{"gene":"RBPMS2","run_date":"2026-06-10T06:43:36","timeline":{"discoveries":[{"year":2012,"finding":"RBPMS2 is strongly expressed in early visceral smooth muscle cell (SMC) precursors and is rapidly downregulated upon SMC differentiation. Misexpression of RBPMS2 in differentiated visceral SMCs induces their dedifferentiation and reduces contractility by upregulating NOGGIN expression, which in turn reduces bone morphogenetic protein (BMP) activity.","method":"Avian replication-competent retroviral misexpression in vivo and in primary chicken SMC cultures; measurement of BMP activity and differentiation markers","journal":"Gastroenterology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional misexpression in vivo and in primary cells with defined pathway readout (BMP/Noggin), single lab, multiple orthogonal assays","pmids":["22683258"],"is_preprint":false},{"year":2014,"finding":"RBPMS2 homodimerizes through a conserved sequence motif (D-x-K-x-R-E-L-Y-L-L-F, residues 39–51) located within its single RRM domain. This homodimerization motif is required for RBPMS2 to interact with the translational elongation factor eEF2, to upregulate NOGGIN mRNA in vivo, and to drive SMC dedifferentiation. Mutation of a conserved leucine within this motif abolishes dimerization and all downstream functions.","method":"Structure/function analyses including RRM domain mutagenesis, co-immunoprecipitation, in vivo misexpression assays, and sequence conservation analysis across vertebrates and invertebrate orthologs","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — mutagenesis of active-site residue combined with Co-IP (eEF2 interaction), in vivo functional rescue, and phylogenetic conservation analysis; multiple orthogonal methods in one study","pmids":["25064856"],"is_preprint":false},{"year":2018,"finding":"Zebrafish Rbpms2 localizes to germ granules in primordial germ cells and to the Balbiani body of oocytes. Localization to germline compartments requires an intact RNA-binding domain, whereas localization to centrosomes/spindle in somatic blastula cells requires the conserved C-terminal domain. Loss of rbpms2a;2b leads to failure of definitive oogenesis and male sex determination; Rbpms2 is required for proper Buc protein organization within the Balbiani body.","method":"Zebrafish rbpms2a;2b double mutant generation, mutant domain-deletion protein constructs, immunohistochemistry, ultrastructural analysis (EM), live imaging of subcellular localization","journal":"PLoS genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic loss-of-function with domain-specific mutant rescue, IHC, EM ultrastructure, and multiple orthogonal methods across labs","pmids":["29975683"],"is_preprint":false},{"year":2022,"finding":"RBPMS2 functions as a conserved regulator of alternative splicing in cardiomyocytes. Rbpms2-deficient zebrafish embryos show reduced ejection fraction, myofibril disarray, and altered calcium handling, accompanied by differential alternative splicing of cardiac genes. A conserved network of 29 ortholog pairs (including RBFOX2, SLC8A1, and MYBPC3) require RBPMS2 for proper alternative splicing in both zebrafish and human iPSC-derived cardiomyocytes.","method":"Zebrafish rbpms2 knockout generation, RNA sequencing, human iPSC-derived RBPMS2-deficient cardiomyocytes, comparative alternative splicing analysis, calcium imaging","journal":"Circulation research","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO in two species (zebrafish and human iPSC-CMs), RNA-seq splicing analysis, calcium imaging, multiple orthogonal methods with cross-species replication","pmids":["36367103"],"is_preprint":false},{"year":2024,"finding":"Rbpms2 acts as a translational regulator of Rbpms2-bound oocyte RNAs (rboRNAs), repressing testis-fate factors and ribosome biogenesis factors in zebrafish oocytes. Genetically, Rbpms2 promotes nucleolar amplification via the mTORC1 pathway specifically through the Gator2 complex component Mios (Missing oocyte), functioning as a binary fate switch that promotes female over male fate.","method":"RNA immunoprecipitation to identify rboRNAs, genetic epistasis (rbpms2 and mios mutants), mTORC1 pathway analysis, polysome/translational assays in zebrafish","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — CLIP/RIP identification of targets, genetic epistasis with double mutants, pathway analysis, multiple orthogonal methods in one study with peer review","pmids":["38898112"],"is_preprint":false},{"year":2025,"finding":"RBPMS and RBPMS2 cooperate redundantly to safeguard cardiac splicing in mice. Double cardiomyocyte-specific knockout of Rbpms and Rbpms2 causes embryonic lethality before E13.5 with sarcomere disarray and widespread mis-splicing of cardiac contractile genes, whereas single knockouts survive to adulthood with normal sarcomere assembly. The binding location of RBPMS2 on pre-mRNA dictates whether it functions as a splicing activator or repressor (positional effect), and intrinsic features of target exons determine requirement for one or both proteins.","method":"Cardiomyocyte-specific single and double Rbpms/Rbpms2 knockout mice, RNA sequencing, minigene splicing assays, in silico RNA splicing map analysis","journal":"Circulation research","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional double-KO mice with embryonic lethal phenotype, RNA-seq splicing maps, minigene