{"gene":"DYNC1LI2","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":2009,"finding":"DYNC1LI2 (LIC2) depletion by RNAi causes defects in recycling endosome distribution and cytokinesis, while depletion of DYNC1LI1 (LIC1) does not, demonstrating that LIC2 defines a distinct dynein complex functioning at recycling endosomes versus the LIC1-containing complex at the Golgi.","method":"RNAi knockdown with automated image analysis in mammalian cells","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — RNAi with specific phenotypic readouts (recycling endosome distribution, cytokinesis), replicated across two subunits with reciprocal specificity, reinforced by biochemical stabilization analyses","pmids":["19386764"],"is_preprint":false},{"year":2010,"finding":"DYNC1LI2 (LIC2) RNAi disrupts the distribution of lysosomes and late endosomes; LIC1 and LIC2 are both specifically associated with elements of the late endocytic pathway but not other vesicular compartments, as revealed by isoform-specific antibodies and subcellular fractionation.","method":"RNAi knockdown, isoform-specific antibodies, subcellular localization, RILP-stimulated transport assay","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple complementary approaches (RNAi, antibody-based localization, RILP stimulation rescue), single lab but orthogonal methods","pmids":["21169557"],"is_preprint":false},{"year":2010,"finding":"Rab11-FIP3 binds DYNC1LI2 via the amino-terminal 435 amino acids of FIP3, links Rab11a to DYNC1LI2, recruits DYNC1LI2 onto membranes, and DYNC1LI2 is required for accumulation of endocytosed transferrin at the pericentrosomal endosomal-recycling compartment (ERC).","method":"Co-immunoprecipitation, pulldown, overexpression, endocytosis assay in mammalian cells","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP/pulldown identifying binding partner and domain, supported by functional trafficking assay, single lab","pmids":["20214888"],"is_preprint":false},{"year":2011,"finding":"DYNC1LI2 (LIC2) localizes to spindle poles from prophase through telophase during cell division, while LIC1 localizes to the mitotic spindle and midbody, indicating distinct mitotic roles for LIC1- and LIC2-containing dynein complexes.","method":"Immunofluorescence localization in dividing cells","journal":"Cell biology international","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — direct localization by immunofluorescence at multiple mitotic stages, single lab, no functional perturbation of DYNC1LI2 at spindle poles","pmids":["20964624"],"is_preprint":false},{"year":2021,"finding":"DYNC1LI2 regulates the localization of the chaperone-mediated autophagy (CMA) receptor LAMP2A and CMA activity; reconstitution of DYNC1LI2 in cystinotic (ctns−/−) cells restores endolysosomal dynamics, reduces ER stress, rescues mitochondrial fragmentation, and improves cell survival to oxidative stress. These effects depend on RAB7 and RAB11 activity and LAMP2A, and are not replicated by the paralog DYNC1LI1.","method":"mRNA/protein expression analysis, reconstitution in ctns−/− cells, dominant-negative RAB7/RAB11 expression, LAMP2A knockdown, CMA activity assay, mitochondrial imaging","journal":"Autophagy","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reconstitution in disease model cells with multiple orthogonal functional readouts (CMA, ER stress markers, mitochondria, LAMP2A localization), dominant-negative epistasis, paralog specificity controls, single lab","pmids":["34643468"],"is_preprint":false},{"year":2023,"finding":"KASH5 interacts with DYNC1LI2 (or DYNC1LI1) through a conserved helix in the LIC C-terminal region to act as an activating adaptor for cytoplasmic dynein during meiotic chromosome movement; this region is also required for dynein recruitment to other cellular membranes. LIS1 is essential for dynactin incorporation into the KASH5-dynein complex.","method":"In vitro dynein motility assay, binding domain mapping, mutagenesis of KASH5 EF-hands, co-immunoprecipitation, cell biology assays","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro reconstitution of dynein motility, mutagenesis (EF-hand calcium-binding residues), Co-IP, domain mapping; multiple orthogonal methods in one study","pmids":["36946995"],"is_preprint":false},{"year":2024,"finding":"DYNC1LI2 is specifically methylated at arginine residue 397 by protein arginine methyltransferase 1 (PRMT1), a post-translational modification not found on the paralog DYNC1LI1, identifying a regulatory mechanism specific to LIC2-containing dynein complexes.","method":"Immunoprecipitation, immunoblotting, site-directed mutagenesis, bioinformatics","journal":"Journal of clinical laboratory