biostudies-literatureAdoff HNIGMS NIH HHSNIH HHSar114https://www.ebi.ac.uk/biostudies/studies/S-EPMC12404652biostudies-literatureUnknown36(9)Endosomes are a central sorting hub for membrane cargos. DNAJC13/RME-8 plays a critical role in endosomal trafficking by regulating the endosomal recycling and degradative pathways. DNAJC13 localizes to endosomes through its N-terminal Pleckstrin Homology (PH)-like domain, which binds endosomal phosphatidylinositol-3-phosphate (PI(3)P). However, little is known about how DNAJC13 localization is regulated. Here, we show that two regions within DNAJC13, its J domain and disordered C-terminus, act as negative regulators of its PH-like domain. Using a structure-function approach, we map these control points to a conserved YLT motif in the disordered C-terminus as well as the catalytic HPD triad in its J domain. Mutation of either motif enhances DNAJC13 endosomal localization in cells and increMolecular biology of the cellDNAJC13 localization to endosomes is opposed by its J domain and its disordered C-terminus.PMC12404652S10 OD034224R35 GM137835T32 GM142619Adoff HLobingier BTNovy BHolland EfalseDNAJC13 localization to endosomes is opposed by its J domain and its disordered C-terminus.Endosomes are a central sorting hub for membrane cargos. DNAJC13/RME-8 plays a critical role in endosomal trafficking by regulating the endosomal recycling and degradative pathways. DNAJC13 localizes to endosomes through its N-terminal Pleckstrin Homology (PH)-like domain, which binds endosomal phosphatidylinositol-3-phosphate (PI(3)P). However, little is known about how DNAJC13 localization is regulated. Here, we show that two regions within DNAJC13, its J domain and disordered C-terminus, act as negative regulators of its PH-like domain. Using a structure-function approach, we map these control points to a conserved YLT motif in the disordered C-terminus as well as the catalytic HPD triad in its J domain. Mutation of either motif enhances DNAJC13 endosomal localization in cells and incre2025-01-01T00:00:00Z2025 Sep2026-06-03T06:06:47.504Z2026-04-25T03:21:17.38ZS-EPMC124046524073728610.1091/mbc.E24-12-0575