title: The ubiquitin isopeptidase UBPY regulates endosomal ubiquitin dynamics and is essential for receptor down-regulation.
abstract: UBPY is a ubiquitin-specific protease that can deubiquitinate monoubiquitinated receptor tyrosine kinases, as well as process Lys-48- and Lys-63-linked polyubiquitin to lower denomination forms in vitro. Catalytically inactive UBPY localizes to endosomes, which accumulate ubiquitinated proteins. We have explored the sequelae of short interfering RNA-mediated knockdown of UBPY. Global levels of ubiquitinated protein increase and ubiquitin accumulates on endosomes, although free ubiquitin levels are unchanged. UBPY-depleted cells have more and larger multivesicular endosomal structures that are frequently associated through extended contact areas, characterized by regularly spaced, electron-dense, bridging profiles. Degradation of acutely stimulated receptor tyrosine kinases, epidermal growth factor receptor and Met, is strongly inhibited in UBPY knockdown cells suggesting that UBPY function is essential for growth factor receptor down-regulation. In contrast, stability of the UBPY binding partner STAM is dramatically compromised in UBPY knockdown cells. The cellular functions of UBPY are complex but clearly distinct from those of the Lys-63-ubiquitin-specific protease, AMSH, with which it shares a binding site on the SH3 domain of STAM.
authors.affiliation: Physiological Laboratory, University of Liverpool, Crown Street, L69 3BX Liverpool, United Kingdom.
authors.forename: Paula E
authors.initials: PE
authors.lastname: Row
authors.forename: Ian A
authors.initials: IA
authors.lastname: Prior
authors.forename: John
authors.initials: J
authors.lastname: McCullough
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Clague
authors.forename: Sylvie
authors.initials: S
authors.lastname: Urbé
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry; D015536:Down-Regulation; D010450:Endopeptidases / Q000737:chemistry* / Q000502:physiology*; D056827:Endosomal Sorting Complexes Required for Transport; D011992:Endosomes / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008239:Lysine / Q000737:chemistry; D009154:Mutation; D010750:Phosphoproteins / Q000737:chemistry; D012313:RNA / Q000737:chemistry; D025801:Ubiquitin / Q000737:chemistry* / Q000378:metabolism; D043222:Ubiquitin Thiolesterase; D018909:src Homology Domains
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D056827:Endosomal Sorting Complexes Required for Transport; D010750:Phosphoproteins; C109775:STAM protein, human; C486696:STAMBP protein, human; D025801:Ubiquitin; D049452:Green Fluorescent Proteins; D012313:RNA; D010450:Endopeptidases; C541534:USP8 protein, human; D043222:Ubiquitin Thiolesterase; C091939:ubiquitin isopeptidase; D008239:Lysine