analysis","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — site-directed mutagenesis identifies specific residue, Co-IP identifies writer enzyme, but single lab and no functional consequence of methylation demonstrated","pmids":["38525916"],"is_preprint":false},{"year":2025,"finding":"Endfoot-localized Dync1li2 mRNA in radial glial cells (RGCs) is critical for RGC basal morphology and interneuron organization; subcellular (endfoot-specific) knockdown of Dync1li2 using CRISPR-Cas13 disrupts RGC basal morphology, demonstrating a compartment-specific function of locally translated DYNC1LI2 in cortical development.","method":"In vivo compartment-specific RNAi (CRISPR-Cas13 LOCAL-KD), RGC compartment purification, subcellular transcriptome analysis","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — novel subcellular knockdown method with defined morphological phenotype in vivo, preprint, single lab, method not yet peer-reviewed","pmids":["41280023"],"is_preprint":true}],"current_model":"DYNC1LI2 (LIC2) is a light intermediate chain subunit of cytoplasmic dynein-1 that defines a functionally distinct dynein complex (separate from LIC1-containing complexes) responsible for recycling endosome distribution, cytokinesis, late endosome/lysosome positioning, LAMP2A-dependent chaperone-mediated autophagy, and spindle pole localization during mitosis; it is recruited to membranes via adaptor proteins including Rab11-FIP3 and KASH5 (through its conserved C-terminal helix), and is post-translationally regulated by PRMT1-mediated arginine-397 methylation."},"narrative":{"mechanistic_narrative":"DYNC1LI2 (LIC2) is a light intermediate chain of cytoplasmic dynein-1 that defines a functionally distinct motor complex from its paralog LIC1, specializing in endomembrane positioning and trafficking [PMID:19386764, PMID:21169557]. Depletion of LIC2, but not LIC1, perturbs recycling endosome distribution and cytokinesis, and disrupts late endosome and lysosome positioning, establishing non-redundant roles for the two LIC-containing dyneins [PMID:19386764, PMID:21169557]. LIC2 is recruited onto membranes through cargo-specific adaptors: Rab11-FIP3 bridges Rab11a to LIC2 to drive accumulation of recycling cargo at the pericentrosomal endosomal-recycling compartment [PMID:20214888], while KASH5 engages a conserved C-terminal helix of the LIC to act as an activating adaptor for dynein motility during meiotic chromosome movement, with this same region serving more broadly in dynein recruitment to membranes [PMID:36946995]. Beyond trafficking, LIC2 regulates localization of the chaperone-mediated autophagy receptor LAMP2A and supports CMA activity, endolysosomal dynamics, and stress resistance in a RAB7/RAB11- and LAMP2A-dependent, paralog-specific manner [PMID:34643468]. LIC2 is post-translationally regulated by PRMT1-mediated methylation at arginine 397, a modification absent from LIC1 [PMID:38525916].","teleology":[{"year":2009,"claim":"Established that LIC2 is not redundant with LIC1 but defines a distinct dynein complex acting at recycling endosomes and in cytokinesis, answering whether the two LIC paralogs are functionally interchangeable.","evidence":"Reciprocal RNAi knockdown of LIC1 and LIC2 with automated phenotypic image analysis in mammalian cells","pmids":["19386764"],"confidence":"High","gaps":["Did not identify the membrane adaptor linking LIC2 to recycling endosomes","No structural basis for paralog-specific cargo selectivity"]},{"year":2010,"claim":"Extended LIC2 function to the late endocytic pathway, showing LIC2 controls lysosome and late endosome distribution alongside LIC1 at these compartments.","evidence":"RNAi, isoform-specific antibody localization, subcellular fractionation, and RILP-stimulated transport assay","pmids":["21169557"],"confidence":"High","gaps":["Mechanism distinguishing LIC1 vs LIC2 contributions at late endosomes not resolved","Adaptor for late-endosome recruitment not defined"]},{"year":2010,"claim":"Identified Rab11-FIP3 as the adaptor coupling Rab11a-positive membranes to LIC2, explaining how LIC2 is recruited to drive recycling-compartment cargo accumulation.","evidence":"Co-immunoprecipitation, pulldown domain mapping (FIP3 N-terminal 435 aa), and transferrin endocytosis assay","pmids":["20214888"],"confidence":"Medium","gaps":["Single lab, no in vitro reconstitution of the FIP3-LIC2-dynein complex","Binding region on LIC2 not mapped"]},{"year":2011,"claim":"Distinguished mitotic localizations of the two LIC complexes, placing LIC2 at spindle poles and LIC1 at spindle/midbody, implying separate mitotic roles.","evidence":"Immunofluorescence at multiple mitotic stages in dividing cells","pmids":["20964624"],"confidence":"Medium","gaps":["No