{"gene":"DYNLRB1","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":2006,"finding":"Crystal structure of human DYNLRB1 (Dnlc2A) solved at 2.1 Å resolution by single anomalous diffraction. The protein forms a homodimer with a 10-stranded β-sheet core; residue 89 of each monomer is located within holes in the core and is crucial for binding to the dynein intermediate chain IC74 (DIC). The surface of the β-sheet core is primarily positively charged and predicted to be the site for partner interactions.","method":"X-ray crystallography (SAD), analytical ultracentrifugation/solution studies, mutagenesis inference from structure","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure at 2.1 Å with functional inference from residue-level analysis; single lab but direct structural method","pmids":["16970917"],"is_preprint":false},{"year":2008,"finding":"DYNLRB1 directly interacts with all three Rab6 isoforms (Rab6A, Rab6A', and Rab6B) as shown by yeast two-hybrid, co-immunoprecipitation, and pull-down assays. Pull-down experiments revealed preferred association of DYNLRB1 with GTP-bound Rab6A and GDP-bound Rab6A'/Rab6B. DYNLRB1 co-localizes with EYFP-Rab6 isoforms at the Golgi apparatus. In vitro GTPase activity assays showed DYNLRB1 did not modulate Rab6 GTPase activity.","method":"Yeast two-hybrid, co-immunoprecipitation, pull-down assay, confocal co-localization, in vitro GTPase assay","journal":"Cell motility and the cytoskeleton","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP and pull-down with multiple orthogonal methods (Y2H, Co-IP, pulldown, co-localization) in single lab","pmids":["18044744"],"is_preprint":false},{"year":2009,"finding":"DYNLRB1 interacts directly with the large intracellular loop (between TM6 and TM7) of the human reduced folate carrier (hRFC), as demonstrated by bacterial two-hybrid screen, in vitro pull-down, mammalian two-hybrid luciferase assay, and co-immunoprecipitation. DYNLRB1 co-localizes with hRFC in live HuTu-80 intestinal epithelial cells. Co-expression of DYNLRB1 with hRFC significantly increases folate uptake, while siRNA knockdown of DYNLRB1 or pharmacological inhibition with vanadate significantly reduces folate uptake.","method":"Bacterial two-hybrid, in vitro pull-down, mammalian two-hybrid luciferase assay, co-immunoprecipitation, confocal imaging, siRNA knockdown, functional folate uptake assay","journal":"American journal of physiology. Gastrointestinal and liver physiology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal binding assays plus functional readout in single lab","pmids":["19571232"],"is_preprint":false},{"year":2012,"finding":"DYNLRB1 (km23-1) is required for TGFβ1 autoinduction through Smad2-independent Ras/ERK/JNK pathways. siRNA knockdown of km23-1 reduced TGFβ1 mRNA expression, inhibited TGFβ-mediated ERK and JNK activation, c-Jun phosphorylation, and c-Jun promoter transactivation. Sucrose gradient analysis showed km23-1 co-sediments with Ras and TβRII in lipid rafts after TGFβ treatment. Co-immunoprecipitation demonstrated formation of a TGFβ-inducible complex between Ras and km23-1 within minutes of TGFβ addition. km23-1 is required for Ras activation by TGFβ, functioning as an adaptor coupling TβR activation to Ras effector pathways.","method":"siRNA knockdown, co-immunoprecipitation, sucrose gradient fractionation, reporter assay, immunoblotting","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (Co-IP, sucrose gradient, reporter, siRNA) in single lab","pmids":["22637579"],"is_preprint":false},{"year":2014,"finding":"DYNLRB1 (roadblock-1) is a subunit shared by both cytoplasmic dynein-1 and dynein-2 complexes in human cells. Co-immunoprecipitation and proteomics defined DYNLRB1 as a component of the human dynein-2 complex alongside WDR34, WDR60, TCTEX1D2, DYNLRB2, DYNLT1/DYNLT3, and DYNLL1/DYNLL2.","method":"Co-immunoprecipitation, mass spectrometry proteomics","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and MS proteomics in single lab; multiple subunits confirmed","pmids":["25205765"],"is_preprint":false},{"year":2015,"finding":"DYNLRB1 interacts with N-acetylglucosamine kinase (NAGK) as identified by yeast two-hybrid screening and confirmed by proximity ligation assay and immunocytochemistry in hippocampal neurons. The NAGK-DYNLRB1 interaction localizes to Golgi outposts at dendritic branch points. Introduction of a peptide derived from DYNLRB1 C-terminal residues 74–96 stunted hippocampal neuron dendrites in culture, demonstrating a functional role for this interaction in dendritic growth.","method":"Yeast two-hybrid, proximity ligation assay (PLA), immunocytochemistry, peptide competition assay","journal":"Experimental & molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — PLA and Y2H with functional peptide competition in single lab","pmids":["26272270"],"is_preprint":false},{"year":2015,"finding":"The NAGK-DYNLRB1 (dynein) interaction operates in growing axons where NAGK, dynein (DYNLRB1), and Golgi form a tripartite complex. Overexpression of NAGK increased axonal lengths while shRNA knockdown reduced them; transfection with DYNLRB1(74-96) peptide produced neurons with shorter axons, indicating the NAGK-DYNLRB1 interaction is required for axonal growth.","method":"Proximity ligation assay, immunocytochemistry, overexpression, shRNA knockdown, peptide competition","journal":"Molecules and cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — PLA plus gain/loss-of-function with functional readout; single lab","pmids":["26467288"],"is_preprint":false},{"year":2016,"finding":"DYNLRB1 participates in a complex with NAGK, Lis1, and NudE1 at the nuclear envelope and kinetochores during cell division. PLA showed NAGK-DYNLRB1 complex co-localizes with Lis1 and NudE1 at nuclear envelopes in prophase and on chromosomes in metaphase. shRNA knockdown of NAGK delayed cell division, implicating the NAGK-DYNLRB1-Lis1-NudE1 complex in nuclear envelope breakdown and microtubule-kinetochore attachment.","method":"Proximity ligation assay, immunocytochemistry, shRNA knockdown, cell cycle analysis","journal":"Molecules and cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — PLA plus loss-of-function with defined cell division phenotype; single lab","pmids":["27646688"],"is_preprint":false},{"year":2018,"finding":"Within the dynein-2 complex, DYNLRB1/DYNLRB2 associate with the WDR34 intermediate chain subcomplex (WDR34-DYNLL1/DYNLL2-DYNLRB1/DYNLRB2), as defined by visible immunoprecipitation assay. This places DYNLRB1 in one of three dynein-2 subcomplexes.","method":"Visible immunoprecipitation assay (VIP assay), co-immunoprecipitation","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — VIP assay with defined subcomplex architecture; single lab","pmids":["29742051"],"is_preprint":false},{"year":2019,"finding":"WDR34 