Yeast Mph1 helicase dissociates Rad51-made D-loops: implications for crossover control in mitotic recombination

  1. Rohit Prakash1,7,
  2. Dominik Satory2,7,
  3. Eloïse Dray1,
  4. Almas Papusha2,
  5. Jürgen Scheller3,8,
  6. Wilfried Kramer3,
  7. Lumir Krejci4,
  8. Hannah Klein5,
  9. James E. Haber6,
  10. Patrick Sung1,10 and
  11. Grzegorz Ira2,9
  1. 1Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06520, USA
  2. 2Baylor College of Medicine, Department of Molecular and Human Genetics One Baylor Plaza, Houston, Texas 77030, USA
  3. 3Institute for Microbiology and Genetics, University of Göttingen, Department of Molecular Genetics and Preparative Molecular Biology, D-37077 Göttingen, Germany
  4. 4National Center for Biomolecular Research, Masaryk University, Brno 62500, Czech Republic
  5. 5Department of Biochemistry and Kaplan Cancer Center, New York University School of Medicine, New York, New York 10016, USA
  6. 6Rosenstiel Center and Department of Biology, Brandeis University, Waltham, Massachusetts 02454, USA

    1. ↵ These authors contributed equally to this work

    Abstract

    Eukaryotes possess mechanisms to limit crossing over during homologous recombination, thus avoiding possible chromosomal rearrangements. We show here that budding yeast Mph1, an ortholog of human FancM helicase, utilizes its helicase activity to suppress spontaneous unequal sister chromatid exchanges and DNA double-strand break-induced chromosome crossovers. Since the efficiency and kinetics of break repair are unaffected, Mph1 appears to channel repair intermediates into a noncrossover pathway. Importantly, Mph1 works independently of two other helicases—Srs2 and Sgs1—that also attenuate crossing over. By chromatin immunoprecipitation, we find targeting of Mph1 to double-strand breaks in cells. Purified Mph1 binds D-loop structures and is particularly adept at unwinding these structures. Importantly, Mph1, but not a helicase-defective variant, dissociates Rad51-made D-loops. Overall, the results from our analyses suggest a new role of Mph1 in promoting the noncrossover repair of DNA double-strand breaks.

    Keywords:

    Keywords

    Footnotes

    • ↵ Present address: Institute of Biochemistry, Medical Faculty, Christian Albrecht, University Kiel, Olshausenstrasse 40, D-24098 Kiel, Germany

    • ↵ Corresponding authors.

      ↵ E-MAIL gira{at}bcm.edu; FAX (713) 798-8967.

    • ↵ E-MAIL patrick.sung{at}yale.edu.; FAX (203) 785 6037.

    • Article is online at http://www.genesdev.org/cgi/doi/10.1101/gad.1737809.

    • Supplemental material is available at http://www.genesdev.org.

      • Received September 8, 2008.
      • Accepted November 12, 2008.
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    1. doi: 10.1101/gad.1737809 Genes & Dev. 23: 67-79 Copyright © 2009 by Cold Spring Harbor Laboratory Press

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