functional perturbation of LIC2 at spindle poles","Mechanism of pole-specific targeting unknown"]},{"year":2021,"claim":"Connected LIC2 to chaperone-mediated autophagy by showing it governs LAMP2A localization and CMA activity, with reconstitution rescuing endolysosomal, ER-stress, and mitochondrial defects in disease-model cells.","evidence":"Reconstitution in ctns-/- cells, dominant-negative RAB7/RAB11, LAMP2A knockdown, CMA assay, mitochondrial imaging","pmids":["34643468"],"confidence":"High","gaps":["Direct LIC2-LAMP2A physical link not established","How RAB7/RAB11 dependence is integrated mechanistically unresolved"]},{"year":2023,"claim":"Defined the LIC C-terminal helix as the binding site for the activating adaptor KASH5 and showed this region also mediates dynein recruitment to other membranes, providing a structural mechanism for LIC2-based motor activation.","evidence":"In vitro dynein motility reconstitution, domain mapping, KASH5 EF-hand mutagenesis, and Co-IP","pmids":["36946995"],"confidence":"High","gaps":["Does not separate LIC1- vs LIC2-specific contributions to KASH5 binding","Regulation of the helix interaction in somatic membrane recruitment unclear"]},{"year":2024,"claim":"Revealed a paralog-specific regulatory layer by identifying PRMT1-mediated arginine-397 methylation on LIC2 but not LIC1.","evidence":"Immunoprecipitation, immunoblotting, site-directed mutagenesis, bioinformatics","pmids":["38525916"],"confidence":"Medium","gaps":["Functional consequence of R397 methylation not demonstrated","Single lab, no in vivo validation"]},{"year":2025,"claim":"Implicated locally translated Dync1li2 mRNA in radial glial endfeet for basal morphology and cortical interneuron organization, extending LIC2 function to compartment-specific roles in development.","evidence":"In vivo CRISPR-Cas13 compartment-specific knockdown, RGC compartment purification, subcellular transcriptome analysis (preprint)","pmids":["41280023"],"confidence":"Medium","gaps":["Preprint, not peer-reviewed","Link between local translation and dynein motor function not mechanistically resolved"]},{"year":null,"claim":"The structural and regulatory logic that makes LIC2 cargo- and process-selective relative to LIC1 remains incompletely defined.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structure of LIC2-containing dynein with paralog-specific adaptors","Functional output of PRMT1 methylation unknown","How a single LIC2 subunit coordinates recycling, CMA, and mitotic roles is unresolved"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[5]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[5]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[2,5]}],"localization":[{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[0,1,2]},{"term_id":"GO:0005764","term_label":"lysosome","supporting_discovery_ids":[1,4]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[3]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[3,5]}],"pathway":[{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[0,1,2]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[4]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,3]}],"complexes":["cytoplasmic dynein-1"],"partners":["RAB11FIP3","KASH5","LIS1","PRMT1","LAMP2A"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O43237","full_name":"Cytoplasmic dynein 1 light intermediate chain 2","aliases":["Dynein light intermediate chain 2, cytosolic","LIC-2","LIC53/55"],"length_aa":492,"mass_kda":54.1,"function":"Acts as one of several non-catalytic accessory components of the cytoplasmic dynein 1 complex that are thought to be involved in linking dynein to cargos and to adapter proteins that regulate dynein function. Cytoplasmic dynein 1 acts as a motor for the intracellular retrograde motility of vesicles and organelles along microtubules. May play a role in binding dynein to membranous organelles or chromosomes","subcellular_location":"Cytoplasm, cytoskeleton","url":"https://www.uniprot.org/uniprotkb/O43237/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/DYNC1LI2","classification":"Not Classified","n_dependent_lines":114,"n_total_lines":1208,"dependency_fraction":0.09437086092715231},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000135720","cell_line_id":"CID001408","localizations":[{"compartment":"cytoplasmic","grade":3},{"compartment":"centrosome","grade":2},{"compartment":"nucleoplasm","grade":2}],"interactors":[{"gene":"DYNC1LI1","stoichiometry":10.0},{"gene":"DYNC1H1","stoichiometry":4.0},{"gene":"DYNC1I2","stoichiometry":4.0},{"gene":"DYNLRB1","stoichiometry":4.0},{"gene":"CAPZB","stoichiometry":0.2},{"gene":"CLIP1","stoichiometry":0.2},{"gene":"CSNK2A2","stoichiometry":0.2},{"gene":"DYNLT1","stoichiometry":0.2},{"gene":"FKBP4","stoichiometry":0.2},{"gene":"ACTR1A","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/target/CID001408","total_profiled":1310},"omim":[{"mim_id":"615890","title":"DYNEIN, CYTOPLASMIC 