assays; multiple orthogonal methods with two independent publications (peer-reviewed + preprint)","pmids":["40859824","39574760"],"is_preprint":false},{"year":2025,"finding":"In Rbpms/Rbpms2 compound-mutant cardiomyocytes, mitotic defects are caused by an imbalance of nuclear versus cytoplasmic CaM-dependent protein kinase II gamma (Camk2g) isoforms due to disrupted splicing. Overexpression of the Rbpms2a isoform partially rescues these defects and prevents embryonic lethality, demonstrating that RBPMS2 regulates Camk2g isoform balance in cardiomyocytes.","method":"Rbpms/Rbpms2 compound knockout mice, cardiomyocyte-specific deletion, Rbpms2a isoform rescue overexpression, analysis of Camk2g isoform distribution (nuclear vs. cytoplasmic), embryonic lethality phenotyping","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic compound KO with isoform-specific rescue, defined mechanistic link to Camk2g isoform imbalance, embryonic lethality phenotype, multiple methods","pmids":["40602408"],"is_preprint":false},{"year":2023,"finding":"Maternal RBPMS2 is required for mouse embryo progression from the morula to blastocyst stage. Knockdown of RBPMS2 arrests embryos at the morula stage; this arrest is phenocopied by BMP pathway inhibitor and partially rescued by BMP activator. RBPMS2 knockdown impairs E-cadherin membrane localization, an effect also observed with BMP pathway inhibition, placing RBPMS2 upstream of BMP signaling in blastocyst formation.","method":"Morpholino and siRNA knockdown via microinjection in mouse embryos, RNA sequencing, BMP inhibitor/activator rescue experiments, immunofluorescence for E-cadherin localization","journal":"Reproductive biomedicine online","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function knockdown with pharmacological rescue via BMP pathway modulation, immunofluorescence readout; single lab, multiple orthogonal approaches","pmids":["37573751"],"is_preprint":false},{"year":2024,"finding":"In Japanese flounder, Rbpms2 binds to the 3'UTR, 5'UTR, and ORF of sex-related gene mRNAs (dmrt1, sox9, amh, foxl2, wnt4). Rbpms2 knockdown or overexpression in primary gonadal cells shows that Rbpms2 represses male-related genes (dmrt1, sox9, amh) and promotes female-related genes (foxl2, wnt4).","method":"siRNA knockdown and overexpression in primary ovary and testis cells, in silico binding site analysis, qRT-PCR measurement of sex-related gene expression","journal":"General and comparative endocrinology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, siRNA knockdown in primary cells with gene expression readout, no binding confirmation by RIP/CLIP, no in vivo genetic analysis","pmids":["39414089"],"is_preprint":false},{"year":2025,"finding":"RBPMS2 inhibits pyroptosis in gastric cancer cells by suppressing the NLRP3/caspase-1/GSDMD signaling pathway. Knockout of RBPMS2 activates pyroptosis, causes cell membrane damage, and increases expression of pyroptosis-related proteins. These effects are reversed by the NLRP3 inhibitor MCC950, confirming pathway specificity.","method":"RBPMS2 knockout in gastric cancer cells, western blot for NLRP3/caspase-1/GSDMD pathway proteins, pharmacological rescue with MCC950, cell membrane damage assays","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO with pharmacological rescue, defined pathway readout, multiple protein markers; single lab","pmids":["40595708"],"is_preprint":false}],"current_model":"RBPMS2 is an RRM-domain RNA-binding protein that homodimerizes via a conserved RRM motif to interact with eEF2 and regulate target mRNAs (including NOGGIN); it acts as a conserved alternative splicing regulator in cardiomyocytes (cooperating redundantly with RBPMS, with binding position on pre-mRNA determining activator vs. repressor activity), controls smooth muscle cell plasticity via the BMP/Noggin axis, promotes female fate in zebrafish oocytes by translationally repressing testis factors and activating nucleolar amplification through the Gator2-mTORC1 pathway, and in embryonic cardiomyocytes regulates Camk2g isoform balance to prevent mitotic defects."},"narrative":{"mechanistic_narrative":"RBPMS2 is an RRM-domain RNA-binding protein that controls cell-fate decisions across smooth muscle, cardiac, and germline lineages by regulating the translation and splicing of target mRNAs [PMID:25064856, PMID:36367103, PMID:38898112]. In visceral smooth muscle, RBPMS2 marks undifferentiated precursors and drives dedifferentiation by upregulating NOGGIN mRNA, thereby lowering BMP activity [PMID:22683258]; this activity depends on a conserved homodimerization motif within its single RRM domain (residues 39–51) that is also required for its interaction with the translational elongation factor eEF2, such that mutation of a single conserved leucine abolishes dimerization and all downstream functions [PMID:25064856]. In cardiomyocytes, RBPMS2 acts as a conserved alternative-splicing regulator: it is required for proper splicing of a network of cardiac genes (including RBFOX2, SLC8A1, and MYBPC3) in both zebrafish and human iPSC-derived cardiomyocytes [PMID:36367103], and it cooperates redundantly with its paralog RBPMS to safeguard sarcomere assembly, with the binding position on pre-mRNA dictating whether RBPMS2 acts as a splicing activator or repressor [PMID:40859824, PMID:39574760]. One critical splicing output is the balance of nuclear versus