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
Affiliations
Michael M. Dubreuil
Program in Cancer Biology, Stanford University, Stanford, CA 94305-5120, USA; Department of Genetics, Stanford University, Stanford, CA 94305-5120, USA
David W. Morgens
Department of Genetics, Stanford University, Stanford, CA 94305-5120, USA
Kanji Okumoto
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
Masanori Honsho
Division of Organelle Homeostasis, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan
Kévin Contrepois
Department of Genetics, Stanford University, Stanford, CA 94305-5120, USA
Brittany Lee-McMullen
Department of Genetics, Stanford University, Stanford, CA 94305-5120, USA
Gavin McAllister Traber
Department of Genetics, Stanford University, Stanford, CA 94305-5120, USA
Ria S. Sood
Department of Genetics, Stanford University, Stanford, CA 94305-5120, USA
Scott J. Dixon
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
Michael P. Snyder
Department of Genetics, Stanford University, Stanford, CA 94305-5120, USA
Yukio Fujiki
Division of Organelle Homeostasis, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan; Corresponding author
Michael C. Bassik
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; Corresponding author
Summary: 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. : Despite its importance in metabolism and disease, a comprehensive analysis of the cellular response to oxidative stress is lacking. Here, Dubreuil et al. use genome-wide screens to identify cellular regulators of oxidative stress. They investigate paradoxical mechanisms by which disruption of the pentose phosphate and peroxisomal import pathways protect cells. Keywords: CRISPR, genome-wide screen, shRNA, pentose phosphate pathway, peroxisomal import pathway, catalase, phosphogluconate dehydrogenase, oxidative stress, reactive oxygen species