1, LIGHT INTERMEDIATE CHAIN 1; DYNC1LI1","url":"https://www.omim.org/entry/615890"},{"mim_id":"612319","title":"SPASTIC PARAPLEGIA 35, AUTOSOMAL RECESSIVE, WITH OR WITHOUT NEURODEGENERATION; SPG35","url":"https://www.omim.org/entry/612319"},{"mim_id":"611406","title":"DYNEIN, CYTOPLASMIC 1, LIGHT INTERMEDIATE CHAIN 2; DYNC1LI2","url":"https://www.omim.org/entry/611406"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Centrosome","reliability":"Approved"},{"location":"End piece","reliability":"Approved"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"brain","ntpm":269.4}],"url":"https://www.proteinatlas.org/search/DYNC1LI2"},"hgnc":{"alias_symbol":[],"prev_symbol":["DNCLI2"]},"alphafold":{"accession":"O43237","domains":[{"cath_id":"3.40.50.300","chopping":"55-186_216-338","consensus_level":"high","plddt":74.2295,"start":55,"end":338}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O43237","model_url":"https://alphafold.ebi.ac.uk/files/AF-O43237-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O43237-F1-predicted_aligned_error_v6.png","plddt_mean":61.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=DYNC1LI2","jax_strain_url":"https://www.jax.org/strain/search?query=DYNC1LI2"},"sequence":{"accession":"O43237","fasta_url":"https://rest.uniprot.org/uniprotkb/O43237.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O43237/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O43237"}},"corpus_meta":[{"pmid":"19386764","id":"PMC_19386764","title":"Specificity of cytoplasmic dynein subunits in discrete membrane-trafficking steps.","date":"2009","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/19386764","citation_count":101,"is_preprint":false},{"pmid":"21169557","id":"PMC_21169557","title":"Recruitment of dynein to late endosomes and lysosomes through light intermediate chains.","date":"2010","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/21169557","citation_count":71,"is_preprint":false},{"pmid":"20214888","id":"PMC_20214888","title":"Rab11-FIP3 binds dynein light intermediate chain 2 and its overexpression fragments the Golgi complex.","date":"2010","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/20214888","citation_count":56,"is_preprint":false},{"pmid":"18463364","id":"PMC_18463364","title":"A novel locus for an autosomal recessive hereditary spastic paraplegia (SPG35) maps to 16q21-q23.","date":"2008","source":"Neurology","url":"https://pubmed.ncbi.nlm.nih.gov/18463364","citation_count":43,"is_preprint":false},{"pmid":"23792176","id":"PMC_23792176","title":"Dysferlin interacts with calsequestrin-1, myomesin-2 and dynein in human skeletal muscle.","date":"2013","source":"The international journal of biochemistry & cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/23792176","citation_count":27,"is_preprint":false},{"pmid":"36946995","id":"PMC_36946995","title":"The meiotic LINC complex component KASH5 is an activating adaptor for cytoplasmic dynein.","date":"2023","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/36946995","citation_count":20,"is_preprint":false},{"pmid":"20964624","id":"PMC_20964624","title":"Dynein LIC1 localizes to the mitotic spindle and midbody and LIC2 localizes to spindle poles during cell division.","date":"2011","source":"Cell biology international","url":"https://pubmed.ncbi.nlm.nih.gov/20964624","citation_count":18,"is_preprint":false},{"pmid":"34643468","id":"PMC_34643468","title":"DYNC1LI2 regulates localization of the chaperone-mediated autophagy receptor LAMP2A and improves cellular homeostasis in cystinosis.","date":"2021","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/34643468","citation_count":14,"is_preprint":false},{"pmid":"34645873","id":"PMC_34645873","title":"The expression, localisation and interactome of pigeon CRY2.","date":"2021","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/34645873","citation_count":9,"is_preprint":false},{"pmid":"41280023","id":"PMC_41280023","title":"Subcellular transcriptome of radial glia reveals compartmentalized control of cortical development.","date":"2025","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/41280023","citation_count":2,"is_preprint":false},{"pmid":"35265610","id":"PMC_35265610","title":"Multi-Platform-Based Analysis Characterizes Molecular Alterations of the Nucleus in Human Colorectal Cancer.","date":"2022","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/35265610","citation_count":1,"is_preprint":false},{"pmid":"38525916","id":"PMC_38525916","title":"Dynein