Molecular insights into the activation of Mre11-Rad50 endonuclease activity by Sae2/CtIP
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Abstract
In Saccharomyces cerevisiae (S. cerevisiae), Mre11-Rad50-Xrs2 (MRX)-Sae2 nuclease activity is required for the resection of DNA breaks with secondary structures or protein blocks, while in humans, the MRE11-RAD50-NBS1 (MRN) homolog with CtIP is needed to initiate DNA end resection of all breaks. Phosphorylated Sae2/CtIP stimulates the endonuclease activity of MRX/N. Structural insights into the activation of the Mre11 nuclease are available only for organisms lacking Sae2/CtIP, so little is known about how Sae2/CtIP activates the nuclease ensemble. Here, we uncover the mechanism of Mre11 activation by Sae2 using a combination of AlphaFold2 structural modeling of biochemical and genetic assays. We show that Sae2 stabilizes the Mre11 nuclease in a conformation poised to cleave substrate DNA. Several designs of compensatory mutations establish how Sae2 activates MRX in vitro and in vivo, supporting the structural model. Finally, our study uncovers how human CtIP, despite considerable sequence divergence, employs a similar mechanism to activate MRN.
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Mattarocci, S., Reginato, G., Cannavò, E., Roisné-Hamelin, F., Gnügge, R., Deshayes, A., Pobiega, S., Alibert, O., Morin, V., Ropars, V., et al. (2026) · Nature Structural & Molecular Biology · doi:10.1038/s41594-026-01810-x · Cited by 1 · see citing works
Casari, E., Gnugnoli, M., Pizzul, P., Tisi, R., & Longhese, M. P. (2025) · Communications Biology · doi:10.1038/s42003-025-09424-7
Discovery of a FANCD2-interacting protein motif (DIP-box) linking DNA Damage Response processesPreprint
Cao, Z.-B., Briola, G., Ionita, C., Streetley, J., Walden, H., & Rennie, M. (2026) · bioRxiv · doi:10.64898/2026.02.24.707417
- Authors
- Nicolas, Y., Bret, H., Cannavo, E., Acharya, A., Cejka, P., Borde, V., & Guerois, R.
- Affiliations
- Centre National de la Recherche Scientifique (FR)
- Institute for Research in Biomedicine (CH)
- Corresponding author
- Borde, Valérie
- Venue
- Molecular Cell
- Publisher
- Elsevier BV
- Published
- 2024-06
- Volume
- 84
- Issue
- 12
- Pages
- 2223-2237.e4
- ISSN
- 1097-2765 (ISSN-L)
- OpenAlex
- W4399566917
- Cited by
- 12
- References
- 81
- Citations by year
- 2026: 2 · 2025: 7 · 2024: 3
- FWCI
- 1.64
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.elsevier.com/tdm/userlicense/1.0/ (tdm)
- www.elsevier.com/legal/tdmrep-license (tdm)
- www.elsevier.com/open-access/userlicense/1.0/ (vor, 384 d)
- doi.org/10.15223/policy-017 (stm-asf)
- doi.org/10.15223/policy-037 (stm-asf)
- doi.org/10.15223/policy-012 (stm-asf)
- doi.org/10.15223/policy-029 (stm-asf)
- doi.org/10.15223/policy-004 (stm-asf)
- Locations
- Molecular Cell (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nd)
- PubMed (publishedVersion)
- HAL (submittedVersion, other-oa)
- HAL (submittedVersion)
- Funders
- ERC (101018257)
- FRMD
- ANR (ANR-21-CE44-0009)
- Fondation pour la Recherche Médicale (FRM EQU201903007785)
- SNSF (310030_205199, 310030_207588)
- Commissariat à l'Énergie Atomique et aux Énergies Alternatives
- Grand Équipement National De Calcul Intensif
- European Research Council
- University of Sussex
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Keywords
- MRX complex, MRN complex, DNA end resection, nuclease activation, protein-DNA complex structure, compensatory mutations, Saccharomyces cerevisiae, DNA double-strand break repair
- MeSH terms
- MRE11 Homologue Protein, DNA Repair, DNA-Binding Proteins, Endodeoxyribonucleases, Endonucleases, Enzyme Activation, Exodeoxyribonucleases, Humans, Models, Molecular, Mutation, Phosphorylation, Saccharomyces cerevisiae, Acid Anhydride Hydrolases, Saccharomyces cerevisiae Proteins, DNA Repair Enzymes, DNA Breaks, Double-Stranded
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