Genome-wide CRISPR Analysis Identifies Substrate-Specific Conjugation Modules in ER-Associated Degradation

被引：90

作者：

Leto, Dara E. ^{[1
]}

Morgens, David W. ^{[2
]}

Zhang, Lichao ^{[3
]}

Walczak, Christopher P. ^{[1
]}

Elias, Joshua E. ^{[3
]}

Bassik, Michael C. ^{[2
,4
]}

Kopito, Ron R. ^{[1
]}

机构：

[1] Stanford Univ, Dept Biol, Stanford, CA 94305 USA

[2] Stanford Univ, Dept Genet, Stanford, CA 94305 USA

[3] Stanford Univ, Dept Chem & Syst Biol, Stanford, CA 94305 USA

[4] Stanford Univ, Program Chem Engn & Med Human Hlth ChEM H, Stanford, CA 94305 USA

来源：

MOLECULAR CELL | 2019年 / 73卷 / 02期

关键词：

RETICULUM-ASSOCIATED DEGRADATION; SIGNAL PEPTIDE PEPTIDASE; ENDOPLASMIC-RETICULUM; UBIQUITIN-LIGASE; PROTEIN-DEGRADATION; RICIN; PROTEASOME; MECHANISM; CHAINS; CELLS;

D O I：

10.1016/j.molcel.2018.11.015

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The ubiquitin proteasome system (UPS) maintains the integrity of the proteome by selectively degrading misfolded or mis-assembled proteins, but the rules that govern how conformationally defective proteins in the secretory pathway are selected from the structurally and topologically diverse constellation of correctly foldedmembrane and secretory proteins for efficient degradation by cytosolic proteasomes is not well understood. Here, we combine parallel pooled genome-wide CRISPR-Cas9 forward genetic screening with a highly quantitative and sensitive protein turnover assay to discover a previously undescribed collaboration between membrane-embedded cytoplasmic ubiquitin E3 ligases to conjugate heterotypic branched or mixed ubiquitin (Ub) chains on substrates of endoplasmic-reticulumassociated degradation (ERAD). These findings demonstrate that parallel CRISPR analysis can be used to deconvolve highly complex cell biological processes and identify new biochemical pathways in protein quality control.

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页码：377 / +

页数：24

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