–RNA complex\", \"Cross-species conservation of the splicing-feedback loop untested in mammalian cells\", \"Direct molecular link between eIF-4E phosphorylation and L32 TOP mRNA mobilization unestablished\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003723\", \"supporting_discovery_ids\": [0, 1, 2, 3, 4]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [18]},\n      {\"term_id\": \"GO:0045182\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005840\", \"supporting_discovery_ids\": [18]},\n      {\"term_id\": \"GO:0005730\", \"supporting_discovery_ids\": [0, 19]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 11]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-8953854\", \"supporting_discovery_ids\": [0, 19]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [18, 1]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [17, 19]}\n    ],\n    \"complexes\": [\"60S ribosomal subunit\"],\n    \"partners\": [\"RPL5\", \"RPL11\", \"MDM2\", \"YY1\", \"GABPA\", \"TBP\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}