biostudies-literatureByrd ANIA NIH HHSNINDS NIH HHSMichigan Alzheimer's Disease Research CenterNational Institutes of HealthUniversity of ArizonaMichigan Alzheimer’s Disease Research CenterNIH HHSNIGMS NIH HHSNational Science Foundatione202212064https://www.ebi.ac.uk/biostudies/studies/S-EPMC12777955biostudies-literatureUnknown225(2)A pathological hallmark in >97% of amyotrophic lateral sclerosis (ALS) cases is the cytoplasmic mislocalization and aggregation of TDP-43, a nuclear RNA-binding protein, in motor neurons. Driving clearance of cytoplasmic TDP-43 reduces toxicity in ALS models, though how TDP-43 clearance is regulated remains controversial. We conducted an unbiased yeast screen using high-throughput dot blotting to identify genes that affect TDP-43 levels. We identified ESCRT complex genes, which induce membrane invagination (particularly at multivesicular bodies; MVBs) and genes linked to K63 ubiquitination (particularly cofactors of the E3 ubiquitin ligase Rsp5; NEDD4 in humans), as drivers of TDP-43 endolysosomal clearance. TDP-43 colocalized and bound Rsp5/NEDD4 and ESCRT proteins, and perturbations to eThe Journal of cell biologyRsp5/NEDD4 and ESCRT regulate TDP-43 toxicity and turnover via an endolysosomal clearance mechanism.PMC12777955R01GM114564F31NS141379F31 NS134123P330AG072931F31NS134123-01R56 NS128110R56NS128110T32 GM136536DGE-1746060P30 AG072931R01 GM114564R01NS113943F31 NS141379R01 NS113943R01NS097542R01 NS097542T32GM136536Dykstra MMByrd AMarcinowski SAddison VMarmorale LJBuchan JRBarmada SJfalseRsp5/NEDD4 and ESCRT regulate TDP-43 toxicity and turnover via an endolysosomal clearance mechanism.A pathological hallmark in >97% of amyotrophic lateral sclerosis (ALS) cases is the cytoplasmic mislocalization and aggregation of TDP-43, a nuclear RNA-binding protein, in motor neurons. Driving clearance of cytoplasmic TDP-43 reduces toxicity in ALS models, though how TDP-43 clearance is regulated remains controversial. We conducted an unbiased yeast screen using high-throughput dot blotting to identify genes that affect TDP-43 levels. We identified ESCRT complex genes, which induce membrane invagination (particularly at multivesicular bodies; MVBs) and genes linked to K63 ubiquitination (particularly cofactors of the E3 ubiquitin ligase Rsp5; NEDD4 in humans), as drivers of TDP-43 endolysosomal clearance. TDP-43 colocalized and bound Rsp5/NEDD4 and ESCRT proteins, and perturbations to e2026-01-01T00:00:00Z2026 Feb2026-06-06T11:32:55.334Z2026-05-29T03:13:15.454ZS-EPMC127779554149874810.1083/jcb.202212064