{"gene":"RFC2","run_date":"2026-06-10T06:43:36","timeline":{"discoveries":[{"year":2001,"finding":"The conserved lysine (K71) in the Walker A motif (ATP-binding domain) of yeast Rfc2 is essential for ATPase activity, DNA binding, and clamp loading. The rfc2-K71E mutation severely impairs ATPase, clamp loading, and DNA binding; the conservative rfc2-K71R mutation has milder defects suppressible at high ATP concentrations. All mutant RFC complexes retain PCNA interaction.","method":"Active-site mutagenesis (Walker A lysine → glutamate or arginine), in vitro ATPase assay, clamp-loading assay, DNA-binding assay, and PCNA-interaction assay with bacterially overproduced mutant RFC complexes","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with active-site mutagenesis and multiple orthogonal functional assays in a single rigorous study","pmids":["11432854"],"is_preprint":false},{"year":1994,"finding":"Yeast Rfc2 protein is a component of the RF-C complex required for DNA replication: it co-purifies with RF-C activity, and polyclonal antibodies against bacterially expressed Rfc2 specifically reduce RF-C activity in a DNA polymerase III-dependent replication reaction. Bacterially expressed Rfc2 preferentially binds primed single-stranded DNA and weakly binds ATP.","method":"Co-purification of Rfc2 with RF-C activity, antibody inhibition assay, in vitro DNA/ATP binding assay with recombinant protein, RFC2 gene disruption (lethal, dumbbell morphology)","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — multiple orthogonal methods (co-purification, antibody inhibition, in vitro binding) in a single study establishing Rfc2 as an RFC subunit","pmids":["8202350"],"is_preprint":false},{"year":1998,"finding":"Yeast RFC2 is required for chromosomal DNA replication and an S-phase DNA damage checkpoint. The rfc2-1 thermosensitive mutation causes DNA integrity defects, sensitivity to hydroxyurea/MMS/UV, elevated mitotic recombination/chromosome loss, and checkpoint failure. Genetic epistasis shows synthetic lethality with cdc44-1/rfc1 and rfc5-1 mutations, exacerbation by cdc2-2 and pol2-11 mutations, and suppression by multicopy RFC5, indicating functional interaction with Rfc1, Rfc5, and DNA polymerases δ and ε.","method":"Thermosensitive allele (rfc2-1) genetic analysis; DNA damage sensitivity assays; mitotic recombination and chromosome loss assays; synthetic lethality and multicopy suppression screens","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal genetic methods (epistasis, suppression, synthetic lethality) in yeast with defined phenotypic readouts, replicated across multiple assays","pmids":["9671499"],"is_preprint":false},{"year":2008,"finding":"Human RFC2 is monoubiquitylated by the RAD6-RAD18 E2-E3 ubiquitin ligase complex in response to DNA-damaging agents (alkylating agents, H2O2) and this ubiquitylation is inhibited by RPA (replication protein A). A D228A mutation in RFC2 (corresponding to a yeast Rfc4 mutation that reduces RPA interaction) causes constitutive RFC2 ubiquitylation even without DNA damage, establishing that RPA binding to RFC2 suppresses its ubiquitylation. RAD6-RAD18-mediated ubiquitylation of RFC2 was reconstituted in vitro.","method":"In vitro ubiquitylation assay (RAD6-RAD18 complex + RFC2), site-directed mutagenesis (D228A), in-cell ubiquitylation assays with RAD18-dependent damage response, RPA inhibition assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution of ubiquitylation with mutagenesis and RPA inhibition, supported by in-cell validation; single lab but multiple orthogonal methods","pmids":["18245774"],"is_preprint":false},{"year":2005,"finding":"RFC40 (RFC2) directly interacts with the regulatory subunit RIα of cAMP-dependent PKA. The interaction maps to the N-terminus of RIα and the C-terminus of RFC40. RFC37 (RFC3) competes with RIα and displaces it from the RFC40-RIα complex. RIα functions as a nuclear transport protein for RFC40, and impairment of this nuclear transport arrests cells in G1 phase.","method":"Yeast two-hybrid screening (RFC40 as bait), co-immunoprecipitation, interaction domain mapping, cell cycle analysis (flow cytometry) after nuclear transport disruption","journal":"Cancer biology & therapy / Cell cycle (Georgetown, Tex.)","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — yeast two-hybrid plus Co-IP for interaction, domain mapping, and functional cell cycle readout; two papers from same lab using overlapping methods","pmids":["15655353","15846072"],"is_preprint":false},{"year":2006,"finding":"CDK2/Cyclin E phosphorylates RIα at a serine residue, promoting dissociation of the RIα-RFC40 complex. In vitro phosphorylation of RIα by CDK2/CyclinE prevents its association with RFC40. CDK inhibitor olomoucine increases the RIα-RFC40 complex and decreases the RFC40-RFC37 complex. Inhibition of phosphatase PP1 by Calyculin A reduces RIα-RFC40 complex formation, consistent with phosphorylation driving dissociation.","method":"In vitro kinase assay (CDK2/CyclinE + RIα), CDK inhibitor treatment (olomoucine), phosphatase inhibitor treatment (Calyculin A), co-immunoprecipitation of complex formation","journal":"Cell cycle (Georgetown, Tex.)","confidence":"Medium","confidence_rationale":"Tier 1-2 / Weak — in vitro phosphorylation assay with pharmacological inhibitors and Co-IP; single lab, multiple methods but no direct mutagenesis of the phosphosite","pmids":["16582606"],"is_preprint":false},{"year":2009,"finding":"In