intermediate chain binds DYNLRB1/DYNLRB2 via a distinct site separate from its DYNLL1/DYNLL2-binding site. Phenotypic analyses of WDR34-knockout cells expressing WDR34 constructs with disrupted light chain interactions showed that DYNLRB1/DYNLRB2 incorporation into the dynein-2 complex via WDR34 is essential for retrograde ciliary protein trafficking. A WDR34 N-terminal construct encompassing the light chain-binding sites inhibited ciliary biogenesis in a dominant-negative manner.","method":"Co-immunoprecipitation, WDR34-knockout cell lines, exogenous WDR34 construct rescue, dominant-negative expression, ciliary trafficking assay","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Moderate — KO rescue experiments with multiple constructs plus dominant-negative, functional readout; single lab with orthogonal methods","pmids":["30649997"],"is_preprint":false},{"year":2019,"finding":"CRISPR-Cas9 knockout of km23-1/DYNLRB1 reduced cell migration in HCT116 and DLD-1 colorectal cancer cells. Sucrose gradient fractionation showed km23-1/DYNLRB1 co-sediments with Ras, p-ERK, and ERK in fractions that did not contain holo-dynein components, and km23-1/DYNLRB1 co-localizes with R-Ras at the protruding edges of migrating cells. Disruption of dynein motor activity did not reduce TGFβ-mediated MEK1/2 or JNK activation, indicating dynein-independent km23-1/DYNLRB1 functions in Ras/ERK signaling.","method":"CRISPR-Cas9 knockout, siRNA, sucrose gradient fractionation, size exclusion chromatography, immunostaining, cell migration assay","journal":"Cell biology international","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR KO with functional readout plus biochemical fractionation; single lab, multiple methods","pmids":["31393067"],"is_preprint":false},{"year":2020,"finding":"Homozygous Dynlrb1 null mice are embryonic lethal, and heterozygous or adult knockdown animals display reduced neuronal growth. Selective depletion of Dynlrb1 in proprioceptive neurons compromises their survival. Conditional depletion in sensory neurons causes deficits in β-catenin subcellular localization and severe impairment of axonal transport of both lysosomes and retrograde signaling endosomes, establishing DYNLRB1 as essential for general dynein-mediated transport rather than cargo-specific transport.","method":"Knockout mouse (homozygous lethal), conditional KO in sensory neurons, live imaging of axonal transport (lysosome and endosome trafficking), β-catenin localization assay, shRNA knockdown in vivo","journal":"Neurobiology of disease","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vivo genetic KO with multiple phenotypic readouts (lethality, transport deficits, survival), live imaging; single lab but multiple orthogonal approaches","pmids":["32088381"],"is_preprint":false},{"year":2020,"finding":"NAGK interacts with DYNLRB1 and efficiently suppresses mutant huntingtin (Q74) and α-synuclein A53T aggregation in mouse brain cells. Yeast two-hybrid and in silico docking showed the small domain of NAGK (NAGK-DS) binds to the C-terminal of DYNLRB1. A peptide derived from NAGK-DS interfered with Q74 clearance. A kinase-inactive NAGK mutant also cleared aggregates, confirming the effect is structural/non-enzymatic and mediated via DYNLRB1 interaction.","method":"Yeast two-hybrid, protein-protein docking, peptide competition assay, kinase-dead mutant expression, aggregate clearance assay in mouse brain cells","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Y2H, docking, and functional aggregate clearance with kinase-dead control; single lab","pmids":["32796833"],"is_preprint":false},{"year":2020,"finding":"NAGK interacts with NudC and Lis1 within the dynein complex, and these NAGK-NudC-Lis1-dynein complexes (identified via DYNLRB1 interactions) localize to the leading poles of migrating cells. NAGK overexpression accelerated cell migration while shRNA knockdown delayed it; a NAGK peptide from the NudC-interacting domain retarded migration, placing DYNLRB1-containing dynein complexes at the nuclear envelope as regulators of cell migration.","method":"Yeast two-hybrid, pull-down, immunocytochemistry, PLA, wound-healing migration assay, in utero electroporation, peptide competition","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple binding assays plus gain/loss-of-function with migration readout; single lab","pmids":["33374456"],"is_preprint":false},{"year":2023,"finding":"Dynlrb1 is a critical regulator of FMRP function in sensory axons. FMRP associates with endolysosomal organelles (likely through annexin A11) and is retrogradely trafficked by the dynein complex in a Dynlrb1-dependent manner. Dynlrb1 silencing induced FMRP granule accumulation and repressed translation of MAP1B (one of FMRP's primary mRNA targets), establishing Dynlrb1 as required for FMRP retrograde transport and targeted degradation.","method":"Dynlrb1 siRNA silencing, live imaging of retrograde transport, FMRP granule accumulation assay, translational reporter (MAP1B), proteomics","journal":"Molecular & cellular proteomics : MCP","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA loss-of-function with multiple functional readouts (transport, granule accumulation, translation); single lab","pmids":["37739344"],"is_preprint":false},{"year":2025,"finding":"Knockdown of DYNLRB1 (siRNA) mitigated TGFβ ligand-exacerbated α-synuclein-induced toxicity in dopaminergic neurons, placing DYNLRB1 in the TGFβ signaling pathway upstream of α-synuclein toxicity. TGFβ ligand treatment induced upregulation of SNCA mRNA in aSyn-overexpressing cells, and DYNLRB1 knockdown protected against this toxicity similarly to ALK5 and SMAD2 knockdown.","method":"Genome-wide siRNA screen, siRNA knockdown validation, dopaminergic cell death assay, mRNA expression analysis","journal":"Cell death discovery","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA knockdown with cell death functional readout, validated with two independent siRNAs; single lab","pmids":["41387695"],"is_preprint":false}],"current_model":"DYNLRB1 is a homodimeric dynein light chain (roadblock/LC7 family) that is an essential structural subunit shared by both cytoplasmic dynein-1 and dynein-2 complexes, where it associates with the WDR34 intermediate chain subcomplex and is required for general retrograde axonal transport of lysosomes and signaling endosomes, retrograde ciliary protein trafficking via IFT, and FMRP retrograde trafficking/degradation in axons; beyond its motor complex role, DYNLRB1 acts as an adaptor coupling TGFβ receptor activation to Ras/ERK/JNK signaling pathways in a dynein-independent manner, interacts with Rab6 GTPases at the Golgi to regulate retrograde endosome-to-ER transport, and mediates the non-enzymatic structural function of NAGK in axodendritic growth and cell division through a tripartite NAGK-DYNLRB1-Golgi/dynein complex."