Light Intermediate Chains Exhibit Different Arginine Methylation Patterns.","date":"2024","source":"Journal of clinical laboratory analysis","url":"https://pubmed.ncbi.nlm.nih.gov/38525916","citation_count":0,"is_preprint":false},{"pmid":"40683198","id":"PMC_40683198","title":"Impact of copper nanoparticles (CuNPs) on gonadal development in zebrafish larvae and melatonin therapeutic intervention.","date":"2025","source":"Aquatic toxicology (Amsterdam, Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/40683198","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":8572,"output_tokens":2168,"usd":0.029118,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9117,"output_tokens":2986,"usd":0.060117,"stage2_stop_reason":"end_turn"},"total_usd":0.089235,"stage1_batch_id":"msgbatch_01AsR7g8dGTEiEw9U83bJeWn","stage2_batch_id":"msgbatch_01YV5ZyxtFNUZEjxRcpd9jzb","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2009,\n      \"finding\": \"DYNC1LI2 (LIC2) depletion by RNAi causes defects in recycling endosome distribution and cytokinesis, while depletion of DYNC1LI1 (LIC1) does not, demonstrating that LIC2 defines a distinct dynein complex functioning at recycling endosomes versus the LIC1-containing complex at the Golgi.\",\n      \"method\": \"RNAi knockdown with automated image analysis in mammalian cells\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — RNAi with specific phenotypic readouts (recycling endosome distribution, cytokinesis), replicated across two subunits with reciprocal specificity, reinforced by biochemical stabilization analyses\",\n      \"pmids\": [\"19386764\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"DYNC1LI2 (LIC2) RNAi disrupts the distribution of lysosomes and late endosomes; LIC1 and LIC2 are both specifically associated with elements of the late endocytic pathway but not other vesicular compartments, as revealed by isoform-specific antibodies and subcellular fractionation.\",\n      \"method\": \"RNAi knockdown, isoform-specific antibodies, subcellular localization, RILP-stimulated transport assay\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple complementary approaches (RNAi, antibody-based localization, RILP stimulation rescue), single lab but orthogonal methods\",\n      \"pmids\": [\"21169557\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Rab11-FIP3 binds DYNC1LI2 via the amino-terminal 435 amino acids of FIP3, links Rab11a to DYNC1LI2, recruits DYNC1LI2 onto membranes, and DYNC1LI2 is required for accumulation of endocytosed transferrin at the pericentrosomal endosomal-recycling compartment (ERC).\",\n      \"method\": \"Co-immunoprecipitation, pulldown, overexpression, endocytosis assay in mammalian cells\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP/pulldown identifying binding partner and domain, supported by functional trafficking assay, single lab\",\n      \"pmids\": [\"20214888\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"DYNC1LI2 (LIC2) localizes to spindle poles from prophase through telophase during cell division, while LIC1 localizes to the mitotic spindle and midbody, indicating distinct mitotic roles for LIC1- and LIC2-containing dynein complexes.\",\n      \"method\": \"Immunofluorescence localization in dividing cells\",\n      \"journal\": \"Cell biology international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — direct localization by immunofluorescence at multiple mitotic stages, single lab, no functional perturbation of DYNC1LI2 at spindle poles\",\n      \"pmids\": [\"20964624\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"DYNC1LI2 regulates the localization of the chaperone-mediated autophagy (CMA) receptor LAMP2A and CMA activity; reconstitution of DYNC1LI2 in cystinotic (ctns−/−) cells restores endolysosomal dynamics, reduces ER stress, rescues mitochondrial fragmentation, and improves cell survival to oxidative stress. These effects depend on RAB7 and RAB11 activity and LAMP2A, and are not replicated by the paralog DYNC1LI1.\",\n      \"method\": \"mRNA/protein expression analysis, reconstitution in ctns−/− cells, dominant-negative RAB7/RAB11 expression, LAMP2A knockdown, CMA activity assay, mitochondrial imaging\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reconstitution in disease model cells with multiple orthogonal functional readouts (CMA, ER stress markers, mitochondria, LAMP2A localization), dominant-negative epistasis, paralog specificity controls, single lab\",\n      \"pmids\": [\"34643468\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"KASH5 interacts with DYNC1LI2 (or DYNC1LI1) through a conserved helix in the LIC C-terminal region to act as an activating adaptor for cytoplasmic dynein during meiotic chromosome movement; this region is also required for dynein recruitment to other cellular membranes. LIS1 is essential for dynactin incorporation into the KASH5-dynein complex.