fission yeast, pentapeptide insertions in Rfc2 that abolish growth cluster near the ATP-binding sites (arginine finger motif and P-loop/sites C and D) and the central five-stranded β-sheet, as mapped onto the 3D structure of budding yeast Rfc2. Non-lethal insertions map predominantly to loop regions or outer surface of the RFC complex.","method":"Pentapeptide-scanning mutagenesis of fission yeast rfc2; growth complementation assay in rfc2Δ cells; structural mapping onto 3D structure of budding yeast Rfc2","journal":"The FEBS journal","confidence":"Medium","confidence_rationale":"Tier 1-2 / Weak — systematic mutagenesis with structural mapping; single lab, in vivo growth assay only (no in vitro biochemical activity measurements)","pmids":["19664060"],"is_preprint":false},{"year":2006,"finding":"cAMP (via N6-monobutyryl cAMP) upregulates RFC40 mRNA and protein expression, increases RIα-RFC40 complex formation, increases the nuclear-to-cytoplasmic ratio of RFC40, and increases the proportion of S-phase cells, while decreasing RFC40-RFC37 complex formation and DNA replication rate.","method":"cAMP analog treatment of MCF7 cells; qRT-PCR; western blot; co-immunoprecipitation; nuclear/cytoplasmic fractionation; flow cytometry cell cycle analysis","journal":"Experimental cell research","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pharmacological treatment with indirect readouts; no direct mechanistic dissection of how cAMP controls RFC40","pmids":["16413017"],"is_preprint":false},{"year":2020,"finding":"In glioblastoma cells, NELFA mRNA (noncoding mode) interacts with Rad17 and regulates the interaction between Rad17 and the RFC2-5 complex, with downstream impact on CHK1, CHK2, and BRCA1 phosphorylation in DNA damage repair signaling.","method":"Co-immunoprecipitation, RNA immunoprecipitation, knockdown experiments measuring CHK1/CHK2/BRCA1 phosphorylation","journal":"Molecular oncology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab Co-IP; RFC2 participates as part of a multi-subunit complex but its specific role is not mechanistically dissected; indirect evidence","pmids":["31845510"],"is_preprint":false},{"year":2012,"finding":"Knockdown of RFC40 (RFC2) in neonatal rat cardiac myocytes causes chromosomal missegregation/aneuploidy and decrease in cell numbers, establishing a direct role for RFC40 in maintaining chromosomal integrity during cardiac myocyte replication.","method":"siRNA knockdown of RFC40 in neonatal rat cardiac myocytes; FISH for chromosome 12 to detect missegregation/aneuploidy; cell counting","journal":"PloS one","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, siRNA knockdown with FISH readout; no mechanistic pathway dissection beyond the phenotypic result","pmids":["22720015"],"is_preprint":false},{"year":2024,"finding":"rfc2 knockout zebrafish (CRISPR-Cas9) exhibit small head/brain, jaw/dental defects, and vascular problems reminiscent of Williams syndrome. RNA-seq reveals that genes associated with neural cell survival and differentiation are specifically affected. Heterozygous rfc2 KO adult zebrafish show anxiety-like behavior with increased social cohesion.","method":"CRISPR-Cas9 knockout of rfc2 in zebrafish; phenotypic analysis; RNA-seq transcriptome analysis; behavioral assay","journal":"Journal of genetics and genomics","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — in vivo loss-of-function with defined phenotypic readouts and transcriptomic analysis; single lab, novel organism model without biochemical mechanism","pmids":["39368701"],"is_preprint":false}],"current_model":"RFC2 (RFC40) is the second-largest subunit of the heteropentameric Replication Factor C (RFC) complex; its Walker A ATP-binding domain (K71 in yeast) is essential for ATPase activity, DNA binding, and PCNA clamp loading onto primed DNA templates, while its arginine finger motif and central β-sheet are also structurally critical; it is subject to RAD6-RAD18-dependent monoubiquitylation after DNA damage, a modification suppressed by RPA binding to RFC2; in human cells its nuclear import depends on interaction with PKA regulatory subunit RIα (mapping to the RFC40 C-terminus), a complex whose dissociation is triggered by CDK2/CyclinE phosphorylation of RIα and is required for G1-to-S phase progression; genetic studies in yeast establish functional interactions with Rfc1, Rfc5, and DNA polymerases δ and ε, and an S-phase checkpoint role."},"narrative":{"mechanistic_narrative":"RFC2 (RFC40) is an essential subunit of the heteropentameric Replication Factor C (RFC) clamp-loader complex that loads the PCNA sliding clamp onto primed DNA during chromosomal replication and supports the S-phase DNA damage checkpoint [PMID:8202350, PMID:9671499]. Its conserved Walker A lysine (K71 in yeast) is required for ATPase activity, primed-template DNA binding, and clamp loading, while the arginine finger motif and central β-sheet are structurally critical for function [PMID:11432854, PMID:19664060]. RFC2 acts within a network of replication factors, showing functional interactions with Rfc1, Rfc5, and DNA polymerases δ and ε [PMID:9671499]. The protein is regulated by post-translational and protein-interaction controls: it is monoubiquitylated by the RAD6-RAD18 E2-E3 ligase complex following DNA damage, a modification suppressed by RPA binding to RFC2 [PMID:18245774], and in human cells its nuclear import depends