{"gene":"RHOT2","run_date":"2026-06-10T06:43:36","timeline":{"discoveries":[{"year":2018,"finding":"Miro1 and Miro2 were identified as mitochondrial receptors for myosin XIX (Myo19); Miro1 binds directly to a C-terminal fragment of the Myo19 tail region, and Miro1/2 recruit the Myo19 tail in vivo. This recruitment is regulated by the nucleotide state of the N-terminal Rho-like GTPase domain of Miro1/2. Myo19 protein stability in cells depends on its association with Miro1/2.","method":"Proximity labelling, direct interaction/pulldown studies, in vivo recruitment assays, downregulation experiments","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal interaction studies, in vivo recruitment, mechanistic dissection of GTPase domain regulation, replicated across Miro1 and Miro2","pmids":["30111583"],"is_preprint":false},{"year":2020,"finding":"Miro1 and Miro2 localize to peroxisomes (in addition to mitochondria), where they negatively regulate Drp1-dependent peroxisomal fission to maintain peroxisomal size and morphology. Peroxisomal localization of Miro is regulated by its first GTPase domain and mediated by an interaction of its transmembrane domain with the peroxisomal-membrane protein chaperone Pex19.","method":"Localization studies, genetic knockdown/knockout with peroxisomal morphology readout, interaction assays (Pex19 binding)","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 2 / Moderate — direct localization experiments with functional readout, identified binding partner (Pex19), domain mapping via GTPase domain and transmembrane domain, multiple orthogonal methods in one study","pmids":["31894645"],"is_preprint":false},{"year":2019,"finding":"Miro2 regulates inter-mitochondrial communication in cardiomyocytes by promoting mitochondrial nanotunneling and kissing along microtubules. Parkin-mediated ubiquitination leads to degradation of Miro2, and proteasome inhibition blocked phenylephrine-induced decrease of Miro2, establishing Parkin as the E3 ubiquitin ligase that targets Miro2 for degradation during hypertrophic stress.","method":"Adenovirus-mediated overexpression, transgenic mice, mitochondria-targeted photoactivatable GFP for communication assay, proteasome inhibition, Parkin overexpression","journal":"Circulation research","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (live imaging, genetic models, pharmacological inhibition), functional phenotype with defined molecular mechanism (Parkin ubiquitination)","pmids":["31455181"],"is_preprint":false},{"year":2019,"finding":"Miro2 undergoes CCCP-induced demultimerization from a tetramer to a monomer and changes mitochondrial localization (realignment). This realignment is regulated by PINK1-mediated phosphorylation at Ser325/Ser430 and by Ca2+ binding to the EF2 domain, both of which are required for subsequent Parkin translocation to damaged mitochondria during mitophagy.","method":"CCCP treatment, phosphorylation site mutagenesis, Ca2+ binding domain analysis, mitophagy/Parkin translocation assays, Miro2 knockout mouse phenotyping","journal":"Science bulletin","confidence":"High","confidence_rationale":"Tier 2 / Moderate — site-specific mutagenesis, defined post-translational modification (PINK1 phosphorylation), Ca2+ binding domain functional validation, in vivo knockout phenotype","pmids":["36659543"],"is_preprint":false},{"year":2022,"finding":"MIRO2 interacts with General Control Nonderepressible 1 (GCN1) in prostate cancer cells, and this interaction is necessary for efficient GCN1-mediated GCN2 kinase signaling and induction of ATF4. MIRO2 mutation 159L increases GCN1 binding. MIRO2's effect on prostate cancer cell growth is mediated through ATF4.","method":"Co-immunoprecipitation, network analysis of binding partners, functional mutagenesis (MIRO2 159L), siRNA knockdown, xenograft tumor models","journal":"Molecular cancer research : MCR","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP interaction, mutagenesis identifying enhanced binding variant, downstream signaling readout (GCN2/ATF4), single lab","pmids":["34992146"],"is_preprint":false},{"year":2020,"finding":"miR-351-5p targets Miro2 in hippocampal neural progenitor cells; Miro2 knockdown induces excessive mitochondrial fragmentation, mitochondrial dysfunction (decreased membrane potential, increased ROS), and cell death with subsequent PINK1/Parkin-mediated mitophagy induction. Suppression of mitochondrial fission by Mdivi-1 completely inhibited miR-351-5p-induced cell death, placing Miro2 upstream of Drp1-dependent fission.","method":"siRNA knockdown, adenovirus rescue, Mdivi-1 pharmacological inhibition, mitochondrial morphology and function assays","journal":"Molecular therapy. Nucleic acids","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with rescue experiment, pharmacological epistasis placing Miro2 upstream of Drp1 fission, multiple functional readouts","pmids":["33575111"],"is_preprint":false},{"year":2024,"finding":"MIRO2 supports tumor cell invasion and metastasis through cooperation with unconventional myosin 9B (MYO9B); depletion of MIRO2 phenocopies MYO9B depletion (reduced invasion and increased active RhoA), and dual ablation of MIRO2 and RhoA fully rescues tumor cell invasion. MIRO2 is required for MYO9B-driven invasion.","method":"siRNA knockdown, double-gene ablation (MIRO2 + RhoA), in vitro invasion assays, mouse metastasis models, RhoA activity assays","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis via double ablation rescue experiment, phenotypic rescue and in vivo model, single lab","pmids":["39723893"],"is_preprint":false},{"year":2024,"finding":"Miro2 cysteines C185 and C504 are sulfhydrated by CBS-derived H2S; this sulfhydration is required for maintenance of mitochondrial dynamics and for trophoblast invasion and migration. Double mutation of Miro2 C185/C504 to serine fragmented mitochondria and inhibited invasion/migration that could not be rescued by H2S donor treatment.","method":"CBS knockdown, H2S donor (GYY4137) treatment, site-directed mutagenesis (C185S/C504S double mutant), mitochondrial morphology assays, invasion/migration assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — mutagenesis of specific sulfhydration sites with functional rescue/loss readout, single lab, single study","pmids":["39461943"],"is_preprint":false},{"year":2025,"finding":"Knockdown of Miro2, but not Miro1, reduces mitochondrial motility specifically in pancreatic alpha cells and impairs glucose-induced