},"narrative":{"mechanistic_narrative":"DYNLRB1 is a homodimeric roadblock/LC7-family dynein light chain that serves as an essential structural subunit shared by both cytoplasmic dynein-1 and dynein-2 complexes, and additionally functions as a signaling adaptor independent of the motor [PMID:16970917, PMID:25205765, PMID:31393067]. Its 2.1 Å crystal structure reveals a homodimer with a 10-stranded β-sheet core; residue 89 mediates binding to the dynein intermediate chain IC74, and the positively charged β-sheet surface engages partners [PMID:16970917]. Within dynein-2, DYNLRB1 (with DYNLRB2) docks onto the WDR34 intermediate chain via a site distinct from the DYNLL1/DYNLL2 site, and this incorporation is essential for retrograde ciliary protein trafficking by IFT [PMID:29742051, PMID:30649997]. In neurons, DYNLRB1 is required for general dynein-mediated retrograde axonal transport: its loss is embryonic lethal in mice, compromises proprioceptive neuron survival, and impairs transport of both lysosomes and signaling endosomes as well as β-catenin localization [PMID:32088381], and it is specifically required for retrograde trafficking and targeted degradation of FMRP, controlling translation of FMRP targets such as MAP1B [PMID:37739344]. Beyond the motor, DYNLRB1 (km23-1) acts as a dynein-independent adaptor coupling TGFβ receptor activation to Ras/ERK/JNK signaling, co-sedimenting with Ras and TβRII in lipid rafts and forming a TGFβ-inducible Ras complex that drives TGFβ1 autoinduction and cell migration [PMID:22637579, PMID:31393067]. It also binds Rab6 GTPase isoforms at the Golgi in a nucleotide-state-selective manner without altering their GTPase activity [PMID:18044744] and interacts with the reduced folate carrier to promote folate uptake [PMID:19571232]. Through its C-terminal region, DYNLRB1 mediates the non-enzymatic structural function of NAGK in axodendritic growth, mitotic division, and suppression of mutant huntingtin and α-synuclein aggregation [PMID:26272270, PMID:27646688, PMID:32796833].","teleology":[{"year":2006,"claim":"Established the structural basis for DYNLRB1's role as a dynein light chain by defining how it homodimerizes and contacts the intermediate chain.","evidence":"X-ray crystallography (SAD) at 2.1 Å with solution studies and structure-guided mutagenesis inference","pmids":["16970917"],"confidence":"High","gaps":["Functional consequence of residue 89/IC74 binding not tested in cells","Did not address non-dynein partner interfaces"]},{"year":2008,"claim":"Showed DYNLRB1 engages the Golgi-associated Rab6 GTPases in a nucleotide-state-selective manner, implicating it in membrane trafficking beyond classical motor activity.","evidence":"Yeast two-hybrid, co-IP, pull-down, confocal co-localization, in vitro GTPase assay","pmids":["18044744"],"confidence":"High","gaps":["Functional outcome of Rab6 binding on transport not directly demonstrated","Whether binding is dynein-dependent unresolved"]},{"year":2009,"claim":"Linked DYNLRB1 to a transporter, the reduced folate carrier, demonstrating a direct interaction that modulates folate uptake.","evidence":"Bacterial/mammalian two-hybrid, pull-down, co-IP, confocal imaging, siRNA, folate uptake assay","pmids":["19571232"],"confidence":"High","gaps":["Mechanism by which DYNLRB1 enhances hRFC activity (trafficking vs. stabilization) unclear","Physiological relevance in vivo not tested"]},{"year":2012,"claim":"Defined a dynein-independent adaptor role coupling TGFβ receptor activation to Ras/ERK/JNK signaling and TGFβ1 autoinduction.","evidence":"siRNA knockdown, co-IP, sucrose gradient fractionation, reporter assays, immunoblotting","pmids":["22637579"],"confidence":"Medium","gaps":["Direct vs. indirect Ras binding not structurally resolved","Single lab; reciprocal validation of the TGFβ-inducible Ras complex limited"]},{"year":2014,"claim":"Placed DYNLRB1 as a subunit shared by both cytoplasmic dynein-1 and dynein-2, broadening its motor role to ciliary transport.","evidence":"Co-immunoprecipitation and mass spectrometry proteomics","pmids":["25205765"],"confidence":"Medium","gaps":["Stoichiometry within each motor not quantified","Functional necessity within dynein-2 not yet tested here"]},{"year":2015,"claim":"Identified a NAGK-DYNLRB1 interaction at dendritic Golgi outposts and axons that promotes neurite growth via the protein's C-terminal residues.","evidence":"Yeast two-hybrid, PLA, immunocytochemistry, overexpression/shRNA, peptide competition in hippocampal neurons","pmids":["26272270","26467288"],"confidence":"Medium","gaps":["Whether dynein motor activity is required for the growth effect untested","Mechanism linking NAGK binding to Golgi positioning unresolved"]},{"year":2016,"claim":"Extended the NAGK-DYNLRB1 axis to mitosis, placing it with Lis1 and NudE1 at the nuclear envelope and kinetochores during cell division.","evidence":"PLA, immunocytochemistry, shRNA knockdown, cell cycle analysis","pmids":["27646688"],"confidence":"Medium","gaps":["Direct DYNLRB1 contribution (vs. NAGK) to division phenotype not isolated","Mechanism of envelope breakdown role unresolved"]},{"year":2018,"claim":"Resolved the dynein-2 architecture by showing DYNLRB1 binds the WDR34 intermediate chain subcomplex at a site distinct from DYNLL.","evidence":"Visible immunoprecipitation (VIP) assay and co-IP","pmids":["29742051"],"confidence":"Medium","gaps":["Binding sites mapped by IP, not structure","Functional consequence addressed in subsequent work"]},{"year":2019,"claim":"Demonstrated that DYNLRB1 incorporation into dynein-2 via WDR34 is functionally essential for retrograde ciliary protein trafficking.","evidence":"WDR34-knockout cell rescue with interaction-disrupting constructs, dominant-negative expression, ciliary trafficking assay","pmids":["30649997"],"confidence":"High","gaps":["Redundancy between DYNLRB1 and DYNLRB2 not fully dissected","Cargo specificity of ciliary defect unresolved"]},{"year":2019,"claim":"Separated the signaling pool of DYNLRB1 from the motor by showing it co-sediments with Ras/ERK in dynein-free fractions and drives cancer cell migration independently of motor activity.","evidence":"CRISPR-Cas9 knockout, siRNA, sucrose gradient/SEC fractionation, immunostaining, migration assay in colorectal cells","pmids":["31393067"],"confidence":"Medium","gaps":["Molecular composition of the dynein-free signaling complex incomplete","Whether R-Ras binding is direct unresolved"]},{"year":2020,"claim":"Established DYNLRB1 as essential in vivo for general dynein-mediated retrograde axonal transport, with embryonic lethality and broad cargo transport deficits.","evidence":"Constitutive and conditional knockout mice, live imaging of lysosome/endosome