on interaction with the PKA regulatory subunit RIα, a complex displaced by RFC3 (RFC37) and dissolved upon CDK2/Cyclin E phosphorylation of RIα to permit G1-to-S progression [PMID:15655353, PMID:15846072, PMID:16582606]. Loss of RFC2 function causes chromosomal missegregation and aneuploidy, and rfc2 knockout in zebrafish produces craniofacial, brain, and vascular defects with altered neural survival/differentiation gene expression [PMID:22720015, PMID:39368701].","teleology":[{"year":1994,"claim":"Established that Rfc2 is a bona fide subunit of the RF-C clamp-loader complex required for DNA replication, defining its place in the replication machinery.","evidence":"Co-purification with RF-C activity, antibody inhibition of a polymerase III-dependent replication reaction, in vitro DNA/ATP binding, and lethal gene disruption in yeast","pmids":["8202350"],"confidence":"High","gaps":["Did not resolve the stoichiometry or catalytic mechanism within the complex","ATP binding characterized only as weak with recombinant protein"]},{"year":1998,"claim":"Connected RFC2 not only to bulk replication but to an S-phase DNA damage checkpoint and placed it genetically with Rfc1, Rfc5, and polymerases δ/ε.","evidence":"Thermosensitive rfc2-1 allele with DNA damage sensitivity, recombination/chromosome loss assays, synthetic lethality and multicopy suppression in yeast","pmids":["9671499"],"confidence":"High","gaps":["Genetic interactions do not establish direct physical contacts","Molecular basis of the checkpoint defect not dissected"]},{"year":2001,"claim":"Pinpointed the Walker A lysine K71 as essential for the ATPase, DNA-binding, and clamp-loading activities, separating catalytic function from PCNA association.","evidence":"Active-site mutagenesis (K71E/K71R) with in vitro ATPase, clamp-loading, DNA-binding, and PCNA-interaction assays on reconstituted RFC complexes","pmids":["11432854"],"confidence":"High","gaps":["Did not map other catalytic residues to function","Mechanism of ATP-driven clamp opening not resolved"]},{"year":2005,"claim":"Identified a non-replication regulatory axis: RFC40 binds PKA regulatory subunit RIα, which serves as its nuclear transport carrier, linking RFC40 nuclear import to cell cycle progression.","evidence":"Yeast two-hybrid, Co-IP, domain mapping (RIα N-terminus to RFC40 C-terminus), RFC3 competition, and G1-arrest cell cycle analysis","pmids":["15655353","15846072"],"confidence":"Medium","gaps":["Whether RIα transports free RFC40 or assembled RFC unclear","Single lab with overlapping methods"]},{"year":2006,"claim":"Showed CDK2/Cyclin E phosphorylation of RIα dissolves the RIα-RFC40 complex, providing a cell-cycle switch governing RFC40 availability.","evidence":"In vitro kinase assay, CDK inhibitor (olomoucine) and phosphatase inhibitor (Calyculin A) treatments, Co-IP of complex changes","pmids":["16582606"],"confidence":"Medium","gaps":["Phosphosite not confirmed by mutagenesis","Single lab, pharmacological inference"]},{"year":2008,"claim":"Defined a DNA-damage-responsive modification of RFC2: RAD6-RAD18 monoubiquitylation antagonized by RPA binding, coupling RFC2 to damage signaling.","evidence":"In vitro reconstituted ubiquitylation, D228A mutant causing constitutive ubiquitylation, in-cell RAD18-dependent damage response, RPA inhibition assay","pmids":["18245774"],"confidence":"High","gaps":["Functional consequence of the ubiquitin mark on clamp loading not established","Ubiquitylation site(s) on RFC2 not mapped"]},{"year":2009,"claim":"Mapped which structural elements are indispensable, confirming ATP-binding sites and the central β-sheet as functionally critical surfaces.","evidence":"Pentapeptide-scanning mutagenesis of fission yeast rfc2 with growth complementation and structural mapping onto budding yeast Rfc2","pmids":["19664060"],"confidence":"Medium","gaps":["In vivo growth only; no in vitro biochemical activity measurements","Effects of individual insertions on clamp loading not measured"]},{"year":2012,"claim":"Demonstrated a direct cellular requirement for RFC40 in chromosomal integrity, with depletion causing missegregation and aneuploidy.","evidence":"siRNA knockdown in neonatal rat cardiac myocytes with chromosome-12 FISH and cell counting","pmids":["22720015"],"confidence":"Low","gaps":["Single lab phenotypic study without mechanistic dissection","Off-target effects of siRNA not controlled by rescue"]},{"year":2020,"claim":"Placed the RFC2-5 complex within damage-response signaling via Rad17, regulated by NELFA RNA, affecting CHK1/CHK2/BRCA1 phosphorylation.","evidence":"Co-IP, RNA immunoprecipitation, knockdown measuring checkpoint kinase phosphorylation in glioblastoma cells","pmids":["31845510"],"confidence":"Low","gaps":["RFC2-specific contribution within the complex not isolated","Indirect, single-lab evidence"]},{"year":2024,"claim":"Established an organismal developmental requirement, with rfc2 knockout causing craniofacial, neural, and vascular defects resembling a human syndrome.","evidence":"CRISPR-Cas9 rfc2 knockout zebrafish, phenotyping, RNA-seq, and behavioral assays","pmids":["39368701"],"confidence":"Medium","gaps":["No biochemical mechanism linking RFC2 loss to phenotypes","Causal disease relationship in humans not established by this model"]},{"year":null,"claim":"How DNA-damage monoubiquitylation, RIα/PKA-dependent trafficking, and clamp-loading activity are integrated into a single regulatory