inhibition of glucagon secretion, without affecting insulin secretion or mitochondrial motility in non-alpha islet cells. Under low glucose, mitochondria are arrested in positions further from the nucleus, correlating with increased ATP/ADP in the sub-plasma membrane space.","method":"siRNA knockdown of Miro2 vs Miro1, mitochondrial motility imaging, glucagon/insulin secretion assays, ATP/ADP measurements","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — loss-of-function with specific functional readout and isoform selectivity established, single lab, single study","pmids":["41308986"],"is_preprint":false},{"year":2025,"finding":"MIRO2 in cancer cells is required for mitochondrial transfer to fibroblasts; depletion of MIRO2 in cancer cells suppresses mitochondrial transfer and inhibits cancer-associated fibroblast (CAF) differentiation and tumor growth.","method":"MIRO2 depletion in cancer cells, co-culture and xenograft assays, mitochondrial transfer quantification, CAF marker analysis","journal":"Nature cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with defined cellular phenotype (blocked mitochondrial transfer), in vitro and in vivo models, single lab","pmids":["40877413"],"is_preprint":false},{"year":2022,"finding":"NMR backbone resonance assignments of the N-terminal GTPase (nGTPase) domain of human Miro2 bound to GTP were determined for residues 1–180, confirming that the overall secondary structure closely resembles that of Miro1 nGTPase bound to GTP, with minor variations attributable to crystal packing in the Miro1 structure.","method":"NMR spectroscopy (backbone chemical shift assignments), structural comparison with Miro1 crystal structure","journal":"Biomolecular NMR assignments","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — direct NMR structural characterization of the nGTPase domain, single lab, no functional mutagenesis validation in this paper","pmids":["36050579"],"is_preprint":false},{"year":2018,"finding":"NOS3 inhibition in glial cells preserves Miro-2 levels and prevents axonal mitochondrial fission and restores mitochondrial motility following oxygen-glucose deprivation in white matter, providing post-ischemic protection to both young and aging axons.","method":"Pharmacological NOS3 inhibition, genetic NOS3 deletion, electrophysiology, 3D electron microscopy, Miro-2 level quantification","journal":"The Journal of neuroscience","confidence":"Low","confidence_rationale":"Tier 3 / Weak — Miro-2 preservation is a correlative readout of NOS3 inhibition rather than a direct mechanistic dissection; no direct manipulation of Miro-2 in this study","pmids":["29891729"],"is_preprint":false},{"year":2026,"finding":"Proximity labeling (TurboID) of Miro2 in hippocampal neural stem cells identified CISD1 as a significant interaction partner. Knockdown of both Miro2 and CISD1 impairs mitochondrial trafficking and disrupts stem cell differentiation with increased cytotoxicity. Rescue experiments partially reversed cell death, and both proteins show increased expression and interaction during differentiation.","method":"TurboID proximity labeling, Co-IP, siRNA knockdown, mitochondrial trafficking assays, differentiation assays","journal":"Communications biology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — proximity labeling and single Co-IP for CISD1 interaction, partial rescue only, single lab","pmids":["41663665"],"is_preprint":false},{"year":2026,"finding":"TurboID proximity labeling of Miro2 and its GTP-binding mutant Miro2 T18N showed that abolishing GTP-binding to the N-terminal GTPase domain considerably reduces the number of proteins in proximity of Miro2, demonstrating that the nucleotide state of the N-terminal GTPase domain governs the interaction network and stabilization of Myo19.","method":"TurboID proximity labeling followed by mass spectrometry, GTP-binding mutant (T18N) comparison, cell cycle stage analysis (interphase vs prometaphase)","journal":"Molecular & cellular proteomics : MCP","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — proximity proteomics with GTPase domain mutant directly demonstrates nucleotide-state-dependent interaction network, orthogonal to prior pulldown data","pmids":["42102968"],"is_preprint":false}],"current_model":"RHOT2/Miro2 is an outer mitochondrial membrane Rho GTPase that acts as a multifunctional mitochondrial trafficking adaptor: its N-terminal GTPase domain (structurally characterized by NMR) recruits myosin XIX and coordinates microtubule- and actin-based mitochondrial movement; it undergoes PINK1-mediated phosphorylation (Ser325/Ser430) and Ca2+-EF2-domain-dependent demultimerization to provide a platform for Parkin translocation during mitophagy; it is degraded via Parkin-mediated ubiquitination under hypertrophic stress; it localizes to peroxisomes via its transmembrane domain/Pex19 interaction to negatively regulate Drp1-dependent fission; it promotes mitochondrial nanotunneling and inter-mitochondrial communication in cardiomyocytes; it is sulfhydrated at C185/C504 by CBS/H2S to sustain mitochondrial dynamics and trophoblast motility; it interacts with GCN1 to relay GCN2/ATF4 stress signaling in cancer cells; and it supports tumor invasion through cooperation with MYO9B to suppress RhoA activity, and mediates mitochondrial transfer from cancer cells to fibroblasts to drive CAF differentiation."},"narrative":{"mechanistic_narrative":"RHOT2/Miro2 is an outer mitochondrial membrane Rho-like GTPase that functions as a multifunctional adaptor coordinating mitochondrial positioning, trafficking, and turnover [PMID:30111583, PMID:42102968]. Through its N-terminal GTPase domain it recruits and stabilizes myosin XIX (Myo19), and the nucleotide state of this domain governs both Myo19 association and the broader Miro2 proximity interaction network [PMID:30111583, PMID:42102968]; the GTP-bound nGTPase domain has been characterized structurally by NMR and closely resembles that of Miro1 [PMID:36050579]. Miro2 supports cytoskeleton-based mitochondrial movement and inter-mitochondrial communication, promoting nanotunneling and kissing along microtubules in cardiomyocytes [PMID:31455181], and isoform-specifically driving mitochondrial motility in pancreatic alpha cells to enable glucose-regulated glucagon secretion [PMID:41308986]. During mitochondrial quality control, Miro2 acts as a regulatory platform: PINK1-mediated phosphorylation at Ser325/Ser430 and Ca2+ binding to its EF2 domain drive