Image-based genome-wide siRNA screen identifies selective autophagy factors

Anthony Orvedahl, Rhea Sumpter, Guanghua Xiao, Aylwin Ng, Zhongju Zou, Yi Tang, Masahiro Narimatsu, Christopher Gilpin, Qihua Sun, Michael Roth, Christian V. Forst, Jeffrey L. Wrana, Ying E. Zhang, Katherine Luby-Phelps, Ramnik J. Xavier, Yang Xie, Beth Levine

Research output: Contribution to journalArticle

265 Citations (Scopus)

Abstract

Selective autophagy involves the recognition and targeting of specific cargo, such as damaged organelles, misfolded proteins, or invading pathogens for lysosomal destruction. Yeast genetic screens have identified proteins required for different forms of selective autophagy, including cytoplasm-to-vacuole targeting, pexophagy and mitophagy, and mammalian genetic screens have identified proteins required for autophagy regulation. However, there have been no systematic approaches to identify molecular determinants of selective autophagy in mammalian cells. Here, to identify mammalian genes required for selective autophagy, we performed a high-content, image-based, genome-wide small interfering RNA screen to detect genes required for the colocalization of Sindbis virus capsid protein with autophagolysosomes. We identified 141 candidate genes required for viral autophagy, which were enriched for cellular pathways related to messenger RNA processing, interferon signalling, vesicle trafficking, cytoskeletal motor function and metabolism. Ninety-six of these genes were also required for Parkin-mediated mitophagy, indicating that common molecular determinants may be involved in autophagic targeting of viral nucleocapsids and autophagic targeting of damaged mitochondria. Murine embryonic fibroblasts lacking one of these gene products, the C2-domain containing protein, SMURF1, are deficient in the autophagosomal targeting of Sindbis and herpes simplex viruses and in the clearance of damaged mitochondria. Moreover, SMURF1-deficient mice accumulate damaged mitochondria in the heart, brain and liver. Thus, our study identifies candidate determinants of selective autophagy, and defines SMURF1 as a newly recognized mediator of both viral autophagy and mitophagy.

Original languageEnglish (US)
Pages (from-to)113-117
Number of pages5
JournalNature
Volume480
Issue number7375
DOIs
StatePublished - Dec 1 2011

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Autophagy
Small Interfering RNA
Genome
Mitochondrial Degradation
Genes
Mitochondria
Proteins
Sindbis Virus
Heart Mitochondria
Nucleocapsid
Viral Genes
Liver Mitochondrion
Capsid Proteins
Simplexvirus
Vacuoles
Organelles
Interferons
Cytoplasm
Fibroblasts
Yeasts

ASJC Scopus subject areas

  • General

Cite this

Image-based genome-wide siRNA screen identifies selective autophagy factors. / Orvedahl, Anthony; Sumpter, Rhea; Xiao, Guanghua; Ng, Aylwin; Zou, Zhongju; Tang, Yi; Narimatsu, Masahiro; Gilpin, Christopher; Sun, Qihua; Roth, Michael; Forst, Christian V.; Wrana, Jeffrey L.; Zhang, Ying E.; Luby-Phelps, Katherine; Xavier, Ramnik J.; Xie, Yang; Levine, Beth.

In: Nature, Vol. 480, No. 7375, 01.12.2011, p. 113-117.

Research output: Contribution to journalArticle

Orvedahl, A, Sumpter, R, Xiao, G, Ng, A, Zou, Z, Tang, Y, Narimatsu, M, Gilpin, C, Sun, Q, Roth, M, Forst, CV, Wrana, JL, Zhang, YE, Luby-Phelps, K, Xavier, RJ, Xie, Y & Levine, B 2011, 'Image-based genome-wide siRNA screen identifies selective autophagy factors', Nature, vol. 480, no. 7375, pp. 113-117. https://doi.org/10.1038/nature10546
Orvedahl A, Sumpter R, Xiao G, Ng A, Zou Z, Tang Y et al. Image-based genome-wide siRNA screen identifies selective autophagy factors. Nature. 2011 Dec 1;480(7375):113-117. https://doi.org/10.1038/nature10546
Orvedahl, Anthony ; Sumpter, Rhea ; Xiao, Guanghua ; Ng, Aylwin ; Zou, Zhongju ; Tang, Yi ; Narimatsu, Masahiro ; Gilpin, Christopher ; Sun, Qihua ; Roth, Michael ; Forst, Christian V. ; Wrana, Jeffrey L. ; Zhang, Ying E. ; Luby-Phelps, Katherine ; Xavier, Ramnik J. ; Xie, Yang ; Levine, Beth. / Image-based genome-wide siRNA screen identifies selective autophagy factors. In: Nature. 2011 ; Vol. 480, No. 7375. pp. 113-117.
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