cytoplasmic Camk2g isoforms, whose disruption causes mitotic defects rescued by the Rbpms2a isoform [PMID:40602408]. In zebrafish oocytes, RBPMS2 localizes to germ granules and the Balbiani body and functions as a binary female-fate switch, translationally repressing testis-fate and ribosome-biogenesis factors while promoting nucleolar amplification through the Gator2 component Mios and the mTORC1 pathway [PMID:29975683, PMID:38898112].","teleology":[{"year":2012,"claim":"Established that RBPMS2 is not merely a precursor marker but an active determinant of smooth muscle cell plasticity, working through the NOGGIN/BMP axis.","evidence":"Retroviral misexpression in vivo and in primary chicken SMC cultures with BMP-activity and differentiation-marker readouts","pmids":["22683258"],"confidence":"Medium","gaps":["Mechanism by which RBPMS2 upregulates NOGGIN (translational vs. transcriptional) not resolved","Single-lab functional study without direct RNA-target identification"]},{"year":2014,"claim":"Defined the structural basis of RBPMS2 activity, showing a conserved RRM-internal homodimerization motif couples self-association to eEF2 binding and to NOGGIN regulation.","evidence":"RRM mutagenesis, co-immunoprecipitation of eEF2, in vivo misexpression rescue, and cross-species conservation analysis","pmids":["25064856"],"confidence":"High","gaps":["No high-resolution structure of the dimer or the eEF2 complex","How eEF2 binding translates into NOGGIN mRNA regulation is not mechanistically defined"]},{"year":2018,"claim":"Resolved domain-specific subcellular targeting and germline function, separating RNA-binding-dependent germ-granule/Balbiani-body localization from C-terminal-dependent centrosome/spindle localization.","evidence":"Zebrafish rbpms2a;2b double mutants, domain-deletion rescue constructs, immunohistochemistry, EM, and live imaging","pmids":["29975683"],"confidence":"High","gaps":["RNA cargo localized to germ granules not identified in this study","Role of centrosome/spindle localization in somatic cells unexplained"]},{"year":2022,"claim":"Identified RBPMS2 as a conserved cardiomyocyte alternative-splicing regulator, linking its loss to contractile dysfunction across zebrafish and human cells.","evidence":"Zebrafish knockout, RNA-seq splicing analysis, human iPSC-derived RBPMS2-deficient cardiomyocytes, and calcium imaging","pmids":["36367103"],"confidence":"High","gaps":["Direct binding sites on cardiac targets not mapped","Relationship between splicing role and earlier eEF2/translational role unclear"]},{"year":2023,"claim":"Extended RBPMS2 function to early mammalian development, placing it upstream of BMP signaling in the morula-to-blastocyst transition.","evidence":"Morpholino/siRNA knockdown in mouse embryos, RNA-seq, BMP inhibitor/activator rescue, and E-cadherin immunofluorescence","pmids":["37573751"],"confidence":"Medium","gaps":["Direct RNA targets mediating the BMP and E-cadherin effects not identified","Knockdown rather than genetic deletion"]},{"year":2024,"claim":"Defined the molecular logic of RBPMS2's female-fate decision in oocytes, connecting translational repression of testis factors to mTORC1-driven nucleolar amplification via Gator2/Mios.","evidence":"RNA immunoprecipitation to identify bound rboRNAs, genetic epistasis of rbpms2 and mios mutants, and polysome/translational assays in zebrafish","pmids":["38898112"],"confidence":"High","gaps":["How RBPMS2 mechanistically activates mTORC1 through Mios not resolved","Sequence determinants distinguishing repressed targets not defined"]},{"year":2024,"claim":"Tested whether the sex-fate role generalizes to teleost gonadal cells, showing Rbpms2 binds sex-gene transcripts and biases expression toward female fate.","evidence":"siRNA knockdown and overexpression in primary flounder gonadal cells, in silico binding-site analysis, and qRT-PCR","pmids":["39414089"],"confidence":"Low","gaps":["No binding confirmation by RIP/CLIP; binding inferred in silico only","No in vivo genetic validation","Cannot distinguish direct regulation from indirect effects"]},{"year":2025,"claim":"Demonstrated functional redundancy with the paralog RBPMS in cardiac splicing and established a positional code in which pre-mRNA binding location dictates activator versus repressor activity.","evidence":"Cardiomyocyte-specific single and double Rbpms/Rbpms2 knockout mice, RNA-seq splicing maps, and minigene assays","pmids":["40859824","39574760"],"confidence":"High","gaps":["Structural basis of positional activator/repressor switching not defined","Features of target exons requiring one versus both proteins only partly characterized"]},{"year":2025,"claim":"Pinpointed a specific splicing output—Camk2g nuclear/cytoplasmic isoform balance—as the cause of mitotic defects in cardiomyocytes lacking RBPMS proteins.","evidence":"Compound knockout mice with cardiomyocyte deletion and Rbpms2a isoform rescue, plus analysis of Camk2g isoform distribution and embryonic lethality","pmids":["40602408"],"confidence":"High","gaps":["How Camk2g isoform imbalance mechanistically triggers mitotic failure not fully resolved","Whether other splicing targets contribute to the phenotype unclear"]},{"year":2025,"claim":"Implicated RBPMS2 in suppressing cancer-cell death, linking it to inhibition of NLRP3/caspase-1/GSDMD pyroptosis in gastric cancer.","evidence":"RBPMS2 knockout in gastric