its demultimerization and realignment, which are required for Parkin translocation to damaged mitochondria, while Parkin-mediated ubiquitination targets Miro2 for proteasomal degradation under hypertrophic stress [PMID:36659543, PMID:31455181]. Miro2 negatively regulates Drp1-dependent fission, acting upstream of Drp1 to preserve mitochondrial integrity, and also localizes to peroxisomes via a Pex19-dependent transmembrane interaction to restrain peroxisomal fission [PMID:33575111, PMID:31894645]. Its function is tuned by CBS/H2S-mediated sulfhydration of cysteines C185 and C504, which sustains mitochondrial dynamics and trophoblast motility [PMID:39461943]. In cancer, Miro2 promotes invasion and metastasis by cooperating with MYO9B to suppress RhoA activity, relays GCN2/ATF4 stress signaling through interaction with GCN1, and mediates mitochondrial transfer from cancer cells to fibroblasts to drive cancer-associated fibroblast differentiation [PMID:39723893, PMID:34992146, PMID:40877413].","teleology":[{"year":2018,"claim":"Establishing how mitochondria engage actin-based motors, Miro2 was identified as a mitochondrial receptor for myosin XIX whose recruitment and Myo19 stability are governed by its N-terminal GTPase domain nucleotide state.","evidence":"Proximity labeling, direct pulldown, and in vivo recruitment/downregulation assays of Miro1/2 and Myo19","pmids":["30111583"],"confidence":"High","gaps":["Did not resolve how GTP loading is regulated on the mitochondrial membrane","Functional consequence of Miro2-specific (vs Miro1) Myo19 recruitment unresolved"]},{"year":2019,"claim":"To define Miro2's role in cardiac mitochondrial communication and turnover, it was shown to promote nanotunneling/kissing and to be degraded by Parkin-mediated ubiquitination under hypertrophic stress.","evidence":"Adenoviral overexpression, transgenic mice, photoactivatable-GFP communication assays, proteasome inhibition, Parkin overexpression","pmids":["31455181"],"confidence":"High","gaps":["Site of Parkin ubiquitination on Miro2 not mapped","Relationship between communication function and degradation not integrated"]},{"year":2019,"claim":"Addressing how Miro2 participates in mitophagy, PINK1 phosphorylation (Ser325/Ser430) and Ca2+ binding at the EF2 domain were shown to drive demultimerization required for Parkin translocation.","evidence":"CCCP treatment, phospho-site and Ca2+-domain mutagenesis, Parkin translocation assays, Miro2 knockout mouse phenotyping","pmids":["36659543"],"confidence":"High","gaps":["Structural basis of tetramer-to-monomer transition not resolved","Whether demultimerization precedes or follows Miro2 degradation unclear"]},{"year":2020,"claim":"Extending Miro localization beyond mitochondria, Miro1/2 were found at peroxisomes where they restrain Drp1-dependent peroxisomal fission via a Pex19-dependent transmembrane interaction.","evidence":"Localization studies, knockdown/knockout with peroxisomal morphology readout, Pex19 interaction assays, domain mapping","pmids":["31894645"],"confidence":"High","gaps":["Partitioning between mitochondria and peroxisomes not quantified","Mechanism of Drp1 suppression at peroxisomes not defined"]},{"year":2020,"claim":"Linking Miro2 loss to mitochondrial dysfunction, knockdown in neural progenitors caused excessive fragmentation and PINK1/Parkin mitophagy, placing Miro2 upstream of Drp1-dependent fission.","evidence":"miR-351-5p targeting, siRNA knockdown with adenoviral rescue, Mdivi-1 epistasis, morphology/function assays","pmids":["33575111"],"confidence":"Medium","gaps":["Direct molecular link between Miro2 and Drp1 not established","Cell-type generality untested"]},{"year":2022,"claim":"To probe Miro2 in cancer signaling, it was shown to bind GCN1 and to be required for GCN2/ATF4 stress-signaling-driven prostate cancer growth.","evidence":"Co-IP, binding-partner network analysis, functional mutagenesis (159L), siRNA, xenografts","pmids":["34992146"],"confidence":"Medium","gaps":["Reciprocal validation and direct binding interface not shown","How a mitochondrial protein couples to GCN1 translational stress signaling unclear"]},{"year":2022,"claim":"Providing a structural foundation, the GTP-bound N-terminal GTPase domain of human Miro2 was assigned by NMR and shown to closely resemble the Miro1 nGTPase fold.","evidence":"NMR backbone chemical shift assignments (residues 1–180) and comparison with Miro1 crystal structure","pmids":["36050579"],"confidence":"Medium","gaps":["No functional mutagenesis in this study","Full-length and EF-hand/second GTPase domain architecture not determined"]},{"year":2024,"claim":"Defining a Miro2-dependent invasion mechanism, Miro2 was shown to cooperate with MYO9B to suppress RhoA, with dual MIRO2/RhoA ablation rescuing invasion.","evidence":"siRNA, double-gene ablation rescue, invasion assays, metastasis models, RhoA activity assays","pmids":["39723893"],"confidence":"Medium","gaps":["Whether Miro2 directly modulates MYO9B GAP activity unknown","Mechanistic link from mitochondrial Miro2 to cytosolic RhoA regulation undefined"]},{"year":2024,"claim":"Identifying redox tuning of Miro2, CBS-derived H2S sulfhydrates C185/C504 to sustain mitochondrial dynamics and trophoblast motility.","evidence":"CBS knockdown, GYY4137 H2S donor, C185S/C504S double mutant, morphology and invasion/migration assays","pmids":["39461943"],"confidence":"Medium","gaps":["Single lab, single study","How sulfhydration alters Miro2 conformation or partner binding not defined"]},{"year":2025,"claim":"Demonstrating tissue-specific motility roles, Miro2 (but not Miro1) was shown to drive mitochondrial motility in pancreatic alpha cells and enable glucose-induced suppression of glucagon secretion.","evidence":"Isoform-selective siRNA, mitochondrial motility imaging, glucagon/insulin secretion and ATP/ADP measurements","pmids":["41308986"],"confidence":"Medium","gaps":["Mechanism of Miro1/Miro2 functional divergence in alpha cells unknown","Single lab, single study"]},{"year":2025,"claim":"Connecting Miro2 to tumor microenvironment remodeling, cancer-cell MIRO2 was shown to be required for mitochondrial transfer to fibroblasts and CAF differentiation.","evidence":"MIRO2 depletion in cancer cells, co-culture and xenograft assays, transfer quantification, CAF marker analysis","pmids":["40877413"],"confidence":"Medium","gaps":["Route of intercellular mitochondrial transfer not defined","Whether Miro2 acts in donor trafficking machinery