Abstract

DNA end resection plays a critical function in DNA double-strand break repair pathway choice. Resected DNA ends are refractory to end-joining mechanisms and are instead channeled to homology-directed repair. Using biochemical, genetic, and imaging methods, we show that phosphorylation of Saccharomyces cerevisiae Sae2 controls its capacity to promote the Mre11-Rad50-Xrs2 (MRX) nuclease to initiate resection of blocked DNA ends by at least two distinct mechanisms. First, DNA damage and cell cycle-dependent phosphorylation leads to Sae2 tetramerization. Second, and independently, phosphorylation of the conserved C-terminal domain of Sae2 is a prerequisite for its physical interaction with Rad50, which is also crucial to promote the MRX endonuclease. The lack of this interaction explains the phenotype of rad50S mutants defective in the processing of Spo11-bound DNA ends during meiotic recombination. Our results define how phosphorylation controls the initiation of DNA end resection and therefore the choice between the key DNA double-strand break repair mechanisms.

Details

Title
Regulatory control of DNA end resection by Sae2 phosphorylation
Author
Cannavo, Elda 1 ; Johnson, Dominic 2 ; Andres, Sara N 3 ; Kissling, Vera M 4 ; Reinert, Julia K 5   VIAFID ORCID Logo  ; Garcia, Valerie 6 ; Erie, Dorothy A 7 ; Hess, Daniel 8   VIAFID ORCID Logo  ; Thomä, Nicolas H 8   VIAFID ORCID Logo  ; Enchev, Radoslav I 4 ; Matthias, Peter 4   VIAFID ORCID Logo  ; Williams, R Scott 9 ; Neale, Matt J 2 ; Cejka, Petr 10 

1  Faculty of Biomedical Sciences, Institute for Research in Biomedicine, Università della Svizzera italiana (USI), Bellinzona, Switzerland 
2  Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton, UK 
3  Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, Department of Health and Human Services, US National Institutes of Health, Research Triangle Park, NC, USA; Michael G. DeGroote Institute for Infectious Disease Research, Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada 
4  Department of Biology, Institute of Biochemistry, Eidgenössische Technische Hochschule (ETH), Zürich, Switzerland 
5  Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland; University of Basel, Basel, Switzerland 
6  Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton, UK; Centre de Recherche en Cancérologie de Marseille (CRCM), Institut Paoli Calmettes, Inserm UMR1068, CNRS UMR7258, Aix Marseille Université, Marseille, France 
7  Department of Chemistry, Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC, USA 
8  Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland 
9  Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, Department of Health and Human Services, US National Institutes of Health, Research Triangle Park, NC, USA 
10  Faculty of Biomedical Sciences, Institute for Research in Biomedicine, Università della Svizzera italiana (USI), Bellinzona, Switzerland; Department of Biology, Institute of Biochemistry, Eidgenössische Technische Hochschule (ETH), Zürich, Switzerland 
Pages
1-14
Publication year
2018
Publication date
Oct 2018
Publisher
Nature Publishing Group
e-ISSN
20411723
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41467-018-06417-5
ProQuest document ID
2115238542
Copyright
© 2018. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.