cancer cells, western blotting of pyroptosis pathway proteins, MCC950 pharmacological rescue, and membrane-damage assays","pmids":["40595708"],"confidence":"Medium","gaps":["Direct RNA target connecting RBPMS2 to NLRP3 not identified","Whether effect is splicing- or translation-mediated unknown"]},{"year":null,"claim":"How RBPMS2's biochemical activities—RRM-dependent RNA binding, homodimerization, and eEF2 interaction—are mechanistically partitioned between translational regulation and alternative splicing across its diverse tissue contexts remains unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No unified model reconciling eEF2-coupled translational control with splicing-factor activity","Tissue-specific cofactors directing each activity not identified","No experimental structure of RBPMS2 bound to RNA or partner proteins"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003723","term_label":"RNA binding","supporting_discovery_ids":[1,2,4,8]},{"term_id":"GO:0045182","term_label":"translation regulator activity","supporting_discovery_ids":[4]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[3,5]}],"localization":[{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[2]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[2]}],"pathway":[{"term_id":"R-HSA-8953854","term_label":"Metabolism of RNA","supporting_discovery_ids":[3,5]},{"term_id":"R-HSA-1474165","term_label":"Reproduction","supporting_discovery_ids":[2,4]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,7]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[0,3]}],"complexes":[],"partners":["EEF2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q6ZRY4","full_name":"RNA-binding protein with multiple splicing 2","aliases":["RNA binding protein, mRNA processing factor 2"],"length_aa":209,"mass_kda":22.5,"function":"RNA-binding protein involved in the regulation of smooth muscle cell differentiation and proliferation in the gastrointestinal system (PubMed:25064856). Binds NOG mRNA, the major inhibitor of the bone morphogenetic protein (BMP) pathway. Mediates an increase of NOG mRNA levels, thereby contributing to the negative regulation of BMP signaling pathway and promoting reversible dedifferentiation and proliferation of smooth muscle cells (By similarity). Acts as a pre-mRNA alternative splicing regulator (By similarity). Mediates ACTN1 and FLNB alternative splicing (By similarity). Likely binds to mRNA tandem CAC trinucleotide or CA dinucleotide motifs (PubMed:24860013)","subcellular_location":"Cytoplasm; Nucleus; Cytoplasm, Stress granule","url":"https://www.uniprot.org/uniprotkb/Q6ZRY4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RBPMS2","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/RBPMS2","total_profiled":1310},"omim":[{"mim_id":"619034","title":"RNA-BINDING PROTEIN, mRNA-PROCESSING FACTOR, 2; RBPMS2","url":"https://www.omim.org/entry/619034"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"blood vessel","ntpm":138.0},{"tissue":"heart muscle","ntpm":283.6},{"tissue":"intestine","ntpm":138.0}],"url":"https://www.proteinatlas.org/search/RBPMS2"},"hgnc":{"alias_symbol":[],"prev_symbol":[]},"alphafold":{"accession":"Q6ZRY4","domains":[{"cath_id":"3.30.70.330","chopping":"33-106","consensus_level":"high","plddt":96.2001,"start":33,"end":106}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q6ZRY4","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q6ZRY4-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q6ZRY4-F1-predicted_aligned_error_v6.png","plddt_mean":68.38},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RBPMS2","jax_strain_url":"https://www.jax.org/strain/search?query=RBPMS2"},"sequence":{"accession":"Q6ZRY4","fasta_url":"https://rest.uniprot.org/uniprotkb/Q6ZRY4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q6ZRY4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q6ZRY4"}},"corpus_meta":[{"pmid":"22683258","id":"PMC_22683258","title":"The RNA-binding protein RBPMS2 regulates development of gastrointestinal smooth muscle.","date":"2012","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/22683258","citation_count":37,"is_preprint":false},{"pmid":"29975683","id":"PMC_29975683","title":"rbpms2 functions in Balbiani body architecture and ovary fate.","date":"2018","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/29975683","citation_count":36,"is_preprint":false},{"pmid":"25064856","id":"PMC_25064856","title":"Homodimerization of RBPMS2 through a new RRM-interaction motif is necessary to control smooth muscle plasticity.","date":"2014","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/25064856","citation_count":29,"is_preprint":false},{"pmid":"23295309","id":"PMC_23295309","title":"High expression of the RNA-binding protein RBPMS2 in gastrointestinal stromal tumors.","date":"2013","source":"Experimental and molecular pathology","url":"https://pubmed.ncbi.nlm.nih.gov/23295309","citation_count":26,"is_preprint":false},{"pmid":"36367103","id":"PMC_36367103","title":"RBPMS2 Is a Myocardial-Enriched Splicing Regulator Required for Cardiac Function.","date":"2022","source":"Circulation research","url":"https://pubmed.ncbi.nlm.nih.gov/36367103","citation_count":25,"is_preprint":false},{"pmid":"38898112","id":"PMC_38898112","title":"Rbpms2 promotes female fate upstream of the nutrient sensing Gator2 complex component