\",\n      \"method\": \"In vitro dynein motility assay, binding domain mapping, mutagenesis of KASH5 EF-hands, co-immunoprecipitation, cell biology assays\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro reconstitution of dynein motility, mutagenesis (EF-hand calcium-binding residues), Co-IP, domain mapping; multiple orthogonal methods in one study\",\n      \"pmids\": [\"36946995\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"DYNC1LI2 is specifically methylated at arginine residue 397 by protein arginine methyltransferase 1 (PRMT1), a post-translational modification not found on the paralog DYNC1LI1, identifying a regulatory mechanism specific to LIC2-containing dynein complexes.\",\n      \"method\": \"Immunoprecipitation, immunoblotting, site-directed mutagenesis, bioinformatics\",\n      \"journal\": \"Journal of clinical laboratory analysis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — site-directed mutagenesis identifies specific residue, Co-IP identifies writer enzyme, but single lab and no functional consequence of methylation demonstrated\",\n      \"pmids\": [\"38525916\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Endfoot-localized Dync1li2 mRNA in radial glial cells (RGCs) is critical for RGC basal morphology and interneuron organization; subcellular (endfoot-specific) knockdown of Dync1li2 using CRISPR-Cas13 disrupts RGC basal morphology, demonstrating a compartment-specific function of locally translated DYNC1LI2 in cortical development.\",\n      \"method\": \"In vivo compartment-specific RNAi (CRISPR-Cas13 LOCAL-KD), RGC compartment purification, subcellular transcriptome analysis\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — novel subcellular knockdown method with defined morphological phenotype in vivo, preprint, single lab, method not yet peer-reviewed\",\n      \"pmids\": [\"41280023\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"DYNC1LI2 (LIC2) is a light intermediate chain subunit of cytoplasmic dynein-1 that defines a functionally distinct dynein complex (separate from LIC1-containing complexes) responsible for recycling endosome distribution, cytokinesis, late endosome/lysosome positioning, LAMP2A-dependent chaperone-mediated autophagy, and spindle pole localization during mitosis; it is recruited to membranes via adaptor proteins including Rab11-FIP3 and KASH5 (through its conserved C-terminal helix), and is post-translationally regulated by PRMT1-mediated arginine-397 methylation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"DYNC1LI2 (LIC2) is a light intermediate chain of cytoplasmic dynein-1 that defines a functionally distinct motor complex from its paralog LIC1, specializing in endomembrane positioning and trafficking [#0, #1]. Depletion of LIC2, but not LIC1, perturbs recycling endosome distribution and cytokinesis, and disrupts late endosome and lysosome positioning, establishing non-redundant roles for the two LIC-containing dyneins [#0, #1]. LIC2 is recruited onto membranes through cargo-specific adaptors: Rab11-FIP3 bridges Rab11a to LIC2 to drive accumulation of recycling cargo at the pericentrosomal endosomal-recycling compartment [#2], while KASH5 engages a conserved C-terminal helix of the LIC to act as an activating adaptor for dynein motility during meiotic chromosome movement, with this same region serving more broadly in dynein recruitment to membranes [#5]. Beyond trafficking, LIC2 regulates localization of the chaperone-mediated autophagy receptor LAMP2A and supports CMA activity, endolysosomal dynamics, and stress resistance in a RAB7/RAB11- and LAMP2A-dependent, paralog-specific manner [#4]. LIC2 is post-translationally regulated by PRMT1-mediated methylation at arginine 397, a modification absent from LIC1 [#6].\",\n  \"teleology\": [\n    {\n      \"year\": 2009,\n      \"claim\": \"Established that LIC2 is not redundant with LIC1 but defines a distinct dynein complex acting at recycling endosomes and in cytokinesis, answering whether the two LIC paralogs are functionally interchangeable.\",\n      \"evidence\": \"Reciprocal RNAi knockdown of LIC1 and LIC2 with automated phenotypic image analysis in mammalian cells\",\n      \"pmids\": [\"19386764\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not identify the membrane adaptor linking LIC2 to recycling endosomes\", \"No structural basis for paralog-specific cargo selectivity\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Extended LIC2 function to the late endocytic pathway, showing LIC2 controls lysosome and late endosome distribution alongside LIC1 at these compartments.