program for RFC2 across cell cycle and development remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structure of human RFC2 within an assembled clamp-loader in the corpus","Functional consequence of RFC2 monoubiquitylation undefined","Connection between developmental phenotypes and replication function unestablished"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[0]},{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[0,1]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[4,7]}],"pathway":[{"term_id":"R-HSA-69306","term_label":"DNA Replication","supporting_discovery_ids":[1,2]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[3,8]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[4,5]}],"complexes":["Replication Factor C (RFC) complex"],"partners":["RFC1","RFC5","RFC3","PCNA","RAD18","RAD6","RPA","PRKAR1A"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P35250","full_name":"Replication factor C subunit 2","aliases":["Activator 1 40 kDa subunit","A1 40 kDa subunit","Activator 1 subunit 2","Replication factor C 40 kDa subunit","RF-C 40 kDa subunit","RFC40"],"length_aa":354,"mass_kda":39.2,"function":"Subunit of the replication factor C (RFC) complex which acts during elongation of primed DNA templates by DNA polymerases delta and epsilon, and is necessary for ATP-dependent loading of proliferating cell nuclear antigen (PCNA) onto primed DNA (PubMed:9488738). This subunit binds ATP (By similarity)","subcellular_location":"Nucleus","url":"https://www.uniprot.org/uniprotkb/P35250/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/RFC2","classification":"Common Essential","n_dependent_lines":1202,"n_total_lines":1208,"dependency_fraction":0.9950331125827815},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"HIST2H2BE","stoichiometry":0.2},{"gene":"HMGA1","stoichiometry":0.2},{"gene":"HUS1","stoichiometry":0.2},{"gene":"NPM1","stoichiometry":0.2},{"gene":"PARP1","stoichiometry":0.2},{"gene":"PCNA","stoichiometry":0.2},{"gene":"RAD17","stoichiometry":0.2},{"gene":"RPS16","stoichiometry":0.2},{"gene":"SSRP1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/RFC2","total_profiled":1310},"omim":[{"mim_id":"616714","title":"HMG-BOX TRANSCRIPTION FACTOR 1; HBP1","url":"https://www.omim.org/entry/616714"},{"mim_id":"613203","title":"DNA REPLICATION AND SISTER CHROMATID COHESION 1; DSCC1","url":"https://www.omim.org/entry/613203"},{"mim_id":"613202","title":"CHROMOSOME TRANSMISSION FIDELITY FACTOR 8; CHTF8","url":"https://www.omim.org/entry/613202"},{"mim_id":"613201","title":"CHROMOSOME TRANSMISSION FIDELITY FACTOR 18; CHTF18","url":"https://www.omim.org/entry/613201"},{"mim_id":"609309","title":"MutS HOMOLOG 2; MSH2","url":"https://www.omim.org/entry/609309"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Golgi apparatus","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/RFC2"},"hgnc":{"alias_symbol":["A1","RFC40"],"prev_symbol":[]},"alphafold":{"accession":"P35250","domains":[{"cath_id":"3.40.50.300","chopping":"40-190","consensus_level":"high","plddt":90.2446,"start":40,"end":190},{"cath_id":"1.10.8.60","chopping":"196-256","consensus_level":"high","plddt":93.6577,"start":196,"end":256},{"cath_id":"1.20.272.10","chopping":"261-349","consensus_level":"high","plddt":92.7298,"start":261,"end":349}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P35250","model_url":"https://alphafold.ebi.ac.uk/files/AF-P35250-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P35250-F1-predicted_aligned_error_v6.png","plddt_mean":86.69},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RFC2","jax_strain_url":"https://www.jax.org/strain/search?query=RFC2"},"sequence":{"accession":"P35250","fasta_url":"https://rest.uniprot.org/uniprotkb/P35250.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P35250/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P35250"}},"corpus_meta":[{"pmid":"11432854","id":"PMC_11432854","title":"ATP utilization by yeast replication factor C. III. The ATP-binding domains of Rfc2, Rfc3, and Rfc4 are essential for DNA recognition and clamp loading.","date":"2001","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11432854","citation_count":58,"is_preprint":false},{"pmid":"9671499","id":"PMC_9671499","title":"The RFC2 gene, encoding the third-largest subunit of the replication factor C complex, is required for an S-phase checkpoint in Saccharomyces cerevisiae.","date":"1998","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/9671499","citation_count":57,"is_preprint":false},{"pmid":"7774928","id":"PMC_7774928","title":"Assignment of the 36.5-kDa (RFC5), 37-kDa (RFC4), 38-kDa (RFC3), and 40-kDa (RFC2) subunit genes of human replication factor C to chromosome bands 12q24.2-q24.3, 3q27, 13q12.3-q13, and 7q11.23.","date":"1995","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/7774928","citation_count":40,"is_preprint":false},{"pmid":"8202350","id":"PMC_8202350","title":"The RFC2 gene encoding a subunit of replication factor C of Saccharomyces cerevisiae.","date":"1994","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/8202350","citation_count":38,"is_preprint":false},{"pmid":"18245774","id":"PMC_18245774","title":"DNA damage-induced ubiquitylation of RFC2 subunit of replication factor C complex.","date":"2008","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/18245774","citation_count":33,"is_preprint":false},{"pmid":"15846072","id":"PMC_15846072","title":"RIalpha influences