transport, β-catenin localization, in vivo shRNA","pmids":["32088381"],"confidence":"High","gaps":["Whether requirement reflects motor assembly or processivity not separated","Tissue-specific dependencies beyond sensory neurons untested"]},{"year":2020,"claim":"Showed the NAGK-DYNLRB1 interaction is non-enzymatic and structural, mediating suppression of huntingtin and α-synuclein aggregation, and operates in migrating cells via Lis1/NudC.","evidence":"Yeast two-hybrid, docking, peptide competition, kinase-dead NAGK control, aggregate clearance and wound-healing assays","pmids":["32796833","33374456"],"confidence":"Medium","gaps":["Mechanism linking aggregate clearance to dynein transport not directly shown","Direct structural interface inferred from docking only"]},{"year":2023,"claim":"Defined DYNLRB1 as required for FMRP retrograde transport and turnover, linking the motor function to local translational control of FMRP targets.","evidence":"siRNA silencing, live retrograde transport imaging, FMRP granule accumulation assay, MAP1B translational reporter, proteomics","pmids":["37739344"],"confidence":"Medium","gaps":["Direct DYNLRB1-FMRP/annexin A11 contacts not biochemically mapped","Mechanism of targeted FMRP degradation unresolved"]},{"year":2025,"claim":"Positioned DYNLRB1 within the TGFβ pathway as a mediator of TGFβ-exacerbated α-synuclein toxicity in dopaminergic neurons.","evidence":"Genome-wide siRNA screen with validation, dopaminergic cell death assay, SNCA mRNA expression analysis","pmids":["41387695"],"confidence":"Medium","gaps":["Whether effect uses the dynein-independent adaptor function not directly tested","Direct molecular target downstream of DYNLRB1 in this pathway unresolved"]},{"year":null,"claim":"It remains unresolved how DYNLRB1 partitions between its motor structural role and its dynein-independent signaling/adaptor functions, and what determines which complex it joins.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of DYNLRB1 in the dynein-independent Ras complex","Regulation of DYNLRB1 partitioning between dynein and signaling pools unknown","Functional overlap/redundancy with DYNLRB2 across contexts undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[0,4,8,9]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3,10]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[2,3]}],"localization":[{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[1,5]},{"term_id":"GO:0005635","term_label":"nuclear envelope","supporting_discovery_ids":[7,13]},{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[9]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[3,10]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[4,11]}],"pathway":[{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[9,11,14]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[3,10,15]},{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[11,14]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[7]}],"complexes":["cytoplasmic dynein-1","cytoplasmic dynein-2 (WDR34 intermediate chain subcomplex)"],"partners":["WDR34","DYNLL1","RAB6A","NAGK","LIS1","NUDE1","RAS","FMRP"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NP97","full_name":"Dynein light chain roadblock-type 1","aliases":["Bithoraxoid-like protein","BLP","Dynein light chain 2A, cytoplasmic","Dynein-associated protein Km23","Roadblock domain-containing protein 1"],"length_aa":96,"mass_kda":10.9,"function":"Component of dynein, a family of motor proteins essential for movement along microtubules (By similarity). Required for structural and functional integrity of cilia (By similarity). 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 (Probable). Cytoplasmic dynein 1 acts as a motor for the intracellular retrograde motility of vesicles and organelles along microtubules (Probable)","subcellular_location":"Cytoplasm, cytoskeleton","url":"https://www.uniprot.org/uniprotkb/Q9NP97/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/DYNLRB1","classification":"Common Essential","n_dependent_lines":1206,"n_total_lines":1208,"dependency_fraction":0.9983443708609272},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"DYNC1H1","stoichiometry":10.0},{"gene":"DYNC1I2","stoichiometry":10.0},{"gene":"DYNC1LI1","stoichiometry":10.0},{"gene":"DYNC2LI1","stoichiometry":10.0},{"gene":"DYNLL2","stoichiometry":10.0},{"gene":"DYNC1LI2","stoichiometry":4.0},{"gene":"DYNLL1","stoichiometry":4.0},{"gene":"CAPZB","stoichiometry":0.2},{"gene":"CLASP1","stoichiometry":0.2},{"gene":"CLIP1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/DYNLRB1","total_profiled":1310},"omim":[{"mim_id":"617353","title":"DYNEIN, LIGHT CHAIN, TCTEX-TYPE, 2B; DYNLT2B","url":"https://www.omim.org/entry/617353"},{"mim_id":"607167","title":"DYNEIN, LIGHT CHAIN, ROADBLOCK TYPE, 1; DYNLRB1","url":"https://www.omim.org/entry/607167"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/DYNLRB1"},"hgnc":{"alias_symbol":["DNLC2A","ROBLD1"],"prev_symbol":["DNCL2A"]},"alphafold":{"accession":"Q9NP97","domains":[{"cath_id":"3.30.450.30","chopping":"16-96","consensus_level":"medium","plddt":95.0386,"start":16,"end":96}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NP97","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NP97-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NP97-F1-predicted_aligned_error_v6.png","plddt_mean":93.19},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=DYNLRB1","jax_strain_url":"https://www.jax.org/strain/search?query=DYNLRB1"},"sequence":{"accession":"Q9NP97","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NP97.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NP97/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NP97"}},"corpus_meta":[{"pmid":"25205765","id":"PMC_25205765","title":"Subunit composition of the human cytoplasmic dynein-2 complex.","date":"2014","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/25205765","citation_count":83,"is_preprint":false},{"pmid":"36343628","id":"PMC_36343628","title":"Tegument protein UL21 of alpha-herpesvirus inhibits the innate immunity by triggering CGAS degradation through TOLLIP-mediated selective autophagy.","date":"2022","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/36343628","citation_count":76,"is_preprint":false},{"pmid":"18044744","id":"PMC_18044744","title":"Rab6 family proteins interact with the dynein light chain protein DYNLRB1.","date":"2008","source":"Cell motility and the cytoskeleton","url":"https://pubmed.ncbi.nlm.nih.gov/18044744","citation_count":64,"is_preprint":false},{"pmid":"11750132","id":"PMC_11750132","title":"Identification of two novel human dynein light chain genes, DNLC2A and DNLC2B, and their expression changes in hepatocellular carcinoma tissues from 68 Chinese