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Rmi1 stimulates decatenation of double Holliday junctions during dissolution by Sgs1–Top3

Nature Structural & Molecular Biology volume 17, pages 1377–1382 (2010)Cite this article

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Abstract

A double Holliday junction (dHJ) is a central intermediate of homologous recombination that can be processed to yield crossover or non-crossover recombination products. To preserve genomic integrity, cells possess mechanisms to avoid crossing over. We show that Saccharomyces cerevisiae Sgs1 and Top3 proteins are sufficient to migrate and disentangle a dHJ to produce exclusively non-crossover recombination products, in a reaction termed “dissolution.” We show that Rmi1 stimulates dHJ dissolution at low Sgs1–Top3 protein concentrations, although it has no effect on the initial rate of Holliday junction (HJ) migration. Rmi1 serves to stimulate DNA decatenation, removing the last linkages between the repaired and template DNA molecules. Dissolution of a dHJ is a highly efficient and concerted alternative to nucleolytic resolution that prevents crossing over of chromosomes during recombinational DNA repair in mitotic cells and thereby contributes to genomic integrity.

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Figure 1: Sgs1 and Top3, stimulated by RPA, dissolve double Holliday junctions (dHJ) to yield non-crossover products.
Figure 2: Rmi1 stimulates a late step of dHJ dissolution catalyzed by Sgs1 and Top3.
Figure 3: Rmi1 greatly stimulates dissolution of DNA that contains dHJs separated by two topological links and that mimics a nearly dissolved dHJ.
Figure 4: Rmi1 promotes decatenation of kinetoplast DNA (kDNA) by Sgs1 and Top3.
Figure 5: Model for function of Sgs1–Top3–Rmi1 complex in the dissolution of double Holliday junctions (dHJs).

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Acknowledgements

We thank P. Janscak (University of Zurich), X. Veaute (Institute of Cellular and Molecular Radiation Biology, France), Tao-shih Hsieh (Duke University), B. Rad and A. Nimonkar (both University of California, Davis) for purified proteins and the members of the Kowalczykowski laboratory and W.D. Heyer (University of California, Davis) for their comments on the manuscript. This work was supported by the following grants: Swiss National Science Foundation Fellowship PA00A-115375 (P.C.), National Cancer Institute Award T32CA108459 (J.L.P.), Cancer Research UK (C.Z.B. and I.D.H.) and US National Institutes of Health grants GM-41347 and GM-62653 (S.C.K.).

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Author notes

  1. Petr Cejka and Jody L Plank: Both authors contributed equally to this study.

Authors and Affiliations

  1. Departments of Microbiology and Molecular and Cellular Biology, University of California, Davis, Davis, California, USA

    Petr Cejka, Jody L Plank & Stephen C Kowalczykowski

  2. Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, UK

    Csanad Z Bachrati & Ian D Hickson

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  1. Petr Cejka
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Contributions

P.C., J.L.P. and S.C.K. conceived the general ideas for this study. All authors planned experiments and interpreted data; J.L.P. prepared DHJS; P.C. and C.Z.B. performed experiments. P.C., J.L.P. and S.C.K. wrote the manuscript; and all authors provided editorial input.

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Correspondence to Stephen C Kowalczykowski.

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Cejka, P., Plank, J., Bachrati, C. et al. Rmi1 stimulates decatenation of double Holliday junctions during dissolution by Sgs1–Top3. Nat Struct Mol Biol 17, 1377–1382 (2010). https://doi.org/10.1038/nsmb.1919

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