cellular proliferation in cancer cells by transporting RFC40 into the nucleus.","date":"2005","source":"Cancer biology & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/15846072","citation_count":33,"is_preprint":false},{"pmid":"32239691","id":"PMC_32239691","title":"RFC2, a direct target of miR-744, modulates the cell cycle and promotes the proliferation of CRC cells.","date":"2020","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/32239691","citation_count":26,"is_preprint":false},{"pmid":"11003705","id":"PMC_11003705","title":"Comparative genomic sequence analysis of the Williams syndrome region (LIMK1-RFC2) of human chromosome 7q11.23.","date":"2000","source":"Mammalian genome : official journal of the International Mammalian Genome Society","url":"https://pubmed.ncbi.nlm.nih.gov/11003705","citation_count":25,"is_preprint":false},{"pmid":"32422304","id":"PMC_32422304","title":"Xanthohumol regulates miR-4749-5p-inhibited RFC2 signaling in enhancing temozolomide cytotoxicity to glioblastoma.","date":"2020","source":"Life 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gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/37821801","citation_count":6,"is_preprint":false},{"pmid":"16413017","id":"PMC_16413017","title":"Cyclic AMP regulates the expression and nuclear translocation of RFC40 in MCF7 cells.","date":"2006","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/16413017","citation_count":4,"is_preprint":false},{"pmid":"31845510","id":"PMC_31845510","title":"The noncoding function of NELFA mRNA promotes the development of oesophageal squamous cell carcinoma by regulating the Rad17-RFC2-5 complex.","date":"2020","source":"Molecular oncology","url":"https://pubmed.ncbi.nlm.nih.gov/31845510","citation_count":4,"is_preprint":false},{"pmid":"19664060","id":"PMC_19664060","title":"Inactivating pentapeptide insertions in the fission yeast replication factor C subunit Rfc2 cluster near the ATP-binding site and arginine finger motif.","date":"2009","source":"The FEBS journal","url":"https://pubmed.ncbi.nlm.nih.gov/19664060","citation_count":3,"is_preprint":false},{"pmid":"40037867","id":"PMC_40037867","title":"Knockdown of RFC2 Prevents the Proliferation, Migration and Invasion of Cervical Cancer Cells.","date":"2025","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/40037867","citation_count":2,"is_preprint":false},{"pmid":"22720015","id":"PMC_22720015","title":"Down-regulation of replication factor C-40 (RFC40) causes chromosomal missegregation in neonatal and hypertrophic adult rat cardiac myocytes.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22720015","citation_count":2,"is_preprint":false},{"pmid":"39368701","id":"PMC_39368701","title":"RFC2 may contribute to the pathogenicity of Williams syndrome revealed in a zebrafish model.","date":"2024","source":"Journal of genetics and genomics = Yi chuan xue bao","url":"https://pubmed.ncbi.nlm.nih.gov/39368701","citation_count":1,"is_preprint":false},{"pmid":"38778443","id":"PMC_38778443","title":"RETRACTION: \"RFC2, a direct target of miR-744, modulates the cell cycle and promotes the proliferation of CRC cells\".","date":"2024","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/38778443","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":12302,"output_tokens":3250,"usd":0.042828,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10949,"output_tokens":3357,"usd":0.069335,"stage2_stop_reason":"end_turn"},"total_usd":0.112163,"stage1_batch_id":"msgbatch_01CNoDbkChHEkgrJnL7BwtGJ","stage2_batch_id":"msgbatch_016Y4x9E9Wg5VQDANj9quSQD","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2001,\n      \"finding\": \"The conserved lysine (K71) in the Walker A motif (ATP-binding domain) of yeast Rfc2 is essential for ATPase activity, DNA binding, and clamp loading. The rfc2-K71E mutation severely impairs ATPase, clamp loading, and DNA binding; the conservative rfc2-K71R mutation has milder defects suppressible at high ATP concentrations. All mutant RFC complexes retain PCNA interaction.\",\n      \"method\": \"Active-site mutagenesis (Walker A lysine → glutamate or arginine), in vitro ATPase assay, clamp-loading assay, DNA-binding assay, and PCNA-interaction assay with bacterially overproduced mutant RFC complexes\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with active-site mutagenesis and multiple orthogonal functional assays in a single rigorous study\",\n      \"pmids\": [\"11432854\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1994,\n      \"finding\": \"Yeast Rfc2 protein is a component of the RF-C complex required for DNA replication: it co-purifies with RF-C activity, and polyclonal antibodies against bacterially expressed Rfc2 specifically reduce RF-C activity in a DNA polymerase III-dependent replication reaction. Bacterially expressed Rfc2 preferentially binds primed single-stranded DNA and weakly binds ATP.\",\n      \"method\": \"Co-purification of Rfc2 with RF-C activity, antibody inhibition assay, in vitro DNA/ATP binding assay with recombinant protein, RFC2 gene disruption (lethal, dumbbell morphology)\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — multiple orthogonal methods (co-purification, antibody inhibition, in vitro binding) in a single study establishing Rfc2 as an RFC subunit\",\n      \"pmids\": [\"8202350\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"Yeast RFC2 is required for chromosomal DNA replication and an S-phase DNA damage checkpoint. The rfc2-1 thermosensitive mutation causes DNA integrity defects, sensitivity to hydroxyurea/MMS/UV, elevated mitotic recombination/chromosome loss, and checkpoint failure. Genetic epistasis shows synthetic lethality with cdc44-1/rfc1 and rfc5-1 mutations, exacerbation by cdc2-2 and pol2-11 mutations, and suppression by multicopy RFC5, indicating functional interaction with Rfc1, Rfc5, and DNA polymerases δ and ε.