Mios.","date":"2024","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/38898112","citation_count":12,"is_preprint":false},{"pmid":"34981466","id":"PMC_34981466","title":"RBPMS2, as a novel biomarker for predicting lymph node metastasis, guides therapeutic regimens in gastric cancer.","date":"2022","source":"Human cell","url":"https://pubmed.ncbi.nlm.nih.gov/34981466","citation_count":7,"is_preprint":false},{"pmid":"40602408","id":"PMC_40602408","title":"Rbpms2 prevents major cardiac defects in cardiomyocyte-specific Rbpms-deficient mice.","date":"2025","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/40602408","citation_count":2,"is_preprint":false},{"pmid":"40595708","id":"PMC_40595708","title":"RBPMS2 can inhibit the NLRP3 / caspase-1 / GSDMD signaling pathway to resist pyroptosis in gastric cancer cells.","date":"2025","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/40595708","citation_count":2,"is_preprint":false},{"pmid":"35675541","id":"PMC_35675541","title":"WNT1 expression influences the development of dysplasia of the hip via regulating RBPMS2/NOG-BMP2/4-GDF5- WISP2 pathway.","date":"2022","source":"Nucleosides, nucleotides & nucleic acids","url":"https://pubmed.ncbi.nlm.nih.gov/35675541","citation_count":2,"is_preprint":false},{"pmid":"37573751","id":"PMC_37573751","title":"Maternal RNA binding protein with multiple splicing 2 (RBPMS2) is involved in mouse blastocyst formation through the bone morphogenetic protein pathway.","date":"2023","source":"Reproductive biomedicine online","url":"https://pubmed.ncbi.nlm.nih.gov/37573751","citation_count":1,"is_preprint":false},{"pmid":"40859824","id":"PMC_40859824","title":"RBPMS and RBPMS2 Cooperate to Safeguard Cardiac Splicing.","date":"2025","source":"Circulation research","url":"https://pubmed.ncbi.nlm.nih.gov/40859824","citation_count":0,"is_preprint":false},{"pmid":"39414089","id":"PMC_39414089","title":"Identification, expression, and function analysis of Rbpms2 splicing variants in Japanese flounder gonad.","date":"2024","source":"General and comparative endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/39414089","citation_count":0,"is_preprint":false},{"pmid":"39574760","id":"PMC_39574760","title":"RBPMS and RBPMS2 Cooperate to Safeguard Cardiac Splicing.","date":"2024","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/39574760","citation_count":0,"is_preprint":false},{"pmid":"38328218","id":"PMC_38328218","title":"Rbpms2 promotes female fate upstream of the nutrient sensing Gator2 complex component, Mios.","date":"2024","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/38328218","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.03.19.644129","title":"Differential retinal ganglion cell resilience to optic nerve injury across vertebrate species","date":"2025-03-19","source":"bioRxiv","url":"https://doi.org/10.1101/2025.03.19.644129","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11143,"output_tokens":3066,"usd":0.03971,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10480,"output_tokens":3752,"usd":0.0731,"stage2_stop_reason":"end_turn"},"total_usd":0.11281,"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\": 2012,\n      \"finding\": \"RBPMS2 is strongly expressed in early visceral smooth muscle cell (SMC) precursors and is rapidly downregulated upon SMC differentiation. Misexpression of RBPMS2 in differentiated visceral SMCs induces their dedifferentiation and reduces contractility by upregulating NOGGIN expression, which in turn reduces bone morphogenetic protein (BMP) activity.\",\n      \"method\": \"Avian replication-competent retroviral misexpression in vivo and in primary chicken SMC cultures; measurement of BMP activity and differentiation markers\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional misexpression in vivo and in primary cells with defined pathway readout (BMP/Noggin), single lab, multiple orthogonal assays\",\n      \"pmids\": [\"22683258\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"RBPMS2 homodimerizes through a conserved sequence motif (D-x-K-x-R-E-L-Y-L-L-F, residues 39–51) located within its single RRM domain. This homodimerization motif is required for RBPMS2 to interact with the translational elongation factor eEF2, to upregulate NOGGIN mRNA in vivo, and to drive SMC dedifferentiation. Mutation of a conserved leucine within this motif abolishes dimerization and all downstream functions.\",\n      \"method\": \"Structure/function analyses including RRM domain mutagenesis, co-immunoprecipitation, in vivo misexpression assays, and sequence conservation analysis across vertebrates and invertebrate orthologs\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — mutagenesis of active-site residue combined with Co-IP (eEF2 interaction), in vivo functional rescue, and phylogenetic conservation analysis; multiple orthogonal methods in one study\",\n      \"pmids\": [\"25064856\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Zebrafish Rbpms2 localizes to germ granules in primordial germ cells and to the Balbiani body of oocytes. Localization to germline compartments requires an intact RNA-binding domain, whereas localization to centrosomes/spindle in somatic blastula cells requires the conserved C-terminal domain. Loss of rbpms2a;2b leads to failure of definitive oogenesis and male sex determination; Rbpms2 is required for proper Buc protein organization within the Balbiani body.