\",\n      \"evidence\": \"RNAi, isoform-specific antibody localization, subcellular fractionation, and RILP-stimulated transport assay\",\n      \"pmids\": [\"21169557\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism distinguishing LIC1 vs LIC2 contributions at late endosomes not resolved\", \"Adaptor for late-endosome recruitment not defined\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Identified Rab11-FIP3 as the adaptor coupling Rab11a-positive membranes to LIC2, explaining how LIC2 is recruited to drive recycling-compartment cargo accumulation.\",\n      \"evidence\": \"Co-immunoprecipitation, pulldown domain mapping (FIP3 N-terminal 435 aa), and transferrin endocytosis assay\",\n      \"pmids\": [\"20214888\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab, no in vitro reconstitution of the FIP3-LIC2-dynein complex\", \"Binding region on LIC2 not mapped\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Distinguished mitotic localizations of the two LIC complexes, placing LIC2 at spindle poles and LIC1 at spindle/midbody, implying separate mitotic roles.\",\n      \"evidence\": \"Immunofluorescence at multiple mitotic stages in dividing cells\",\n      \"pmids\": [\"20964624\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No functional perturbation of LIC2 at spindle poles\", \"Mechanism of pole-specific targeting unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Connected LIC2 to chaperone-mediated autophagy by showing it governs LAMP2A localization and CMA activity, with reconstitution rescuing endolysosomal, ER-stress, and mitochondrial defects in disease-model cells.\",\n      \"evidence\": \"Reconstitution in ctns-/- cells, dominant-negative RAB7/RAB11, LAMP2A knockdown, CMA assay, mitochondrial imaging\",\n      \"pmids\": [\"34643468\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct LIC2-LAMP2A physical link not established\", \"How RAB7/RAB11 dependence is integrated mechanistically unresolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined the LIC C-terminal helix as the binding site for the activating adaptor KASH5 and showed this region also mediates dynein recruitment to other membranes, providing a structural mechanism for LIC2-based motor activation.\",\n      \"evidence\": \"In vitro dynein motility reconstitution, domain mapping, KASH5 EF-hand mutagenesis, and Co-IP\",\n      \"pmids\": [\"36946995\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not separate LIC1- vs LIC2-specific contributions to KASH5 binding\", \"Regulation of the helix interaction in somatic membrane recruitment unclear\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Revealed a paralog-specific regulatory layer by identifying PRMT1-mediated arginine-397 methylation on LIC2 but not LIC1.\",\n      \"evidence\": \"Immunoprecipitation, immunoblotting, site-directed mutagenesis, bioinformatics\",\n      \"pmids\": [\"38525916\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of R397 methylation not demonstrated\", \"Single lab, no in vivo validation\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Implicated locally translated Dync1li2 mRNA in radial glial endfeet for basal morphology and cortical interneuron organization, extending LIC2 function to compartment-specific roles in development.\",\n      \"evidence\": \"In vivo CRISPR-Cas13 compartment-specific knockdown, RGC compartment purification, subcellular transcriptome analysis (preprint)\",\n      \"pmids\": [\"41280023\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint, not peer-reviewed\", \"Link between local translation and dynein motor function not mechanistically resolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural and regulatory logic that makes LIC2 cargo- and process-selective relative to LIC1 remains incompletely defined.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structure of LIC2-containing dynein with paralog-specific adaptors\", \"Functional output of PRMT1 methylation unknown\", \"How a single LIC2 subunit coordinates recycling, CMA, and mitotic roles is unresolved\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [2, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [0, 1, 2]},\n      {\"term_id\": \"GO:0005764\", \"supporting_discovery_ids\": [1, 4]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [3, 5]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [0, 1, 2]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 3]}\n    ],\n    \"complexes\": [\"cytoplasmic dynein-1\"],\n    \"partners\": [\"RAB11FIP3\", \"KASH5\", \"LIS1\", \"PRMT1\", \"LAMP2A\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}