patients.","date":"2001","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/11750132","citation_count":56,"is_preprint":false},{"pmid":"29742051","id":"PMC_29742051","title":"Interaction of WDR60 intermediate chain with TCTEX1D2 light chain of the dynein-2 complex is crucial for ciliary protein trafficking.","date":"2018","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/29742051","citation_count":54,"is_preprint":false},{"pmid":"21777384","id":"PMC_21777384","title":"The association of viral proteins with host cell dynein components during virus infection.","date":"2011","source":"The FEBS journal","url":"https://pubmed.ncbi.nlm.nih.gov/21777384","citation_count":51,"is_preprint":false},{"pmid":"30649997","id":"PMC_30649997","title":"Interactions of the dynein-2 intermediate chain WDR34 with the light chains are required for ciliary retrograde protein trafficking.","date":"2019","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/30649997","citation_count":40,"is_preprint":false},{"pmid":"19541378","id":"PMC_19541378","title":"Proteins upregulated by mild and severe hypoxia in squamous cell carcinomas in vitro identified by proteomics.","date":"2009","source":"Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/19541378","citation_count":34,"is_preprint":false},{"pmid":"32088381","id":"PMC_32088381","title":"DYNLRB1 is essential for dynein mediated transport and neuronal survival.","date":"2020","source":"Neurobiology of disease","url":"https://pubmed.ncbi.nlm.nih.gov/32088381","citation_count":23,"is_preprint":false},{"pmid":"19571232","id":"PMC_19571232","title":"Identification of dynein light chain road block-1 as a novel interaction partner with the human reduced folate carrier.","date":"2009","source":"American journal of physiology. 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discovery","url":"https://pubmed.ncbi.nlm.nih.gov/41387695","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":17607,"output_tokens":4820,"usd":0.062561,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":12882,"output_tokens":4604,"usd":0.089755,"stage2_stop_reason":"end_turn"},"total_usd":0.152316,"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\": 2006,\n      \"finding\": \"Crystal structure of human DYNLRB1 (Dnlc2A) solved at 2.1 Å resolution by single anomalous diffraction. The protein forms a homodimer with a 10-stranded β-sheet core; residue 89 of each monomer is located within holes in the core and is crucial for binding to the dynein intermediate chain IC74 (DIC). The surface of the β-sheet core is primarily positively charged and predicted to be the site for partner interactions.\",\n      \"method\": \"X-ray crystallography (SAD), analytical ultracentrifugation/solution studies, mutagenesis inference from structure\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure at 2.1 Å with functional inference from residue-level analysis; single lab but direct structural method\",\n      \"pmids\": [\"16970917\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"DYNLRB1 directly interacts with all three Rab6 isoforms (Rab6A, Rab6A', and Rab6B) as shown by yeast two-hybrid, co-immunoprecipitation, and pull-down assays. Pull-down experiments revealed preferred association of DYNLRB1 with GTP-bound Rab6A and GDP-bound Rab6A'/Rab6B. DYNLRB1 co-localizes with EYFP-Rab6 isoforms at the Golgi apparatus. In vitro GTPase activity assays showed DYNLRB1 did not modulate Rab6 GTPase activity.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, pull-down assay, confocal co-localization, in vitro GTPase assay\",\n      \"journal\": \"Cell motility and the cytoskeleton\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP and pull-down with multiple orthogonal methods (Y2H, Co-IP, pulldown, co-localization) in single lab\",\n      \"pmids\": [\"18044744\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"DYNLRB1 interacts directly with the large intracellular loop (between TM6 and TM7) of the human reduced folate carrier (hRFC), as demonstrated by bacterial two-hybrid screen, in vitro pull-down, mammalian two-hybrid luciferase assay, and co-immunoprecipitation. DYNLRB1 co-localizes with hRFC in live HuTu-80 intestinal epithelial cells. Co-expression of DYNLRB1 with hRFC significantly increases folate uptake, while siRNA knockdown of DYNLRB1 or pharmacological inhibition with vanadate significantly reduces folate uptake.\",\n      \"method\": \"Bacterial two-hybrid, in vitro pull-down, mammalian two-hybrid luciferase assay, co-immunoprecipitation, confocal imaging, siRNA knockdown, functional folate uptake assay\",\n      \"journal\": \"American journal of physiology. Gastrointestinal and liver physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal binding assays plus functional readout in single lab\",\n      \"pmids\": [\"19571232\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"DYNLRB1 (km23-1) is required for TGFβ1 autoinduction through Smad2-independent Ras/ERK/JNK pathways. siRNA knockdown of km23-1 reduced TGFβ1 mRNA expression, inhibited TGFβ-mediated ERK and JNK activation, c-Jun phosphorylation, and c-Jun promoter transactivation. Sucrose gradient analysis showed km23-1 co-sediments with Ras and TβRII in lipid rafts after TGFβ treatment. Co-immunoprecipitation demonstrated formation of a TGFβ-inducible complex between Ras and km23-1 within minutes of TGFβ addition. km23-1 is required for Ras activation by TGFβ, functioning as an adaptor coupling TβR activation to Ras effector pathways.\",\n      \"method\": \"siRNA knockdown, co-immunoprecipitation, sucrose gradient fractionation, reporter assay, immunoblotting\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (Co-IP, sucrose gradient, reporter, siRNA) in single lab\",\n      \"pmids\": [\"22637579\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"DYNLRB1 (roadblock-1) is a subunit shared by both cytoplasmic dynein-1 and dynein-2 complexes in human cells. Co-immunoprecipitation and proteomics defined DYNLRB1 as a component of the human dynein-2 complex alongside WDR34, WDR60, TCTEX1D2, DYNLRB2, DYNLT1/DYNLT3, and DYNLL1/DYNLL2.\",\n      \"method\": \"Co-immunoprecipitation, mass spectrometry proteomics\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and MS proteomics in single lab; multiple subunits confirmed\",\n      \"pmids\": [\"25205765\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"DYNLRB1 interacts with N-acetylglucosamine kinase (NAGK) as identified by yeast two-hybrid screening and confirmed by proximity ligation assay and immunocytochemistry in hippocampal neurons. The NAGK-DYNLRB1 interaction localizes to Golgi outposts at dendritic branch points. Introduction of a peptide derived from DYNLRB1 C-terminal residues 74–96 stunted hippocampal neuron dendrites in culture, demonstrating a functional role for this interaction in dendritic growth.