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Separase-mediated cleavage of cohesin at interphase is required for DNA repair

Abstract

Sister chromatids are held together by cohesins1. At anaphase, separase is activated by degradation of its inhibitory partner, securin2,3. Separase then cleaves cohesins4,5,6, thus allowing sister chromatid separation. Fission yeast securin (Cut2) has destruction boxes and a separase (Cut1) interaction site in the amino and carboxyl terminus, respectively7,8. Here we show that securin is essential for separase stability and also for proper repair of DNA damaged by ultraviolet, X-ray and γ-ray irradiation. The cut2EA2 mutant is defective in the repair of ultraviolet damage lesions, although the DNA damage checkpoint is activated normally. In double mutant analysis of ultraviolet sensitivity, checkpoint kinase chk1 (ref. 9) and excision repair rad13 (ref. 10) mutants were additive with cut2EA2, whereas recombination repair rhp51 (ref. 11) and cohesin subunit rad21 (ref. 12) mutants were not. Cohesin was hyper-modified on ultraviolet irradiation in a Rad3 kinase-dependent way13. Experiments using either mutant cohesin that cannot be cleaved by separase or a protease-dead separase provide evidence that this DNA repair function of securin–separase acts through the cleavage of cohesin. We propose that the securin–separase complex might aid DNA repair by removing local cohesin in interphase cells.

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Figure 1: Securin mutant cut2EA2 is defective in DNA damage repair in checkpoint-arrested cells.
Figure 2: Ultraviolet sensitivity of cut2EA2 is additive with Δchk1, but not with Δrhp51.
Figure 3: The DNA damage repair role of securin–separase acts through Rad21 cohesin.
Figure 4: Rad21 cohesin is hyper-modified in response to ultraviolet irradiation.

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Acknowledgements

We would like to thank C. Shimoda and T. Nakamura for the S. pombe genomic library, T. Carr, H. Okayama and N. Walworth for strains, and T. Matsumoto for introducing X-ray and γ-ray irradiation. This study was supported by the COE Grant of the Ministry of Education, Culture, Sports, Science and Technology of Japan. K.N. was a recipient of a predoctoral fellowship from the Japan Society for Promotion of Science.

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Correspondence to Mitsuhiro Yanagida.

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Nagao, K., Adachi, Y. & Yanagida, M. Separase-mediated cleavage of cohesin at interphase is required for DNA repair. Nature 430, 1044–1048 (2004). https://doi.org/10.1038/nature02803

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