\",\n      \"method\": \"Thermosensitive allele (rfc2-1) genetic analysis; DNA damage sensitivity assays; mitotic recombination and chromosome loss assays; synthetic lethality and multicopy suppression screens\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal genetic methods (epistasis, suppression, synthetic lethality) in yeast with defined phenotypic readouts, replicated across multiple assays\",\n      \"pmids\": [\"9671499\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Human RFC2 is monoubiquitylated by the RAD6-RAD18 E2-E3 ubiquitin ligase complex in response to DNA-damaging agents (alkylating agents, H2O2) and this ubiquitylation is inhibited by RPA (replication protein A). A D228A mutation in RFC2 (corresponding to a yeast Rfc4 mutation that reduces RPA interaction) causes constitutive RFC2 ubiquitylation even without DNA damage, establishing that RPA binding to RFC2 suppresses its ubiquitylation. RAD6-RAD18-mediated ubiquitylation of RFC2 was reconstituted in vitro.\",\n      \"method\": \"In vitro ubiquitylation assay (RAD6-RAD18 complex + RFC2), site-directed mutagenesis (D228A), in-cell ubiquitylation assays with RAD18-dependent damage response, RPA inhibition assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution of ubiquitylation with mutagenesis and RPA inhibition, supported by in-cell validation; single lab but multiple orthogonal methods\",\n      \"pmids\": [\"18245774\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"RFC40 (RFC2) directly interacts with the regulatory subunit RIα of cAMP-dependent PKA. The interaction maps to the N-terminus of RIα and the C-terminus of RFC40. RFC37 (RFC3) competes with RIα and displaces it from the RFC40-RIα complex. RIα functions as a nuclear transport protein for RFC40, and impairment of this nuclear transport arrests cells in G1 phase.\",\n      \"method\": \"Yeast two-hybrid screening (RFC40 as bait), co-immunoprecipitation, interaction domain mapping, cell cycle analysis (flow cytometry) after nuclear transport disruption\",\n      \"journal\": \"Cancer biology & therapy / Cell cycle (Georgetown, Tex.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — yeast two-hybrid plus Co-IP for interaction, domain mapping, and functional cell cycle readout; two papers from same lab using overlapping methods\",\n      \"pmids\": [\"15655353\", \"15846072\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"CDK2/Cyclin E phosphorylates RIα at a serine residue, promoting dissociation of the RIα-RFC40 complex. In vitro phosphorylation of RIα by CDK2/CyclinE prevents its association with RFC40. CDK inhibitor olomoucine increases the RIα-RFC40 complex and decreases the RFC40-RFC37 complex. Inhibition of phosphatase PP1 by Calyculin A reduces RIα-RFC40 complex formation, consistent with phosphorylation driving dissociation.\",\n      \"method\": \"In vitro kinase assay (CDK2/CyclinE + RIα), CDK inhibitor treatment (olomoucine), phosphatase inhibitor treatment (Calyculin A), co-immunoprecipitation of complex formation\",\n      \"journal\": \"Cell cycle (Georgetown, Tex.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Weak — in vitro phosphorylation assay with pharmacological inhibitors and Co-IP; single lab, multiple methods but no direct mutagenesis of the phosphosite\",\n      \"pmids\": [\"16582606\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"In fission yeast, pentapeptide insertions in Rfc2 that abolish growth cluster near the ATP-binding sites (arginine finger motif and P-loop/sites C and D) and the central five-stranded β-sheet, as mapped onto the 3D structure of budding yeast Rfc2. Non-lethal insertions map predominantly to loop regions or outer surface of the RFC complex.\",\n      \"method\": \"Pentapeptide-scanning mutagenesis of fission yeast rfc2; growth complementation assay in rfc2Δ cells; structural mapping onto 3D structure of budding yeast Rfc2\",\n      \"journal\": \"The FEBS journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Weak — systematic mutagenesis with structural mapping; single lab, in vivo growth assay only (no in vitro biochemical activity measurements)\",\n      \"pmids\": [\"19664060\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"cAMP (via N6-monobutyryl cAMP) upregulates RFC40 mRNA and protein expression, increases RIα-RFC40 complex formation, increases the nuclear-to-cytoplasmic ratio of RFC40, and increases the proportion of S-phase cells, while decreasing RFC40-RFC37 complex formation and DNA replication rate.\",\n      \"method\": \"cAMP analog treatment of MCF7 cells; qRT-PCR; western blot; co-immunoprecipitation; nuclear/cytoplasmic fractionation; flow cytometry cell cycle analysis\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pharmacological treatment with indirect readouts; no direct mechanistic dissection of how cAMP controls RFC40\",\n      \"pmids\": [\"16413017\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"In glioblastoma cells, NELFA mRNA (noncoding mode) interacts with Rad17 and regulates the interaction between Rad17 and the RFC2-5 complex, with downstream impact on CHK1, CHK2, and BRCA1 phosphorylation in DNA damage repair signaling.