A CRISPR-Cas9 screen identifies EXO1 as a formaldehyde resistance gene
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Dokumenter
- Fulltext
Forlagets udgivne version, 5,46 MB, PDF-dokument
Fanconi Anemia (FA) is a rare, genome instability-associated disease characterized by a deficiency in repairing DNA crosslinks, which are known to perturb several cellular processes, including DNA transcription, replication, and repair. Formaldehyde, a by-product of metabolism, is thought to drive FA by generating DNA interstrand crosslinks (ICLs) and DNA-protein crosslinks (DPCs). However, the impact of formaldehyde on global cellular pathways has not been investigated thoroughly. Herein, using a pangenomic CRISPR-Cas9 screen, we identify EXO1 as a critical regulator of formaldehyde-induced DNA lesions. We show that EXO1 knockout cell lines exhibit formaldehyde sensitivity leading to the accumulation of replicative stress, DNA double-strand breaks, and quadriradial chromosomes, a typical feature of FA. After formaldehyde exposure, EXO1 is recruited to chromatin, protects DNA replication forks from degradation, and functions in parallel with the FA pathway to promote cell survival. In vitro, EXO1-mediated exonuclease activity is proficient in removing DPCs. Collectively, we show that EXO1 limits replication stress and DNA damage to counteract formaldehyde-induced genome instability.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 381 |
| Tidsskrift | Nature Communications |
| Vol/bind | 14 |
| Udgave nummer | 1 |
| Antal sider | 20 |
| ISSN | 2041-1723 |
| DOI | |
| Status | Udgivet - 2023 |
Bibliografisk note
Funding Information:
The authors thank the Masson lab for critically reviewing the manuscript and for helpful comments. We thank Agata Smogorzewska, Detlev Shindler, Annamaria Ruggiano and Kristijan Ramadan, as well as Fatemeh Mashayekhi and Ismail Ismail for technical advice, Winfried Edelmann for Exo1 deficient MEFs, Josephine Dorsman for VU1365 and RPE2 cells, Alan d’Andrea for cell lines, Tom Moss for the use of a Leica SP5 microscope, and Yannick Doyon for the AAVS1 complementation system. L.G.S. was a FRQS postdoctoral fellow, and Y.G. was supported by Fondation du CHU de Québec and FRQS Ph.D. scholarships. This work was funded by grants from the European Research Council (grant no. 715975-DPC_REPAIR) to J.P.D and N.B.L., CIHR MOP-152948 to A.F.T., NSERC grant number RGPIN-2016-05847 to S.H. and CIHR FDN-388879 to J.Y.M. S.H. is an FRQS Junior 2 scholar, and A.F.T. is a Canada Research Chair in Molecular Virology and Genomic Instability and is supported by the Foundation J.-Louis Lévesque. J.Y.M. is a Tier I Canada Research Chair in DNA repair and Cancer Therapeutics.
Funding Information:
The authors thank the Masson lab for critically reviewing the manuscript and for helpful comments. We thank Agata Smogorzewska, Detlev Shindler, Annamaria Ruggiano and Kristijan Ramadan, as well as Fatemeh Mashayekhi and Ismail Ismail for technical advice, Winfried Edelmann for Exo1 deficient MEFs, Josephine Dorsman for VU1365 and RPE2 cells, Alan d’Andrea for cell lines, Tom Moss for the use of a Leica SP5 microscope, and Yannick Doyon for the AAVS1 complementation system. L.G.S. was a FRQS postdoctoral fellow, and Y.G. was supported by Fondation du CHU de Québec and FRQS Ph.D. scholarships. This work was funded by grants from the European Research Council (grant no. 715975-DPC_REPAIR) to J.P.D and N.B.L., CIHR MOP-152948 to A.F.T., NSERC grant number RGPIN-2016-05847 to S.H. and CIHR FDN-388879 to J.Y.M. S.H. is an FRQS Junior 2 scholar, and A.F.T. is a Canada Research Chair in Molecular Virology and Genomic Instability and is supported by the Foundation J.-Louis Lévesque. J.Y.M. is a Tier I Canada Research Chair in DNA repair and Cancer Therapeutics.
Publisher Copyright:
© 2023, The Author(s).
ID: 335965145