directly unclear"]},{"year":2026,"claim":"Reinforcing the nucleotide-state control of Miro2's interactome, proximity proteomics with the GTP-binding mutant T18N showed reduced proximal proteins and Myo19 destabilization, and identified CISD1 as a trafficking partner.","evidence":"TurboID proximity labeling with mass spectrometry, T18N mutant comparison, Co-IP, knockdown and differentiation assays","pmids":["42102968","41663665"],"confidence":"Medium","gaps":["CISD1 interaction rests on single Co-IP with partial rescue","Functional contribution of individual proximal proteins not dissected"]},{"year":null,"claim":"How Miro2 mechanistically transmits its outer-membrane adaptor function to cytosolic outputs such as RhoA regulation and GCN1/ATF4 signaling, and how its many post-translational modifications are integrated, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of full-length Miro2 with bound effectors","Integration of phosphorylation, ubiquitination, and sulfhydration codes not mapped","Mechanism coupling mitochondrial Miro2 to cytosolic signaling pathways unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003924","term_label":"GTPase activity","supporting_discovery_ids":[0,10,13]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,13]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[1,5,6]}],"localization":[{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[0,2,3,5]},{"term_id":"GO:0005777","term_label":"peroxisome","supporting_discovery_ids":[1]}],"pathway":[{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[3,5]},{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[1,2,5]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[4,6,9]}],"complexes":[],"partners":["MYO19","PEX19","GCN1","MYO9B","CISD1","PINK1","PARK2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8IXI1","full_name":"Mitochondrial Rho GTPase 2","aliases":["Ras homolog gene family member T2"],"length_aa":618,"mass_kda":68.1,"function":"Atypical mitochondrial nucleoside-triphosphatase (NTPase) involved in mitochondrial trafficking (PubMed:16630562, PubMed:22396657, PubMed:30513825). Probably involved in control of anterograde transport of mitochondria and their subcellular distribution (PubMed:22396657). Can hydrolyze GTP (By similarity). Can hydrolyze ATP and UTP (PubMed:30513825)","subcellular_location":"Mitochondrion outer membrane","url":"https://www.uniprot.org/uniprotkb/Q8IXI1/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RHOT2","classification":"Not Classified","n_dependent_lines":13,"n_total_lines":1208,"dependency_fraction":0.01076158940397351},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"IPO5","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/RHOT2","total_profiled":1310},"omim":[{"mim_id":"613889","title":"RAS HOMOLOG GENE FAMILY, MEMBER T2; RHOT2","url":"https://www.omim.org/entry/613889"},{"mim_id":"613888","title":"RAS HOMOLOG GENE FAMILY, MEMBER T1; RHOT1","url":"https://www.omim.org/entry/613888"},{"mim_id":"300364","title":"ARMADILLO REPEAT-CONTAINING PROTEIN, X-LINKED 3; ARMCX3","url":"https://www.omim.org/entry/300364"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Mitochondria","reliability":"Supported"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/RHOT2"},"hgnc":{"alias_symbol":["MIRO-2","MIRO2"],"prev_symbol":["C16orf39","ARHT2"]},"alphafold":{"accession":"Q8IXI1","domains":[{"cath_id":"3.40.50.300","chopping":"6-169","consensus_level":"high","plddt":91.8129,"start":6,"end":169},{"cath_id":"1.10.238.10","chopping":"181-282","consensus_level":"high","plddt":95.6807,"start":181,"end":282},{"cath_id":"1.10.238.10","chopping":"296-401","consensus_level":"high","plddt":95.7099,"start":296,"end":401},{"cath_id":"3.40.50.300","chopping":"407-583","consensus_level":"high","plddt":84.3548,"start":407,"end":583},{"cath_id":"1.20.5","chopping":"586-618","consensus_level":"medium","plddt":54.4306,"start":586,"end":618}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IXI1","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IXI1-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IXI1-F1-predicted_aligned_error_v6.png","plddt_mean":88.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RHOT2","jax_strain_url":"https://www.jax.org/strain/search?query=RHOT2"},"sequence":{"accession":"Q8IXI1","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8IXI1.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8IXI1/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IXI1"}},"corpus_meta":[{"pmid":"30111583","id":"PMC_30111583","title":"Identification of Miro1 and Miro2 as mitochondrial receptors for myosin XIX.","date":"2018","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/30111583","citation_count":89,"is_preprint":false},{"pmid":"32246085","id":"PMC_32246085","title":"Miro2 tethers the ER to mitochondria to promote mitochondrial fusion in tobacco leaf epidermal cells.","date":"2020","source":"Communications biology","url":"https://pubmed.ncbi.nlm.nih.gov/32246085","citation_count":43,"is_preprint":false},{"pmid":"31894645","id":"PMC_31894645","title":"Peroxisomal fission is modulated by the mitochondrial Rho-GTPases, Miro1 and Miro2.","date":"2020","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/31894645","citation_count":40,"is_preprint":false},{"pmid":"31455181","id":"PMC_31455181","title":"Miro2 Regulates Inter-Mitochondrial Communication in the Heart and Protects Against TAC-Induced Cardiac Dysfunction.","date":"2019","source":"Circulation research","url":"https://pubmed.ncbi.nlm.nih.gov/31455181","citation_count":37,"is_preprint":false},{"pmid":"33575111","id":"PMC_33575111","title":"miR-351-5p/Miro2 axis contributes to hippocampal neural progenitor cell death via unbalanced mitochondrial fission.","date":"2020","source":"Molecular therapy. Nucleic acids","url":"https://pubmed.ncbi.nlm.nih.gov/33575111","citation_count":25,"is_preprint":false},{"pmid":"34992146","id":"PMC_34992146","title":"MIRO2 Regulates Prostate Cancer Cell Growth via GCN1-Dependent Stress Signaling.","date":"2022","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/34992146","citation_count":22,"is_preprint":false},{"pmid":"29891729","id":"PMC_29891729","title":"NOS3 Inhibition Confers Post-Ischemic Protection to Young and Aging White Matter Integrity by Conserving Mitochondrial Dynamics and Miro-2 Levels.","date":"2018","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/29891729","citation_count":18,"is_preprint":false},{"pmid":"40877413","id":"PMC_40877413","title":"MIRO2-mediated