A Genetic Map of the Response to DNA Damage in Human Cells.

Abstract

The response to DNA damage is critical for cellular homeostasis, tumor suppression, immunity, and gametogenesis. In order to provide an unbiased and global view of the DNA damage response in human cells, we undertook 31 CRISPR-Cas9 screens against 27 genotoxic agents in the retinal pigment epithelium-1 (RPE1) cell line. These screens identified 890 genes whose loss causes either sensitivity or resistance to DNA-damaging agents. Mining this dataset, we discovered that ERCC6L2 (which is mutated in a bone-marrow failure syndrome) codes for a canonical non-homologous end-joining pathway factor, that the RNA polymerase II component ELOF1 modulates the response to transcription-blocking agents, and that the cytotoxicity of the G-quadruplex ligand pyridostatin involves trapping topoisomerase II on DNA. This map of the DNA damage response provides a rich resource to study this fundamental cellular system and has implications for the development and use of genotoxic agents in cancer therapy.

• Authors: Michele Olivieri, Tiffany Cho, Alejandro Álvarez-Quilón, Kejiao Li, Matthew J Schellenberg, Michal Zimmermann, Nicole Hustedt, Silvia Emma Rossi, Salomé Adam, Henrique Melo, Anne Margriet Heijink, Guillermo Sastre-Moreno, Nathalie Moatti, Rachel K Szilard, Andrea McEwan, Alexanda K Ling, Almudena Serrano-Benitez, Tajinder Ubhi, Sumin Feng, Judy Pawling, Irene Delgado-Sainz, Michael W Ferguson, James W Dennis, Grant W Brown, Felipe Cortés-Ledesma, R Scott Williams, Alberto Martin, Dongyi Xu, Daniel Durocher
• Journal: Cell (Cell) Vol. 182 Issue 2 Pg. 481-496.e21 (07 23 2020) ISSN: 1097-4172 [Electronic] United States
• PMID: 32649862 (Publication Type: Journal Article, Research Support, N.I.H., Intramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2020 Elsevier Inc. All rights reserved.

Chemical References

• Aminoquinolines
• Picolinic Acids
• RNA, Guide
• Tumor Suppressor Protein p53
• pyridostatin
• Cytochrome-B(5) Reductase
• DNA Helicases
• ERCC6L2 protein, human
• DNA Topoisomerases, Type II

Topics

• Aminoquinolines (pharmacology)
• Animals
• CRISPR-Cas Systems (genetics)
• Cell Line
• Cytochrome-B(5) Reductase (genetics, metabolism)
• DNA Damage (drug effects)
• DNA Helicases (genetics, metabolism)
• DNA Repair
• DNA Topoisomerases, Type II (genetics, metabolism)
• Gene Regulatory Networks (physiology)
• Humans
• Mice
• Picolinic Acids (pharmacology)
• RNA, Guide, Kinetoplastida (metabolism)
• Tumor Suppressor Protein p53 (deficiency, genetics)