\",\n      \"method\": \"Co-immunoprecipitation, RNA immunoprecipitation, knockdown experiments measuring CHK1/CHK2/BRCA1 phosphorylation\",\n      \"journal\": \"Molecular oncology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab Co-IP; RFC2 participates as part of a multi-subunit complex but its specific role is not mechanistically dissected; indirect evidence\",\n      \"pmids\": [\"31845510\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Knockdown of RFC40 (RFC2) in neonatal rat cardiac myocytes causes chromosomal missegregation/aneuploidy and decrease in cell numbers, establishing a direct role for RFC40 in maintaining chromosomal integrity during cardiac myocyte replication.\",\n      \"method\": \"siRNA knockdown of RFC40 in neonatal rat cardiac myocytes; FISH for chromosome 12 to detect missegregation/aneuploidy; cell counting\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, siRNA knockdown with FISH readout; no mechanistic pathway dissection beyond the phenotypic result\",\n      \"pmids\": [\"22720015\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"rfc2 knockout zebrafish (CRISPR-Cas9) exhibit small head/brain, jaw/dental defects, and vascular problems reminiscent of Williams syndrome. RNA-seq reveals that genes associated with neural cell survival and differentiation are specifically affected. Heterozygous rfc2 KO adult zebrafish show anxiety-like behavior with increased social cohesion.\",\n      \"method\": \"CRISPR-Cas9 knockout of rfc2 in zebrafish; phenotypic analysis; RNA-seq transcriptome analysis; behavioral assay\",\n      \"journal\": \"Journal of genetics and genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — in vivo loss-of-function with defined phenotypic readouts and transcriptomic analysis; single lab, novel organism model without biochemical mechanism\",\n      \"pmids\": [\"39368701\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RFC2 (RFC40) is the second-largest subunit of the heteropentameric Replication Factor C (RFC) complex; its Walker A ATP-binding domain (K71 in yeast) is essential for ATPase activity, DNA binding, and PCNA clamp loading onto primed DNA templates, while its arginine finger motif and central β-sheet are also structurally critical; it is subject to RAD6-RAD18-dependent monoubiquitylation after DNA damage, a modification suppressed by RPA binding to RFC2; in human cells its nuclear import depends on interaction with PKA regulatory subunit RIα (mapping to the RFC40 C-terminus), a complex whose dissociation is triggered by CDK2/CyclinE phosphorylation of RIα and is required for G1-to-S phase progression; genetic studies in yeast establish functional interactions with Rfc1, Rfc5, and DNA polymerases δ and ε, and an S-phase checkpoint role.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RFC2 (RFC40) is an essential subunit of the heteropentameric Replication Factor C (RFC) clamp-loader complex that loads the PCNA sliding clamp onto primed DNA during chromosomal replication and supports the S-phase DNA damage checkpoint [#1, #2]. Its conserved Walker A lysine (K71 in yeast) is required for ATPase activity, primed-template DNA binding, and clamp loading, while the arginine finger motif and central β-sheet are structurally critical for function [#0, #6]. RFC2 acts within a network of replication factors, showing functional interactions with Rfc1, Rfc5, and DNA polymerases δ and ε [#2]. The protein is regulated by post-translational and protein-interaction controls: it is monoubiquitylated by the RAD6-RAD18 E2-E3 ligase complex following DNA damage, a modification suppressed by RPA binding to RFC2 [#3], and in human cells its nuclear import depends on interaction with the PKA regulatory subunit RIα, a complex displaced by RFC3 (RFC37) and dissolved upon CDK2/Cyclin E phosphorylation of RIα to permit G1-to-S progression [#4, #5]. Loss of RFC2 function causes chromosomal missegregation and aneuploidy, and rfc2 knockout in zebrafish produces craniofacial, brain, and vascular defects with altered neural survival/differentiation gene expression [#9, #10].\",\n  \"teleology\": [\n    {\n      \"year\": 1994,\n      \"claim\": \"Established that Rfc2 is a bona fide subunit of the RF-C clamp-loader complex required for DNA replication, defining its place in the replication machinery.\",\n      \"evidence\": \"Co-purification with RF-C activity, antibody inhibition of a polymerase III-dependent replication reaction, in vitro DNA/ATP binding, and lethal gene disruption in yeast\",\n      \"pmids\": [\"8202350\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve the stoichiometry or catalytic mechanism within the complex\", \"ATP binding characterized only as weak with recombinant protein\"]\n    },\n    {\n      \"year\": 1998,\n      \"claim\": \"Connected RFC2 not only to bulk replication but to an S-phase DNA damage checkpoint and placed it genetically with Rfc1, Rfc5, and polymerases δ/ε.\",\n      \"evidence\": \"Thermosensitive rfc2-1 allele with DNA damage sensitivity, recombination/chromosome loss assays, synthetic lethality and multicopy suppression in yeast\",\n      \"pmids\": [\"9671499\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Genetic interactions do not establish direct physical contacts\", \"Molecular basis of the checkpoint defect not dissected\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Pinpointed the Walker A lysine K71 as essential for the ATPase, DNA-binding, and clamp-loading activities, separating catalytic function from PCNA association.