\",\n      \"method\": \"Zebrafish rbpms2a;2b double mutant generation, mutant domain-deletion protein constructs, immunohistochemistry, ultrastructural analysis (EM), live imaging of subcellular localization\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic loss-of-function with domain-specific mutant rescue, IHC, EM ultrastructure, and multiple orthogonal methods across labs\",\n      \"pmids\": [\"29975683\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RBPMS2 functions as a conserved regulator of alternative splicing in cardiomyocytes. Rbpms2-deficient zebrafish embryos show reduced ejection fraction, myofibril disarray, and altered calcium handling, accompanied by differential alternative splicing of cardiac genes. A conserved network of 29 ortholog pairs (including RBFOX2, SLC8A1, and MYBPC3) require RBPMS2 for proper alternative splicing in both zebrafish and human iPSC-derived cardiomyocytes.\",\n      \"method\": \"Zebrafish rbpms2 knockout generation, RNA sequencing, human iPSC-derived RBPMS2-deficient cardiomyocytes, comparative alternative splicing analysis, calcium imaging\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO in two species (zebrafish and human iPSC-CMs), RNA-seq splicing analysis, calcium imaging, multiple orthogonal methods with cross-species replication\",\n      \"pmids\": [\"36367103\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Rbpms2 acts as a translational regulator of Rbpms2-bound oocyte RNAs (rboRNAs), repressing testis-fate factors and ribosome biogenesis factors in zebrafish oocytes. Genetically, Rbpms2 promotes nucleolar amplification via the mTORC1 pathway specifically through the Gator2 complex component Mios (Missing oocyte), functioning as a binary fate switch that promotes female over male fate.\",\n      \"method\": \"RNA immunoprecipitation to identify rboRNAs, genetic epistasis (rbpms2 and mios mutants), mTORC1 pathway analysis, polysome/translational assays in zebrafish\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — CLIP/RIP identification of targets, genetic epistasis with double mutants, pathway analysis, multiple orthogonal methods in one study with peer review\",\n      \"pmids\": [\"38898112\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RBPMS and RBPMS2 cooperate redundantly to safeguard cardiac splicing in mice. Double cardiomyocyte-specific knockout of Rbpms and Rbpms2 causes embryonic lethality before E13.5 with sarcomere disarray and widespread mis-splicing of cardiac contractile genes, whereas single knockouts survive to adulthood with normal sarcomere assembly. The binding location of RBPMS2 on pre-mRNA dictates whether it functions as a splicing activator or repressor (positional effect), and intrinsic features of target exons determine requirement for one or both proteins.\",\n      \"method\": \"Cardiomyocyte-specific single and double Rbpms/Rbpms2 knockout mice, RNA sequencing, minigene splicing assays, in silico RNA splicing map analysis\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional double-KO mice with embryonic lethal phenotype, RNA-seq splicing maps, minigene assays; multiple orthogonal methods with two independent publications (peer-reviewed + preprint)\",\n      \"pmids\": [\"40859824\", \"39574760\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In Rbpms/Rbpms2 compound-mutant cardiomyocytes, mitotic defects are caused by an imbalance of nuclear versus cytoplasmic CaM-dependent protein kinase II gamma (Camk2g) isoforms due to disrupted splicing. Overexpression of the Rbpms2a isoform partially rescues these defects and prevents embryonic lethality, demonstrating that RBPMS2 regulates Camk2g isoform balance in cardiomyocytes.\",\n      \"method\": \"Rbpms/Rbpms2 compound knockout mice, cardiomyocyte-specific deletion, Rbpms2a isoform rescue overexpression, analysis of Camk2g isoform distribution (nuclear vs. cytoplasmic), embryonic lethality phenotyping\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic compound KO with isoform-specific rescue, defined mechanistic link to Camk2g isoform imbalance, embryonic lethality phenotype, multiple methods\",\n      \"pmids\": [\"40602408\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Maternal RBPMS2 is required for mouse embryo progression from the morula to blastocyst stage. Knockdown of RBPMS2 arrests embryos at the morula stage; this arrest is phenocopied by BMP pathway inhibitor and partially rescued by BMP activator. RBPMS2 knockdown impairs E-cadherin membrane localization, an effect also observed with BMP pathway inhibition, placing RBPMS2 upstream of BMP signaling in blastocyst formation.\",\n      \"method\": \"Morpholino and siRNA knockdown via microinjection in mouse embryos, RNA sequencing, BMP inhibitor/activator rescue experiments, immunofluorescence for E-cadherin localization\",\n      \"journal\": \"Reproductive biomedicine online\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function knockdown with pharmacological rescue via BMP pathway modulation, immunofluorescence readout; single lab, multiple orthogonal approaches\",\n      \"pmids\": [\"37573751\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In Japanese flounder, Rbpms2 binds to the 3'UTR, 5'UTR, and ORF of sex-related gene mRNAs (dmrt1, sox9, amh, foxl2, wnt4). Rbpms2 knockdown or overexpression in primary gonadal cells shows that Rbpms2 represses male-related genes (dmrt1, sox9, amh) and promotes female-related genes (foxl2, wnt4).