Identification of Holliday junction resolvases from humans and yeast

Ip, Stephen C Y, Rass, Ulrich, Blanco, Miguel G, Flynn, Helen R, Skehel, J Mark and West, Stephen C (2008) Identification of Holliday junction resolvases from humans and yeast. Nature, 456 (7220). pp. 357-61. ISSN 0028-0836

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Four-way DNA intermediates, also known as Holliday junctions, are formed during homologous recombination and DNA repair, and their resolution is necessary for proper chromosome segregation. Here we identify nucleases from Saccharomyces cerevisiae and human cells that promote Holliday junction resolution, in a manner analogous to that shown by the Escherichia coli Holliday junction resolvase RuvC. The human Holliday junction resolvase, GEN1, and its yeast orthologue, Yen1, were independently identified using two distinct experimental approaches: GEN1 was identified by mass spectrometry following extensive fractionation of HeLa cell-free extracts, whereas Yen1 was detected by screening a yeast gene fusion library for nucleases capable of Holliday junction resolution. The eukaryotic Holliday junction resolvases represent a new subclass of the Rad2/XPG family of nucleases. Recombinant GEN1 and Yen1 resolve Holliday junctions by the introduction of symmetrically related cuts across the junction point, to produce nicked duplex products in which the nicks can be readily ligated.

Item Type: Article
Keywords: DNA repair, homologous recombination, DNA double-strand break repair, replication stress, chromosome break repair, Holliday junction resolution, structure-specific nucleases, Yen1, GEN1, Slx1, Slx4, Mus81, Mms4
Schools and Departments: School of Life Sciences > Sussex Centre for Genome Damage and Stability
Research Centres and Groups: Genome Damage and Stability Centre
Subjects: Q Science > Q Science (General) > Q0179.9 Research
Q Science > Q Science (General)
Depositing User: Ulrich Rass
Date Deposited: 26 Sep 2018 08:05
Last Modified: 26 Sep 2018 08:05
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