CRISPR screen for protein inclusion formation uncovers a role for SRRD in the regulation of intermediate filament dynamics and aggresome assembly
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Published:2024-02-05
Issue:2
Volume:20
Page:e1011138
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ISSN:1553-7404
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Container-title:PLOS Genetics
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language:en
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Short-container-title:PLoS Genet
Author:
Sweeney Katelyn M.ORCID,
Chantarawong SapannaORCID,
Barbieri Edward M.ORCID,
Cajka GregORCID,
Liu MatthewORCID,
Spruce Lynn,
Fazelinia Hossein,
Portz Bede,
Copley Katie,
Lapidot Tomer,
Duhamel Lauren,
Greenwald PhoebeORCID,
Saida Naseeb,
Shalgi Reut,
Shorter JamesORCID,
Shalem OphirORCID
Abstract
The presence of large protein inclusions is a hallmark of neurodegeneration, and yet the precise molecular factors that contribute to their formation remain poorly understood. Screens using aggregation-prone proteins have commonly relied on downstream toxicity as a readout rather than the direct formation of aggregates. Here, we combined a genome-wide CRISPR knockout screen with Pulse Shape Analysis, a FACS-based method for inclusion detection, to identify direct modifiers of TDP-43 aggregation in human cells. Our screen revealed both canonical and novel proteostasis genes, and unearthed SRRD, a poorly characterized protein, as a top regulator of protein inclusion formation. APEX biotin labeling reveals that SRRD resides in proximity to proteins that are involved in the formation and breakage of disulfide bonds and to intermediate filaments, suggesting a role in regulation of the spatial dynamics of the intermediate filament network. Indeed, loss of SRRD results in aberrant intermediate filament fibrils and the impaired formation of aggresomes, including blunted vimentin cage structure, during proteotoxic stress. Interestingly, SRRD also localizes to aggresomes and unfolded proteins, and rescues proteotoxicity in yeast whereby its N-terminal low complexity domain is sufficient to induce this affect. Altogether this suggests an unanticipated and broad role for SRRD in cytoskeletal organization and cellular proteostasis.
Funder
NIGMS
Pennsylvania Department of Health
NINDS
Sanofi Genzyme
ninds
David and Lucile Packard Foundation
Target ALS
Association for Frontotemporal Degeneration
ALS Association
Office of Disease Prevention and Health Promotion
G. Harold and Leila Y. Mathers Charitable Foundation
nigms
nih
American Health Assistance Foundation
BrightFocus Foundation
Publisher
Public Library of Science (PLoS)
Subject
Cancer Research,Genetics (clinical),Genetics,Molecular Biology,Ecology, Evolution, Behavior and Systematics
Reference103 articles.
1. Differential scales of protein quality control;S Wolff;Cell,2014
2. Cellular strategies of protein quality control;B Chen;Cold Spring Harb Perspect Biol,2011
3. Functional Modules of the Proteostasis Network;GG Jayaraj;Cold Spring Harb Perspect Biol,2020
4. The proteostasis network and its decline in ageing;MS Hipp;Nat Rev Mol Cell Biol,2019
5. Mechanisms and Functions of Spatial Protein Quality Control;EM Sontag;Annu Rev Biochem,2017