\",\n      \"method\": \"Yeast two-hybrid, proximity ligation assay (PLA), immunocytochemistry, peptide competition assay\",\n      \"journal\": \"Experimental & molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — PLA and Y2H with functional peptide competition in single lab\",\n      \"pmids\": [\"26272270\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"The NAGK-DYNLRB1 (dynein) interaction operates in growing axons where NAGK, dynein (DYNLRB1), and Golgi form a tripartite complex. Overexpression of NAGK increased axonal lengths while shRNA knockdown reduced them; transfection with DYNLRB1(74-96) peptide produced neurons with shorter axons, indicating the NAGK-DYNLRB1 interaction is required for axonal growth.\",\n      \"method\": \"Proximity ligation assay, immunocytochemistry, overexpression, shRNA knockdown, peptide competition\",\n      \"journal\": \"Molecules and cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — PLA plus gain/loss-of-function with functional readout; single lab\",\n      \"pmids\": [\"26467288\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"DYNLRB1 participates in a complex with NAGK, Lis1, and NudE1 at the nuclear envelope and kinetochores during cell division. PLA showed NAGK-DYNLRB1 complex co-localizes with Lis1 and NudE1 at nuclear envelopes in prophase and on chromosomes in metaphase. shRNA knockdown of NAGK delayed cell division, implicating the NAGK-DYNLRB1-Lis1-NudE1 complex in nuclear envelope breakdown and microtubule-kinetochore attachment.\",\n      \"method\": \"Proximity ligation assay, immunocytochemistry, shRNA knockdown, cell cycle analysis\",\n      \"journal\": \"Molecules and cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — PLA plus loss-of-function with defined cell division phenotype; single lab\",\n      \"pmids\": [\"27646688\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Within the dynein-2 complex, DYNLRB1/DYNLRB2 associate with the WDR34 intermediate chain subcomplex (WDR34-DYNLL1/DYNLL2-DYNLRB1/DYNLRB2), as defined by visible immunoprecipitation assay. This places DYNLRB1 in one of three dynein-2 subcomplexes.\",\n      \"method\": \"Visible immunoprecipitation assay (VIP assay), co-immunoprecipitation\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — VIP assay with defined subcomplex architecture; single lab\",\n      \"pmids\": [\"29742051\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"WDR34 intermediate chain binds DYNLRB1/DYNLRB2 via a distinct site separate from its DYNLL1/DYNLL2-binding site. Phenotypic analyses of WDR34-knockout cells expressing WDR34 constructs with disrupted light chain interactions showed that DYNLRB1/DYNLRB2 incorporation into the dynein-2 complex via WDR34 is essential for retrograde ciliary protein trafficking. A WDR34 N-terminal construct encompassing the light chain-binding sites inhibited ciliary biogenesis in a dominant-negative manner.\",\n      \"method\": \"Co-immunoprecipitation, WDR34-knockout cell lines, exogenous WDR34 construct rescue, dominant-negative expression, ciliary trafficking assay\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO rescue experiments with multiple constructs plus dominant-negative, functional readout; single lab with orthogonal methods\",\n      \"pmids\": [\"30649997\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"CRISPR-Cas9 knockout of km23-1/DYNLRB1 reduced cell migration in HCT116 and DLD-1 colorectal cancer cells. Sucrose gradient fractionation showed km23-1/DYNLRB1 co-sediments with Ras, p-ERK, and ERK in fractions that did not contain holo-dynein components, and km23-1/DYNLRB1 co-localizes with R-Ras at the protruding edges of migrating cells. Disruption of dynein motor activity did not reduce TGFβ-mediated MEK1/2 or JNK activation, indicating dynein-independent km23-1/DYNLRB1 functions in Ras/ERK signaling.\",\n      \"method\": \"CRISPR-Cas9 knockout, siRNA, sucrose gradient fractionation, size exclusion chromatography, immunostaining, cell migration assay\",\n      \"journal\": \"Cell biology international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR KO with functional readout plus biochemical fractionation; single lab, multiple methods\",\n      \"pmids\": [\"31393067\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Homozygous Dynlrb1 null mice are embryonic lethal, and heterozygous or adult knockdown animals display reduced neuronal growth. Selective depletion of Dynlrb1 in proprioceptive neurons compromises their survival. Conditional depletion in sensory neurons causes deficits in β-catenin subcellular localization and severe impairment of axonal transport of both lysosomes and retrograde signaling endosomes, establishing DYNLRB1 as essential for general dynein-mediated transport rather than cargo-specific transport.\",\n      \"method\": \"Knockout mouse (homozygous lethal), conditional KO in sensory neurons, live imaging of axonal transport (lysosome and endosome trafficking), β-catenin localization assay, shRNA knockdown in vivo\",\n      \"journal\": \"Neurobiology of disease\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — in vivo genetic KO with multiple phenotypic readouts (lethality, transport deficits, survival), live imaging; single lab but multiple orthogonal approaches\",\n      \"pmids\": [\"32088381\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"NAGK interacts with DYNLRB1 and efficiently suppresses mutant huntingtin (Q74) and α-synuclein A53T aggregation in mouse brain cells. Yeast two-hybrid and in silico docking showed the small domain of NAGK (NAGK-DS) binds to the C-terminal of DYNLRB1. A peptide derived from NAGK-DS interfered with Q74 clearance. A kinase-inactive NAGK mutant also cleared aggregates, confirming the effect is structural/non-enzymatic and mediated via DYNLRB1 interaction.\",\n      \"method\": \"Yeast two-hybrid, protein-protein docking, peptide competition assay, kinase-dead mutant expression, aggregate clearance assay in mouse brain cells\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Y2H, docking, and functional aggregate clearance with kinase-dead control; single lab\",\n      \"pmids\": [\"32796833\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"NAGK interacts with NudC and Lis1 within the dynein complex, and these NAGK-NudC-Lis1-dynein complexes (identified via DYNLRB1 interactions) localize to the leading poles of migrating cells. NAGK overexpression accelerated cell migration while shRNA knockdown delayed it; a NAGK peptide from the NudC-interacting domain retarded migration, placing DYNLRB1-containing dynein complexes at the nuclear envelope as regulators of cell migration.