Thioredoxin-related protein 32 is an arsenite-regulated thiol reductase of the proteasome 19 S particle

R. Luke Wiseman, King Tung Chin, Cole M. Haynes, Ariel Stanhill, Chong Feng Xu, Assen Roguev, Nevan J. Krogan, Thomas A. Neubert, David Ron

פרסום מחקרי: פרסום בכתב עתמאמרביקורת עמיתים

תקציר

Perturbation of the cytoplasmic protein folding environment by exposure to oxidative stress-inducing As(III)-containing compounds challenges the ubiquitin-proteasome system. Here we report on mass spectrometric analysis of As(III)-induced changes in the proteasome's composition in samples prepared by stable isotope labeling with amino acids in cell culture, using mammalian cells in which TRP32 (thioredoxin-related protein of 32 kDa; also referred to as TXNL1) was identified as a novel subunit of the 26 S proteasome. Quantitative genetic interaction mapping, using the epistatic miniarray profiling approach, identified a functional connection between TRP32 and the proteasome. Deletion of txl1, the Schizosaccharomyces pombe homolog of TRP32, results in a slow growth phenotype when combined with deletion of cut8, a gene required for normal proteasome localization. Deletion analysis in vivo, chemical crosslinking, and manipulation of the ATP concentration in vitro during proteasome immunopurification revealed that the C-terminal domain of mammalian TRP32 binds the 19 S regulatory particle in proximity to the proteasome substrate binding site. Thiol modification with polyethylene glycol-maleimide showed disulfide bond formation at the active site of TRP32 in cells exposed to As(III). Pulse-chase labeling showed that TRP32 is a stable protein whose half-life of >6 h is surprisingly reduced to 1 h upon exposure of cells to As(III). These findings reveal a previously undescribed thiol reductase at the proteasome's regulatory particle.

שפה מקוריתאנגלית
עמודים (מ-עד)15233-15245
מספר עמודים13
כתב עתJournal of Biological Chemistry
כרך284
מספר גיליון22
מזהי עצם דיגיטלי (DOIs)
סטטוס פרסוםפורסם - 29 מאי 2009
פורסם באופן חיצוניכן

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