mitochondrial transfer from cancer cells induces cancer-associated fibroblast differentiation.","date":"2025","source":"Nature cancer","url":"https://pubmed.ncbi.nlm.nih.gov/40877413","citation_count":13,"is_preprint":false},{"pmid":"22496713","id":"PMC_22496713","title":"Genetic Screening of the Mitochondrial Rho GTPases MIRO1 and MIRO2 in Parkinson's Disease.","date":"2012","source":"The open neurology journal","url":"https://pubmed.ncbi.nlm.nih.gov/22496713","citation_count":10,"is_preprint":false},{"pmid":"39461943","id":"PMC_39461943","title":"Miro2 sulfhydration by CBS/H2S promotes human trophoblast invasion and migration via regulating mitochondria dynamics.","date":"2024","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/39461943","citation_count":9,"is_preprint":false},{"pmid":"36659543","id":"PMC_36659543","title":"Miro2 supplies a platform for Parkin translocation to damaged mitochondria.","date":"2019","source":"Science bulletin","url":"https://pubmed.ncbi.nlm.nih.gov/36659543","citation_count":9,"is_preprint":false},{"pmid":"32948353","id":"PMC_32948353","title":"The role of RHOT1 and RHOT2 genetic variation on Parkinson disease risk and onset.","date":"2020","source":"Neurobiology of aging","url":"https://pubmed.ncbi.nlm.nih.gov/32948353","citation_count":7,"is_preprint":false},{"pmid":"39723893","id":"PMC_39723893","title":"MIRO2 promotes cancer invasion and metastasis via MYO9B suppression of RhoA activity.","date":"2024","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/39723893","citation_count":4,"is_preprint":false},{"pmid":"15218247","id":"PMC_15218247","title":"Cloning and characterization of the mouse Arht2 gene which encodes a putative atypical GTPase.","date":"2004","source":"Cytogenetic and genome research","url":"https://pubmed.ncbi.nlm.nih.gov/15218247","citation_count":4,"is_preprint":false},{"pmid":"36050579","id":"PMC_36050579","title":"NMR resonance assignment of the N-terminal GTPase domain of human Miro2 Bound to GTP.","date":"2022","source":"Biomolecular NMR assignments","url":"https://pubmed.ncbi.nlm.nih.gov/36050579","citation_count":0,"is_preprint":false},{"pmid":"41308986","id":"PMC_41308986","title":"Mitochondrial movement in pancreatic alpha cells requires Miro2 and is regulated by glucose.","date":"2025","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/41308986","citation_count":0,"is_preprint":false},{"pmid":"42102968","id":"PMC_42102968","title":"Dynamic Proximity Networks of Myosin-19 (Myo19) and its Mitochondrial Receptors Miro2 and Metaxin-3.","date":"2026","source":"Molecular & cellular proteomics : MCP","url":"https://pubmed.ncbi.nlm.nih.gov/42102968","citation_count":0,"is_preprint":false},{"pmid":"41663665","id":"PMC_41663665","title":"Proximity labeling unveils potential roles of the Miro2-CISD1 network in mitochondrial dynamics and neuronal differentiation.","date":"2026","source":"Communications biology","url":"https://pubmed.ncbi.nlm.nih.gov/41663665","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.06.16.660032","title":"Comparative Transcriptomics Reveals Inflammatory and Epigenetic Programs that Actively Orchestrate Pineal Brain Sand Calcification","date":"2025-06-20","source":"bioRxiv","url":"https://doi.org/10.1101/2025.06.16.660032","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11174,"output_tokens":3822,"usd":0.045426,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11594,"output_tokens":4227,"usd":0.081823,"stage2_stop_reason":"end_turn"},"total_usd":0.127249,"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\": 2018,\n      \"finding\": \"Miro1 and Miro2 were identified as mitochondrial receptors for myosin XIX (Myo19); Miro1 binds directly to a C-terminal fragment of the Myo19 tail region, and Miro1/2 recruit the Myo19 tail in vivo. This recruitment is regulated by the nucleotide state of the N-terminal Rho-like GTPase domain of Miro1/2. Myo19 protein stability in cells depends on its association with Miro1/2.\",\n      \"method\": \"Proximity labelling, direct interaction/pulldown studies, in vivo recruitment assays, downregulation experiments\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal interaction studies, in vivo recruitment, mechanistic dissection of GTPase domain regulation, replicated across Miro1 and Miro2\",\n      \"pmids\": [\"30111583\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Miro1 and Miro2 localize to peroxisomes (in addition to mitochondria), where they negatively regulate Drp1-dependent peroxisomal fission to maintain peroxisomal size and morphology. Peroxisomal localization of Miro is regulated by its first GTPase domain and mediated by an interaction of its transmembrane domain with the peroxisomal-membrane protein chaperone Pex19.\",\n      \"method\": \"Localization studies, genetic knockdown/knockout with peroxisomal morphology readout, interaction assays (Pex19 binding)\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization experiments with functional readout, identified binding partner (Pex19), domain mapping via GTPase domain and transmembrane domain, multiple orthogonal methods in one study\",\n      \"pmids\": [\"31894645\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Miro2 regulates inter-mitochondrial communication in cardiomyocytes by promoting mitochondrial nanotunneling and kissing along microtubules. Parkin-mediated ubiquitination leads to degradation of Miro2, and proteasome inhibition blocked phenylephrine-induced decrease of Miro2, establishing Parkin as the E3 ubiquitin ligase that targets Miro2 for degradation during hypertrophic stress.\",\n      \"method\": \"Adenovirus-mediated overexpression, transgenic mice, mitochondria-targeted photoactivatable GFP for communication assay, proteasome inhibition, Parkin overexpression\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (live imaging, genetic models, pharmacological inhibition), functional phenotype with defined molecular mechanism (Parkin ubiquitination)\",\n      \"pmids\": [\"31455181\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Miro2 undergoes CCCP-induced demultimerization from a tetramer to a monomer and changes mitochondrial localization (realignment). This realignment is regulated by PINK1-mediated phosphorylation at Ser325/Ser430 and by Ca2+ binding to the EF2 domain, both of which are required for subsequent Parkin translocation to damaged mitochondria during mitophagy.\",\n      \"method\": \"CCCP treatment, phosphorylation site mutagenesis, Ca2+ binding domain analysis, mitophagy/Parkin translocation assays, Miro2 knockout mouse phenotyping\",\n      \"journal\": \"Science bulletin\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — site-specific mutagenesis, defined post-translational modification (PINK1 phosphorylation), Ca2+ binding domain functional validation, in vivo knockout phenotype\",\n      \"pmids\": [\"36659543\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"MIRO2 interacts with General Control Nonderepressible 1 (GCN1) in prostate cancer cells, and this interaction is necessary for efficient GCN1-mediated GCN2 kinase signaling and induction of ATF4. MIRO2 mutation 159L increases GCN1 binding. MIRO2's effect on prostate cancer cell growth is mediated through ATF4.\",\n      \"method\": \"Co-immunoprecipitation, network analysis of binding partners, functional mutagenesis (MIRO2 159L), siRNA knockdown, xenograft tumor models\",\n      \"journal\": \"Molecular cancer research : MCR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP interaction, mutagenesis identifying enhanced binding variant, downstream signaling readout (GCN2/ATF4), single lab\",\n      \"pmids\": [\"34992146\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"miR-351-5p targets Miro2 in hippocampal neural progenitor cells; Miro2 knockdown induces excessive mitochondrial fragmentation, mitochondrial dysfunction (decreased membrane potential, increased ROS), and cell death with subsequent PINK1/Parkin-mediated mitophagy induction. Suppression of mitochondrial fission by Mdivi-1 completely inhibited miR-351-5p-induced cell death, placing Miro2 upstream of Drp1-dependent fission.\",\n      \"method\": \"siRNA knockdown, adenovirus rescue, Mdivi-1 pharmacological inhibition, mitochondrial morphology and function assays\",\n      \"journal\": \"Molecular therapy. Nucleic acids\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with rescue experiment, pharmacological epistasis placing Miro2 upstream of Drp1 fission, multiple functional readouts\",\n      \"pmids\": [\"33575111\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"MIRO2 supports tumor cell invasion and metastasis through cooperation with unconventional myosin 9B (MYO9B); depletion of MIRO2 phenocopies MYO9B depletion (reduced invasion and increased active RhoA), and dual ablation of MIRO2 and RhoA fully rescues tumor cell invasion. MIRO2 is required for MYO9B-driven invasion.\",\n      \"method\": \"siRNA knockdown, double-gene ablation (MIRO2 + RhoA), in vitro invasion assays, mouse metastasis models, RhoA activity assays\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis via double ablation rescue experiment, phenotypic rescue and in vivo model, single lab\",\n      \"pmids\": [\"39723893\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Miro2 cysteines C185 and C504 are sulfhydrated by CBS-derived H2S; this sulfhydration is required for maintenance of mitochondrial dynamics and for trophoblast invasion and migration. Double mutation of Miro2 C185/C504 to serine fragmented mitochondria and inhibited invasion/migration that could not be rescued by H2S donor treatment.\",\n      \"method\": \"CBS knockdown, H2S donor (GYY4137) treatment, site-directed mutagenesis (C185S/C504S double mutant), mitochondrial morphology assays, invasion/migration assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — mutagenesis of specific sulfhydration sites with functional rescue/loss readout, single lab, single study\",\n      \"pmids\": [\"39461943\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Knockdown of Miro2, but not Miro1, reduces mitochondrial motility specifically in pancreatic alpha cells and impairs glucose-induced inhibition of glucagon secretion, without affecting insulin secretion or mitochondrial motility in non-alpha islet cells. Under low glucose, mitochondria are arrested in positions further from the nucleus, correlating with increased ATP/ADP in the sub-plasma membrane space.\",\n      \"method\": \"siRNA knockdown of Miro2 vs Miro1, mitochondrial motility imaging, glucagon/insulin secretion assays, ATP/ADP measurements\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — loss-of-function with specific functional readout and isoform selectivity established, single lab, single study\",\n      \"pmids\": [\"41308986\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MIRO2 in cancer cells is required for mitochondrial transfer to fibroblasts; depletion of MIRO2 in cancer cells suppresses mitochondrial transfer and inhibits cancer-associated fibroblast (CAF) differentiation and tumor growth.\",\n      \"method\": \"MIRO2 depletion in cancer cells, co-culture and xenograft assays, mitochondrial transfer quantification, CAF marker analysis\",\n      \"journal\": \"Nature cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with defined cellular phenotype (blocked mitochondrial transfer), in vitro and in vivo models, single lab\",\n      \"pmids\": [\"40877413\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"NMR backbone resonance assignments of the N-terminal GTPase (nGTPase) domain of human Miro2 bound to GTP were determined for residues 1–180, confirming that the overall secondary structure closely resembles that of Miro1 nGTPase bound to GTP, with minor variations attributable to crystal packing in the Miro1 structure.\",\n      \"method\": \"NMR spectroscopy (backbone chemical shift assignments), structural comparison with Miro1 crystal structure\",\n      \"journal\": \"Biomolecular NMR assignments\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — direct NMR structural characterization of the nGTPase domain, single lab, no functional mutagenesis validation in this paper\",\n      \"pmids\": [\"36050579\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"NOS3 inhibition in glial cells preserves Miro-2 levels and prevents axonal mitochondrial fission and restores mitochondrial motility following oxygen-glucose deprivation in white matter, providing post-ischemic protection to both young and aging axons.