\",\n      \"method\": \"Yeast two-hybrid, pull-down, immunocytochemistry, PLA, wound-healing migration assay, in utero electroporation, peptide competition\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple binding assays plus gain/loss-of-function with migration readout; single lab\",\n      \"pmids\": [\"33374456\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Dynlrb1 is a critical regulator of FMRP function in sensory axons. FMRP associates with endolysosomal organelles (likely through annexin A11) and is retrogradely trafficked by the dynein complex in a Dynlrb1-dependent manner. Dynlrb1 silencing induced FMRP granule accumulation and repressed translation of MAP1B (one of FMRP's primary mRNA targets), establishing Dynlrb1 as required for FMRP retrograde transport and targeted degradation.\",\n      \"method\": \"Dynlrb1 siRNA silencing, live imaging of retrograde transport, FMRP granule accumulation assay, translational reporter (MAP1B), proteomics\",\n      \"journal\": \"Molecular & cellular proteomics : MCP\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA loss-of-function with multiple functional readouts (transport, granule accumulation, translation); single lab\",\n      \"pmids\": [\"37739344\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Knockdown of DYNLRB1 (siRNA) mitigated TGFβ ligand-exacerbated α-synuclein-induced toxicity in dopaminergic neurons, placing DYNLRB1 in the TGFβ signaling pathway upstream of α-synuclein toxicity. TGFβ ligand treatment induced upregulation of SNCA mRNA in aSyn-overexpressing cells, and DYNLRB1 knockdown protected against this toxicity similarly to ALK5 and SMAD2 knockdown.\",\n      \"method\": \"Genome-wide siRNA screen, siRNA knockdown validation, dopaminergic cell death assay, mRNA expression analysis\",\n      \"journal\": \"Cell death discovery\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA knockdown with cell death functional readout, validated with two independent siRNAs; single lab\",\n      \"pmids\": [\"41387695\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"DYNLRB1 is a homodimeric dynein light chain (roadblock/LC7 family) that is an essential structural subunit shared by both cytoplasmic dynein-1 and dynein-2 complexes, where it associates with the WDR34 intermediate chain subcomplex and is required for general retrograde axonal transport of lysosomes and signaling endosomes, retrograde ciliary protein trafficking via IFT, and FMRP retrograde trafficking/degradation in axons; beyond its motor complex role, DYNLRB1 acts as an adaptor coupling TGFβ receptor activation to Ras/ERK/JNK signaling pathways in a dynein-independent manner, interacts with Rab6 GTPases at the Golgi to regulate retrograde endosome-to-ER transport, and mediates the non-enzymatic structural function of NAGK in axodendritic growth and cell division through a tripartite NAGK-DYNLRB1-Golgi/dynein complex.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"DYNLRB1 is a homodimeric roadblock/LC7-family dynein light chain that serves as an essential structural subunit shared by both cytoplasmic dynein-1 and dynein-2 complexes, and additionally functions as a signaling adaptor independent of the motor [#0, #4, #10]. Its 2.1 Å crystal structure reveals a homodimer with a 10-stranded β-sheet core; residue 89 mediates binding to the dynein intermediate chain IC74, and the positively charged β-sheet surface engages partners [#0]. Within dynein-2, DYNLRB1 (with DYNLRB2) docks onto the WDR34 intermediate chain via a site distinct from the DYNLL1/DYNLL2 site, and this incorporation is essential for retrograde ciliary protein trafficking by IFT [#8, #9]. In neurons, DYNLRB1 is required for general dynein-mediated retrograde axonal transport: its loss is embryonic lethal in mice, compromises proprioceptive neuron survival, and impairs transport of both lysosomes and signaling endosomes as well as β-catenin localization [#11], and it is specifically required for retrograde trafficking and targeted degradation of FMRP, controlling translation of FMRP targets such as MAP1B [#14]. Beyond the motor, DYNLRB1 (km23-1) acts as a dynein-independent adaptor coupling TGFβ receptor activation to Ras/ERK/JNK signaling, co-sedimenting with Ras and TβRII in lipid rafts and forming a TGFβ-inducible Ras complex that drives TGFβ1 autoinduction and cell migration [#3, #10]. It also binds Rab6 GTPase isoforms at the Golgi in a nucleotide-state-selective manner without altering their GTPase activity [#1] and interacts with the reduced folate carrier to promote folate uptake [#2]. Through its C-terminal region, DYNLRB1 mediates the non-enzymatic structural function of NAGK in axodendritic growth, mitotic division, and suppression of mutant huntingtin and α-synuclein aggregation [#5, #7, #12].\",\n  \"teleology\": [\n    {\n      \"year\": 2006,\n      \"claim\": \"Established the structural basis for DYNLRB1's role as a dynein light chain by defining how it homodimerizes and contacts the intermediate chain.\",\n      \"evidence\": \"X-ray crystallography (SAD) at 2.1 Å with solution studies and structure-guided mutagenesis inference\",\n      \"pmids\": [\"16970917\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of residue 89/IC74 binding not tested in cells\", \"Did not address non-dynein partner interfaces\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Showed DYNLRB1 engages the Golgi-associated Rab6 GTPases in a nucleotide-state-selective manner, implicating it in membrane trafficking beyond classical motor activity.\",\n      \"evidence\": \"Yeast two-hybrid, co-IP, pull-down, confocal co-localization, in vitro GTPase assay\",\n      \"pmids\": [\"18044744\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional outcome of Rab6 binding on transport not directly demonstrated\", \"Whether binding is dynein-dependent unresolved\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Linked DYNLRB1 to a transporter, the reduced folate carrier, demonstrating a direct interaction that modulates folate uptake.\",\n      \"evidence\": \"Bacterial/mammalian two-hybrid, pull-down, co-IP, confocal imaging, siRNA, folate uptake assay\",\n      \"pmids\": [\"19571232\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which DYNLRB1 enhances hRFC activity (trafficking vs. stabilization) unclear\", \"Physiological relevance in vivo not tested\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Defined a dynein-independent adaptor role coupling TGFβ receptor activation to Ras/ERK/JNK signaling and TGFβ1 autoinduction.\",\n      \"evidence\": \"siRNA knockdown, co-IP, sucrose gradient fractionation, reporter assays, immunoblotting\",\n      \"pmids\": [\"22637579\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs. indirect Ras binding not structurally resolved\", \"Single lab; reciprocal validation of the TGFβ-inducible Ras complex limited\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Placed DYNLRB1 as a subunit shared by both cytoplasmic dynein-1 and dynein-2, broadening its motor role to ciliary transport.