\",\n      \"evidence\": \"Active-site mutagenesis (K71E/K71R) with in vitro ATPase, clamp-loading, DNA-binding, and PCNA-interaction assays on reconstituted RFC complexes\",\n      \"pmids\": [\"11432854\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not map other catalytic residues to function\", \"Mechanism of ATP-driven clamp opening not resolved\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Identified a non-replication regulatory axis: RFC40 binds PKA regulatory subunit RIα, which serves as its nuclear transport carrier, linking RFC40 nuclear import to cell cycle progression.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, domain mapping (RIα N-terminus to RFC40 C-terminus), RFC3 competition, and G1-arrest cell cycle analysis\",\n      \"pmids\": [\"15655353\", \"15846072\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether RIα transports free RFC40 or assembled RFC unclear\", \"Single lab with overlapping methods\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Showed CDK2/Cyclin E phosphorylation of RIα dissolves the RIα-RFC40 complex, providing a cell-cycle switch governing RFC40 availability.\",\n      \"evidence\": \"In vitro kinase assay, CDK inhibitor (olomoucine) and phosphatase inhibitor (Calyculin A) treatments, Co-IP of complex changes\",\n      \"pmids\": [\"16582606\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Phosphosite not confirmed by mutagenesis\", \"Single lab, pharmacological inference\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Defined a DNA-damage-responsive modification of RFC2: RAD6-RAD18 monoubiquitylation antagonized by RPA binding, coupling RFC2 to damage signaling.\",\n      \"evidence\": \"In vitro reconstituted ubiquitylation, D228A mutant causing constitutive ubiquitylation, in-cell RAD18-dependent damage response, RPA inhibition assay\",\n      \"pmids\": [\"18245774\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of the ubiquitin mark on clamp loading not established\", \"Ubiquitylation site(s) on RFC2 not mapped\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Mapped which structural elements are indispensable, confirming ATP-binding sites and the central β-sheet as functionally critical surfaces.\",\n      \"evidence\": \"Pentapeptide-scanning mutagenesis of fission yeast rfc2 with growth complementation and structural mapping onto budding yeast Rfc2\",\n      \"pmids\": [\"19664060\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"In vivo growth only; no in vitro biochemical activity measurements\", \"Effects of individual insertions on clamp loading not measured\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Demonstrated a direct cellular requirement for RFC40 in chromosomal integrity, with depletion causing missegregation and aneuploidy.\",\n      \"evidence\": \"siRNA knockdown in neonatal rat cardiac myocytes with chromosome-12 FISH and cell counting\",\n      \"pmids\": [\"22720015\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single lab phenotypic study without mechanistic dissection\", \"Off-target effects of siRNA not controlled by rescue\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Placed the RFC2-5 complex within damage-response signaling via Rad17, regulated by NELFA RNA, affecting CHK1/CHK2/BRCA1 phosphorylation.\",\n      \"evidence\": \"Co-IP, RNA immunoprecipitation, knockdown measuring checkpoint kinase phosphorylation in glioblastoma cells\",\n      \"pmids\": [\"31845510\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"RFC2-specific contribution within the complex not isolated\", \"Indirect, single-lab evidence\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Established an organismal developmental requirement, with rfc2 knockout causing craniofacial, neural, and vascular defects resembling a human syndrome.\",\n      \"evidence\": \"CRISPR-Cas9 rfc2 knockout zebrafish, phenotyping, RNA-seq, and behavioral assays\",\n      \"pmids\": [\"39368701\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No biochemical mechanism linking RFC2 loss to phenotypes\", \"Causal disease relationship in humans not established by this model\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How DNA-damage monoubiquitylation, RIα/PKA-dependent trafficking, and clamp-loading activity are integrated into a single regulatory program for RFC2 across cell cycle and development remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structure of human RFC2 within an assembled clamp-loader in the corpus\", \"Functional consequence of RFC2 monoubiquitylation undefined\", \"Connection between developmental phenotypes and replication function unestablished\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [4, 7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-69306\", \"supporting_discovery_ids\": [1, 2]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [3, 8]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [4, 5]}\n    ],\n    \"complexes\": [\"Replication Factor C (RFC) complex\"],\n    \"partners\": [\"RFC1\", \"RFC5\", \"RFC3\", \"PCNA\", \"RAD18\", \"RAD6\", \"RPA\", \"PRKAR1A\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}