\",\n      \"method\": \"siRNA knockdown and overexpression in primary ovary and testis cells, in silico binding site analysis, qRT-PCR measurement of sex-related gene expression\",\n      \"journal\": \"General and comparative endocrinology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, siRNA knockdown in primary cells with gene expression readout, no binding confirmation by RIP/CLIP, no in vivo genetic analysis\",\n      \"pmids\": [\"39414089\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RBPMS2 inhibits pyroptosis in gastric cancer cells by suppressing the NLRP3/caspase-1/GSDMD signaling pathway. Knockout of RBPMS2 activates pyroptosis, causes cell membrane damage, and increases expression of pyroptosis-related proteins. These effects are reversed by the NLRP3 inhibitor MCC950, confirming pathway specificity.\",\n      \"method\": \"RBPMS2 knockout in gastric cancer cells, western blot for NLRP3/caspase-1/GSDMD pathway proteins, pharmacological rescue with MCC950, cell membrane damage assays\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with pharmacological rescue, defined pathway readout, multiple protein markers; single lab\",\n      \"pmids\": [\"40595708\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RBPMS2 is an RRM-domain RNA-binding protein that homodimerizes via a conserved RRM motif to interact with eEF2 and regulate target mRNAs (including NOGGIN); it acts as a conserved alternative splicing regulator in cardiomyocytes (cooperating redundantly with RBPMS, with binding position on pre-mRNA determining activator vs. repressor activity), controls smooth muscle cell plasticity via the BMP/Noggin axis, promotes female fate in zebrafish oocytes by translationally repressing testis factors and activating nucleolar amplification through the Gator2-mTORC1 pathway, and in embryonic cardiomyocytes regulates Camk2g isoform balance to prevent mitotic defects.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RBPMS2 is an RRM-domain RNA-binding protein that controls cell-fate decisions across smooth muscle, cardiac, and germline lineages by regulating the translation and splicing of target mRNAs [#1, #3, #4]. In visceral smooth muscle, RBPMS2 marks undifferentiated precursors and drives dedifferentiation by upregulating NOGGIN mRNA, thereby lowering BMP activity [#0]; this activity depends on a conserved homodimerization motif within its single RRM domain (residues 39\\u201351) that is also required for its interaction with the translational elongation factor eEF2, such that mutation of a single conserved leucine abolishes dimerization and all downstream functions [#1]. In cardiomyocytes, RBPMS2 acts as a conserved alternative-splicing regulator: it is required for proper splicing of a network of cardiac genes (including RBFOX2, SLC8A1, and MYBPC3) in both zebrafish and human iPSC-derived cardiomyocytes [#3], and it cooperates redundantly with its paralog RBPMS to safeguard sarcomere assembly, with the binding position on pre-mRNA dictating whether RBPMS2 acts as a splicing activator or repressor [#5]. One critical splicing output is the balance of nuclear versus cytoplasmic Camk2g isoforms, whose disruption causes mitotic defects rescued by the Rbpms2a isoform [#6]. In zebrafish oocytes, RBPMS2 localizes to germ granules and the Balbiani body and functions as a binary female-fate switch, translationally repressing testis-fate and ribosome-biogenesis factors while promoting nucleolar amplification through the Gator2 component Mios and the mTORC1 pathway [#2, #4].\",\n  \"teleology\": [\n    {\n      \"year\": 2012,\n      \"claim\": \"Established that RBPMS2 is not merely a precursor marker but an active determinant of smooth muscle cell plasticity, working through the NOGGIN/BMP axis.\",\n      \"evidence\": \"Retroviral misexpression in vivo and in primary chicken SMC cultures with BMP-activity and differentiation-marker readouts\",\n      \"pmids\": [\"22683258\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Mechanism by which RBPMS2 upregulates NOGGIN (translational vs. transcriptional) not resolved\", \"Single-lab functional study without direct RNA-target identification\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Defined the structural basis of RBPMS2 activity, showing a conserved RRM-internal homodimerization motif couples self-association to eEF2 binding and to NOGGIN regulation.\",\n      \"evidence\": \"RRM mutagenesis, co-immunoprecipitation of eEF2, in vivo misexpression rescue, and cross-species conservation analysis\",\n      \"pmids\": [\"25064856\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No high-resolution structure of the dimer or the eEF2 complex\", \"How eEF2 binding translates into NOGGIN mRNA regulation is not mechanistically defined\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Resolved domain-specific subcellular targeting and germline function, separating RNA-binding-dependent germ-granule/Balbiani-body localization from C-terminal-dependent centrosome/spindle localization.\",\n      \"evidence\": \"Zebrafish rbpms2a;2b double mutants, domain-deletion rescue constructs, immunohistochemistry, EM, and live imaging\",\n      \"pmids\": [\"29975683\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"RNA cargo localized to germ granules not identified in this study\", \"Role of centrosome/spindle localization in somatic cells unexplained\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identified RBPMS2 as a conserved cardiomyocyte alternative-splicing regulator, linking its loss to contractile dysfunction across zebrafish and human cells.