\",\n      \"evidence\": \"Co-immunoprecipitation and mass spectrometry proteomics\",\n      \"pmids\": [\"25205765\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Stoichiometry within each motor not quantified\", \"Functional necessity within dynein-2 not yet tested here\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identified a NAGK-DYNLRB1 interaction at dendritic Golgi outposts and axons that promotes neurite growth via the protein's C-terminal residues.\",\n      \"evidence\": \"Yeast two-hybrid, PLA, immunocytochemistry, overexpression/shRNA, peptide competition in hippocampal neurons\",\n      \"pmids\": [\"26272270\", \"26467288\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether dynein motor activity is required for the growth effect untested\", \"Mechanism linking NAGK binding to Golgi positioning unresolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Extended the NAGK-DYNLRB1 axis to mitosis, placing it with Lis1 and NudE1 at the nuclear envelope and kinetochores during cell division.\",\n      \"evidence\": \"PLA, immunocytochemistry, shRNA knockdown, cell cycle analysis\",\n      \"pmids\": [\"27646688\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct DYNLRB1 contribution (vs. NAGK) to division phenotype not isolated\", \"Mechanism of envelope breakdown role unresolved\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Resolved the dynein-2 architecture by showing DYNLRB1 binds the WDR34 intermediate chain subcomplex at a site distinct from DYNLL.\",\n      \"evidence\": \"Visible immunoprecipitation (VIP) assay and co-IP\",\n      \"pmids\": [\"29742051\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Binding sites mapped by IP, not structure\", \"Functional consequence addressed in subsequent work\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Demonstrated that DYNLRB1 incorporation into dynein-2 via WDR34 is functionally essential for retrograde ciliary protein trafficking.\",\n      \"evidence\": \"WDR34-knockout cell rescue with interaction-disrupting constructs, dominant-negative expression, ciliary trafficking assay\",\n      \"pmids\": [\"30649997\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Redundancy between DYNLRB1 and DYNLRB2 not fully dissected\", \"Cargo specificity of ciliary defect unresolved\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Separated the signaling pool of DYNLRB1 from the motor by showing it co-sediments with Ras/ERK in dynein-free fractions and drives cancer cell migration independently of motor activity.\",\n      \"evidence\": \"CRISPR-Cas9 knockout, siRNA, sucrose gradient/SEC fractionation, immunostaining, migration assay in colorectal cells\",\n      \"pmids\": [\"31393067\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular composition of the dynein-free signaling complex incomplete\", \"Whether R-Ras binding is direct unresolved\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Established DYNLRB1 as essential in vivo for general dynein-mediated retrograde axonal transport, with embryonic lethality and broad cargo transport deficits.\",\n      \"evidence\": \"Constitutive and conditional knockout mice, live imaging of lysosome/endosome transport, β-catenin localization, in vivo shRNA\",\n      \"pmids\": [\"32088381\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether requirement reflects motor assembly or processivity not separated\", \"Tissue-specific dependencies beyond sensory neurons untested\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Showed the NAGK-DYNLRB1 interaction is non-enzymatic and structural, mediating suppression of huntingtin and α-synuclein aggregation, and operates in migrating cells via Lis1/NudC.\",\n      \"evidence\": \"Yeast two-hybrid, docking, peptide competition, kinase-dead NAGK control, aggregate clearance and wound-healing assays\",\n      \"pmids\": [\"32796833\", \"33374456\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking aggregate clearance to dynein transport not directly shown\", \"Direct structural interface inferred from docking only\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined DYNLRB1 as required for FMRP retrograde transport and turnover, linking the motor function to local translational control of FMRP targets.\",\n      \"evidence\": \"siRNA silencing, live retrograde transport imaging, FMRP granule accumulation assay, MAP1B translational reporter, proteomics\",\n      \"pmids\": [\"37739344\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct DYNLRB1-FMRP/annexin A11 contacts not biochemically mapped\", \"Mechanism of targeted FMRP degradation unresolved\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Positioned DYNLRB1 within the TGFβ pathway as a mediator of TGFβ-exacerbated α-synuclein toxicity in dopaminergic neurons.\",\n      \"evidence\": \"Genome-wide siRNA screen with validation, dopaminergic cell death assay, SNCA mRNA expression analysis\",\n      \"pmids\": [\"41387695\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether effect uses the dynein-independent adaptor function not directly tested\", \"Direct molecular target downstream of DYNLRB1 in this pathway unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how DYNLRB1 partitions between its motor structural role and its dynein-independent signaling/adaptor functions, and what determines which complex it joins.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of DYNLRB1 in the dynein-independent Ras complex\", \"Regulation of DYNLRB1 partitioning between dynein and signaling pools unknown\", \"Functional overlap/redundancy with DYNLRB2 across contexts undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [0, 4, 8, 9]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3, 10]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [2, 3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [1, 5]},\n      {\"term_id\": \"GO:0005635\", \"supporting_discovery_ids\": [7, 13]},\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [9]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [3, 10]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [4, 11]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [9, 11, 14]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [3, 10, 15]},\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [11, 14]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [7]}\n    ],\n    \"complexes\": [\n      \"cytoplasmic dynein-1\",\n      \"cytoplasmic dynein-2 (WDR34 intermediate chain subcomplex)\"\n    ],\n    \"partners\": [\n      \"WDR34\",\n      \"DYNLL1\",\n      \"Rab6A\",\n      \"NAGK\",\n      \"Lis1\",\n      \"NudE1\",\n      \"Ras\",\n      \"FMRP\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}