\",\n      \"method\": \"Pharmacological NOS3 inhibition, genetic NOS3 deletion, electrophysiology, 3D electron microscopy, Miro-2 level quantification\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — Miro-2 preservation is a correlative readout of NOS3 inhibition rather than a direct mechanistic dissection; no direct manipulation of Miro-2 in this study\",\n      \"pmids\": [\"29891729\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Proximity labeling (TurboID) of Miro2 in hippocampal neural stem cells identified CISD1 as a significant interaction partner. Knockdown of both Miro2 and CISD1 impairs mitochondrial trafficking and disrupts stem cell differentiation with increased cytotoxicity. Rescue experiments partially reversed cell death, and both proteins show increased expression and interaction during differentiation.\",\n      \"method\": \"TurboID proximity labeling, Co-IP, siRNA knockdown, mitochondrial trafficking assays, differentiation assays\",\n      \"journal\": \"Communications biology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — proximity labeling and single Co-IP for CISD1 interaction, partial rescue only, single lab\",\n      \"pmids\": [\"41663665\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"TurboID proximity labeling of Miro2 and its GTP-binding mutant Miro2 T18N showed that abolishing GTP-binding to the N-terminal GTPase domain considerably reduces the number of proteins in proximity of Miro2, demonstrating that the nucleotide state of the N-terminal GTPase domain governs the interaction network and stabilization of Myo19.\",\n      \"method\": \"TurboID proximity labeling followed by mass spectrometry, GTP-binding mutant (T18N) comparison, cell cycle stage analysis (interphase vs prometaphase)\",\n      \"journal\": \"Molecular & cellular proteomics : MCP\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — proximity proteomics with GTPase domain mutant directly demonstrates nucleotide-state-dependent interaction network, orthogonal to prior pulldown data\",\n      \"pmids\": [\"42102968\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RHOT2/Miro2 is an outer mitochondrial membrane Rho GTPase that acts as a multifunctional mitochondrial trafficking adaptor: its N-terminal GTPase domain (structurally characterized by NMR) recruits myosin XIX and coordinates microtubule- and actin-based mitochondrial movement; it undergoes PINK1-mediated phosphorylation (Ser325/Ser430) and Ca2+-EF2-domain-dependent demultimerization to provide a platform for Parkin translocation during mitophagy; it is degraded via Parkin-mediated ubiquitination under hypertrophic stress; it localizes to peroxisomes via its transmembrane domain/Pex19 interaction to negatively regulate Drp1-dependent fission; it promotes mitochondrial nanotunneling and inter-mitochondrial communication in cardiomyocytes; it is sulfhydrated at C185/C504 by CBS/H2S to sustain mitochondrial dynamics and trophoblast motility; it interacts with GCN1 to relay GCN2/ATF4 stress signaling in cancer cells; and it supports tumor invasion through cooperation with MYO9B to suppress RhoA activity, and mediates mitochondrial transfer from cancer cells to fibroblasts to drive CAF differentiation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RHOT2/Miro2 is an outer mitochondrial membrane Rho-like GTPase that functions as a multifunctional adaptor coordinating mitochondrial positioning, trafficking, and turnover [#0, #13]. Through its N-terminal GTPase domain it recruits and stabilizes myosin XIX (Myo19), and the nucleotide state of this domain governs both Myo19 association and the broader Miro2 proximity interaction network [#0, #13]; the GTP-bound nGTPase domain has been characterized structurally by NMR and closely resembles that of Miro1 [#10]. Miro2 supports cytoskeleton-based mitochondrial movement and inter-mitochondrial communication, promoting nanotunneling and kissing along microtubules in cardiomyocytes [#2], and isoform-specifically driving mitochondrial motility in pancreatic alpha cells to enable glucose-regulated glucagon secretion [#8]. During mitochondrial quality control, Miro2 acts as a regulatory platform: PINK1-mediated phosphorylation at Ser325/Ser430 and Ca2+ binding to its EF2 domain drive its demultimerization and realignment, which are required for Parkin translocation to damaged mitochondria, while Parkin-mediated ubiquitination targets Miro2 for proteasomal degradation under hypertrophic stress [#3, #2]. Miro2 negatively regulates Drp1-dependent fission, acting upstream of Drp1 to preserve mitochondrial integrity, and also localizes to peroxisomes via a Pex19-dependent transmembrane interaction to restrain peroxisomal fission [#5, #1]. Its function is tuned by CBS/H2S-mediated sulfhydration of cysteines C185 and C504, which sustains mitochondrial dynamics and trophoblast motility [#7]. In cancer, Miro2 promotes invasion and metastasis by cooperating with MYO9B to suppress RhoA activity, relays GCN2/ATF4 stress signaling through interaction with GCN1, and mediates mitochondrial transfer from cancer cells to fibroblasts to drive cancer-associated fibroblast differentiation [#6, #4, #9].\",\n  \"teleology\": [\n    {\n      \"year\": 2018,\n      \"claim\": \"Establishing how mitochondria engage actin-based motors, Miro2 was identified as a mitochondrial receptor for myosin XIX whose recruitment and Myo19 stability are governed by its N-terminal GTPase domain nucleotide state.\",\n      \"evidence\": \"Proximity labeling, direct pulldown, and in vivo recruitment/downregulation assays of Miro1/2 and Myo19\",\n      \"pmids\": [\"30111583\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve how GTP loading is regulated on the mitochondrial membrane\", \"Functional consequence of Miro2-specific (vs Miro1) Myo19 recruitment unresolved\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"To define Miro2's role in cardiac mitochondrial communication and turnover, it was shown to promote nanotunneling/kissing and to be degraded by Parkin-mediated ubiquitination under hypertrophic stress.\",\n      \"evidence\": \"Adenoviral overexpression, transgenic mice, photoactivatable-GFP communication assays, proteasome inhibition, Parkin overexpression\",\n      \"pmids\": [\"31455181\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Site of Parkin ubiquitination on Miro2 not mapped\", \"Relationship between communication function and degradation not integrated\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Addressing how Miro2 participates in mitophagy, PINK1 phosphorylation (Ser325/Ser430) and Ca2+ binding at the EF2 domain were shown to drive demultimerization required for Parkin translocation.