title: The proteasome cap RPT5/Rpt5p subunit prevents aggregation of unfolded ricin A chain.
abstract: The plant cytotoxin ricin enters mammalian cells by receptor-mediated endocytosis, undergoing retrograde transport to the ER (endoplasmic reticulum) where its catalytic A chain (RTA) is reductively separated from the holotoxin to enter the cytosol and inactivate ribosomes. The currently accepted model is that the bulk of ER-dislocated RTA is degraded by proteasomes. We show in the present study that the proteasome has a more complex role in ricin intoxication than previously recognized, that the previously reported increase in sensitivity of mammalian cells to ricin in the presence of proteasome inhibitors simply reflects toxicity of the inhibitors themselves, and that RTA is a very poor substrate for proteasomal degradation. Denatured RTA and casein compete for a binding site on the regulatory particle of the 26S proteasome, but their fates differ. Casein is degraded, but the mammalian 26S proteasome AAA (ATPase associated with various cellular activities)-ATPase subunit RPT5 acts as a chaperone that prevents aggregation of denatured RTA and stimulates recovery of catalytic RTA activity in vitro. Furthermore, in vivo, the ATPase activity of Rpt5p is required for maximal toxicity of RTA dislocated from the Saccharomyces cerevisiae ER. The results of the present study implicate RPT5/Rpt5p in the triage of substrates in which either activation (folding) or inactivation (degradation) pathways may be initiated.
authors.affiliation: School of Life Sciences, University of Warwick, Coventry CV4 7AL, UK.
authors.forename: Paola
authors.initials: P
authors.lastname: Pietroni
authors.forename: Nishi
authors.initials: N
authors.lastname: Vasisht
authors.forename: Jonathan P
authors.initials: JP
authors.lastname: Cook
authors.forename: David M
authors.initials: DM
authors.lastname: Roberts
authors.forename: J Michael
authors.initials: JM
authors.lastname: Lord
authors.forename: Rasmus
authors.initials: R
authors.lastname: Hartmann-Petersen
authors.forename: Lynne M
authors.initials: LM
authors.lastname: Roberts
authors.forename: Robert A
authors.initials: RA
authors.lastname: Spooner
mesh_terms: D000818:Animals; D002364:Caseins / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D012276:Ricin / Q000737:chemistry / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D002364:Caseins; D012276:Ricin; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease