Systematic Identification of Regulators of Oxidative Stress Reveals Non-canonical Roles for Peroxisomal Import and the Pentose Phosphate Pathway

Michael M Dubreuil; David W Morgens; Kanji Okumoto; Masanori Honsho; Kévin Contrepois; Brittany Lee-McMullen; Gavin McAllister Traber; Ria S Sood; Scott J Dixon; Michael P Snyder; Yukio Fujiki; Michael C Bassik

doi:10.1016/j.celrep.2020.01.013

Systematic Identification of Regulators of Oxidative Stress Reveals Non-canonical Roles for Peroxisomal Import and the Pentose Phosphate Pathway

Cell Rep. 2020 Feb 4;30(5):1417-1433.e7. doi: 10.1016/j.celrep.2020.01.013.

Affiliations

¹ Program in Cancer Biology, Stanford University, Stanford, CA 94305-5120, USA; Department of Genetics, Stanford University, Stanford, CA 94305-5120, USA.
² Department of Genetics, Stanford University, Stanford, CA 94305-5120, USA.
³ Department of Biology, Faculty of Sciences, Graduate School of Systems Life Sciences, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan; Division of Organelle Homeostasis, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
⁴ Division of Organelle Homeostasis, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
⁵ Program in Cancer Biology, Stanford University, Stanford, CA 94305-5120, USA; Department of Biology, Stanford University, 327 Campus Drive, Stanford, CA 94305, USA; Chemistry, Engineering, and Medicine for Human Health (ChEM-H), Stanford University, Stanford, CA 94305-5120, USA.
⁶ Division of Organelle Homeostasis, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. Electronic address: yfujiki@kyudai.jp.
⁷ Program in Cancer Biology, Stanford University, Stanford, CA 94305-5120, USA; Department of Genetics, Stanford University, Stanford, CA 94305-5120, USA; Chemistry, Engineering, and Medicine for Human Health (ChEM-H), Stanford University, Stanford, CA 94305-5120, USA. Electronic address: bassik@stanford.edu.

Abstract

Reactive oxygen species (ROS) play critical roles in metabolism and disease, yet a comprehensive analysis of the cellular response to oxidative stress is lacking. To systematically identify regulators of oxidative stress, we conducted genome-wide Cas9/CRISPR and shRNA screens. This revealed a detailed picture of diverse pathways that control oxidative stress response, ranging from the TCA cycle and DNA repair machineries to iron transport, trafficking, and metabolism. Paradoxically, disrupting the pentose phosphate pathway (PPP) at the level of phosphogluconate dehydrogenase (PGD) protects cells against ROS. This dramatically alters metabolites in the PPP, consistent with rewiring of upper glycolysis to promote antioxidant production. In addition, disruption of peroxisomal import unexpectedly increases resistance to oxidative stress by altering the localization of catalase. Together, these studies provide insights into the roles of peroxisomal matrix import and the PPP in redox biology and represent a rich resource for understanding the cellular response to oxidative stress.

Keywords: CRISPR; catalase; genome-wide screen; oxidative stress; pentose phosphate pathway; peroxisomal import pathway; phosphogluconate dehydrogenase; reactive oxygen species; shRNA.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems
Catalase / metabolism
Cytoprotection
Cytosol / metabolism
Genome, Human
Glucose / metabolism
Glycolysis
HeLa Cells
Humans
K562 Cells
Oxidative Stress*
Pentose Phosphate Pathway*
Peroxisomes / metabolism*
Phosphogluconate Dehydrogenase
Protein Transport
RNA, Small Interfering / metabolism
Reactive Oxygen Species / metabolism

Substances

RNA, Small Interfering
Reactive Oxygen Species
Phosphogluconate Dehydrogenase
Catalase
Glucose

Systematic Identification of Regulators of Oxidative Stress Reveals Non-canonical Roles for Peroxisomal Import and the Pentose Phosphate Pathway

Authors

Affiliations

Abstract

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MeSH terms

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