\",\n      \"evidence\": \"CCCP treatment, phospho-site and Ca2+-domain mutagenesis, Parkin translocation assays, Miro2 knockout mouse phenotyping\",\n      \"pmids\": [\"36659543\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of tetramer-to-monomer transition not resolved\", \"Whether demultimerization precedes or follows Miro2 degradation unclear\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Extending Miro localization beyond mitochondria, Miro1/2 were found at peroxisomes where they restrain Drp1-dependent peroxisomal fission via a Pex19-dependent transmembrane interaction.\",\n      \"evidence\": \"Localization studies, knockdown/knockout with peroxisomal morphology readout, Pex19 interaction assays, domain mapping\",\n      \"pmids\": [\"31894645\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Partitioning between mitochondria and peroxisomes not quantified\", \"Mechanism of Drp1 suppression at peroxisomes not defined\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Linking Miro2 loss to mitochondrial dysfunction, knockdown in neural progenitors caused excessive fragmentation and PINK1/Parkin mitophagy, placing Miro2 upstream of Drp1-dependent fission.\",\n      \"evidence\": \"miR-351-5p targeting, siRNA knockdown with adenoviral rescue, Mdivi-1 epistasis, morphology/function assays\",\n      \"pmids\": [\"33575111\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular link between Miro2 and Drp1 not established\", \"Cell-type generality untested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"To probe Miro2 in cancer signaling, it was shown to bind GCN1 and to be required for GCN2/ATF4 stress-signaling-driven prostate cancer growth.\",\n      \"evidence\": \"Co-IP, binding-partner network analysis, functional mutagenesis (159L), siRNA, xenografts\",\n      \"pmids\": [\"34992146\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Reciprocal validation and direct binding interface not shown\", \"How a mitochondrial protein couples to GCN1 translational stress signaling unclear\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Providing a structural foundation, the GTP-bound N-terminal GTPase domain of human Miro2 was assigned by NMR and shown to closely resemble the Miro1 nGTPase fold.\",\n      \"evidence\": \"NMR backbone chemical shift assignments (residues 1–180) and comparison with Miro1 crystal structure\",\n      \"pmids\": [\"36050579\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No functional mutagenesis in this study\", \"Full-length and EF-hand/second GTPase domain architecture not determined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defining a Miro2-dependent invasion mechanism, Miro2 was shown to cooperate with MYO9B to suppress RhoA, with dual MIRO2/RhoA ablation rescuing invasion.\",\n      \"evidence\": \"siRNA, double-gene ablation rescue, invasion assays, metastasis models, RhoA activity assays\",\n      \"pmids\": [\"39723893\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether Miro2 directly modulates MYO9B GAP activity unknown\", \"Mechanistic link from mitochondrial Miro2 to cytosolic RhoA regulation undefined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identifying redox tuning of Miro2, CBS-derived H2S sulfhydrates C185/C504 to sustain mitochondrial dynamics and trophoblast motility.\",\n      \"evidence\": \"CBS knockdown, GYY4137 H2S donor, C185S/C504S double mutant, morphology and invasion/migration assays\",\n      \"pmids\": [\"39461943\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab, single study\", \"How sulfhydration alters Miro2 conformation or partner binding not defined\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Demonstrating tissue-specific motility roles, Miro2 (but not Miro1) was shown to drive mitochondrial motility in pancreatic alpha cells and enable glucose-induced suppression of glucagon secretion.\",\n      \"evidence\": \"Isoform-selective siRNA, mitochondrial motility imaging, glucagon/insulin secretion and ATP/ADP measurements\",\n      \"pmids\": [\"41308986\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of Miro1/Miro2 functional divergence in alpha cells unknown\", \"Single lab, single study\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Connecting Miro2 to tumor microenvironment remodeling, cancer-cell MIRO2 was shown to be required for mitochondrial transfer to fibroblasts and CAF differentiation.\",\n      \"evidence\": \"MIRO2 depletion in cancer cells, co-culture and xenograft assays, transfer quantification, CAF marker analysis\",\n      \"pmids\": [\"40877413\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Route of intercellular mitochondrial transfer not defined\", \"Whether Miro2 acts in donor trafficking machinery directly unclear\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Reinforcing the nucleotide-state control of Miro2's interactome, proximity proteomics with the GTP-binding mutant T18N showed reduced proximal proteins and Myo19 destabilization, and identified CISD1 as a trafficking partner.\",\n      \"evidence\": \"TurboID proximity labeling with mass spectrometry, T18N mutant comparison, Co-IP, knockdown and differentiation assays\",\n      \"pmids\": [\"42102968\", \"41663665\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"CISD1 interaction rests on single Co-IP with partial rescue\", \"Functional contribution of individual proximal proteins not dissected\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How Miro2 mechanistically transmits its outer-membrane adaptor function to cytosolic outputs such as RhoA regulation and GCN1/ATF4 signaling, and how its many post-translational modifications are integrated, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of full-length Miro2 with bound effectors\", \"Integration of phosphorylation, ubiquitination, and sulfhydration codes not mapped\", \"Mechanism coupling mitochondrial Miro2 to cytosolic signaling pathways unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003924\", \"supporting_discovery_ids\": [0, 10, 13]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 13]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [1, 5, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [0, 2, 3, 5]},\n      {\"term_id\": \"GO:0005777\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [3, 5]},\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [1, 2, 5]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [4, 6, 9]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"MYO19\", \"PEX19\", \"GCN1\", \"MYO9B\", \"CISD1\", \"PINK1\", \"PARK2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}