\",\n      \"evidence\": \"Zebrafish knockout, RNA-seq splicing analysis, human iPSC-derived RBPMS2-deficient cardiomyocytes, and calcium imaging\",\n      \"pmids\": [\"36367103\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct binding sites on cardiac targets not mapped\", \"Relationship between splicing role and earlier eEF2/translational role unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended RBPMS2 function to early mammalian development, placing it upstream of BMP signaling in the morula-to-blastocyst transition.\",\n      \"evidence\": \"Morpholino/siRNA knockdown in mouse embryos, RNA-seq, BMP inhibitor/activator rescue, and E-cadherin immunofluorescence\",\n      \"pmids\": [\"37573751\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct RNA targets mediating the BMP and E-cadherin effects not identified\", \"Knockdown rather than genetic deletion\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined the molecular logic of RBPMS2's female-fate decision in oocytes, connecting translational repression of testis factors to mTORC1-driven nucleolar amplification via Gator2/Mios.\",\n      \"evidence\": \"RNA immunoprecipitation to identify bound rboRNAs, genetic epistasis of rbpms2 and mios mutants, and polysome/translational assays in zebrafish\",\n      \"pmids\": [\"38898112\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"How RBPMS2 mechanistically activates mTORC1 through Mios not resolved\", \"Sequence determinants distinguishing repressed targets not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Tested whether the sex-fate role generalizes to teleost gonadal cells, showing Rbpms2 binds sex-gene transcripts and biases expression toward female fate.\",\n      \"evidence\": \"siRNA knockdown and overexpression in primary flounder gonadal cells, in silico binding-site analysis, and qRT-PCR\",\n      \"pmids\": [\"39414089\"],\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No binding confirmation by RIP/CLIP; binding inferred in silico only\", \"No in vivo genetic validation\", \"Cannot distinguish direct regulation from indirect effects\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Demonstrated functional redundancy with the paralog RBPMS in cardiac splicing and established a positional code in which pre-mRNA binding location dictates activator versus repressor activity.\",\n      \"evidence\": \"Cardiomyocyte-specific single and double Rbpms/Rbpms2 knockout mice, RNA-seq splicing maps, and minigene assays\",\n      \"pmids\": [\"40859824\", \"39574760\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Structural basis of positional activator/repressor switching not defined\", \"Features of target exons requiring one versus both proteins only partly characterized\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Pinpointed a specific splicing output—Camk2g nuclear/cytoplasmic isoform balance—as the cause of mitotic defects in cardiomyocytes lacking RBPMS proteins.\",\n      \"evidence\": \"Compound knockout mice with cardiomyocyte deletion and Rbpms2a isoform rescue, plus analysis of Camk2g isoform distribution and embryonic lethality\",\n      \"pmids\": [\"40602408\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"How Camk2g isoform imbalance mechanistically triggers mitotic failure not fully resolved\", \"Whether other splicing targets contribute to the phenotype unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Implicated RBPMS2 in suppressing cancer-cell death, linking it to inhibition of NLRP3/caspase-1/GSDMD pyroptosis in gastric cancer.\",\n      \"evidence\": \"RBPMS2 knockout in gastric cancer cells, western blotting of pyroptosis pathway proteins, MCC950 pharmacological rescue, and membrane-damage assays\",\n      \"pmids\": [\"40595708\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct RNA target connecting RBPMS2 to NLRP3 not identified\", \"Whether effect is splicing- or translation-mediated unknown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How RBPMS2's biochemical activities—RRM-dependent RNA binding, homodimerization, and eEF2 interaction—are mechanistically partitioned between translational regulation and alternative splicing across its diverse tissue contexts remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No unified model reconciling eEF2-coupled translational control with splicing-factor activity\", \"Tissue-specific cofactors directing each activity not identified\", \"No experimental structure of RBPMS2 bound to RNA or partner proteins\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003723\", \"supporting_discovery_ids\": [1, 2, 4, 8]},\n      {\"term_id\": \"GO:0045182\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [3, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-8953854\", \"supporting_discovery_ids\": [3, 5]},\n      {\"term_id\": \"R-HSA-1474165\", \"supporting_discovery_ids\": [2, 4]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 7]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [0, 3]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"eEF2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}