2. Academic Validation
  3. FBH1 helicase disrupts RAD51 filaments in vitro and modulates homologous recombination in mammalian cells

FBH1 helicase disrupts RAD51 filaments in vitro and modulates homologous recombination in mammalian cells

  • J Biol Chem. 2013 Nov 22;288(47):34168-34180. doi: 10.1074/jbc.M113.484493.
Jitka Simandlova 1,  Jennifer Zagelbaum 2,  Miranda J Payne 3,  Wai Kit Chu 4,  Igor Shevelev 1,  Katsuhiro Hanada 3,  Sujoy Chatterjee 2,  Dylan A Reid 2,  Ying Liu 5,  Pavel Janscak 6,  Eli Rothenberg 7,  Ian D Hickson 8
Affiliations

Affiliations

  • 1 Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, 14300 Prague, Czech Republic.
  • 2 Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, New York.
  • 3 Weatherall Institute of Molecular Medicine, Department of Oncology, University of Oxford, Oxford OX3 9DS, United Kingdom.
  • 4 Weatherall Institute of Molecular Medicine, Department of Oncology, University of Oxford, Oxford OX3 9DS, United Kingdom; Nordea Center for Healthy Aging, Department of Cellular and Molecular Medicine, University of Copenhagen, 2200 Copenhagen N, Denmark.
  • 5 Nordea Center for Healthy Aging, Department of Cellular and Molecular Medicine, University of Copenhagen, 2200 Copenhagen N, Denmark.
  • 6 Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, 14300 Prague, Czech Republic; Institute of Molecular Cancer Research, University of Zurich, CH-8057 Zurich, Switzerland. Electronic address: pjanscak@imcr.uzh.ch.
  • 7 Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, New York. Electronic address: eli.rothenberg@nyumc.org.
  • 8 Weatherall Institute of Molecular Medicine, Department of Oncology, University of Oxford, Oxford OX3 9DS, United Kingdom; Nordea Center for Healthy Aging, Department of Cellular and Molecular Medicine, University of Copenhagen, 2200 Copenhagen N, Denmark. Electronic address: iandh@sund.ku.dk.
PMID: 24108124 DOI: 10.1074/jbc.M113.484493
Abstract

Efficient repair of DNA double strand breaks and interstrand cross-links requires the homologous recombination (HR) pathway, a potentially error-free process that utilizes a homologous sequence as a repair template. A key player in HR is RAD51, the eukaryotic ortholog of Bacterial RecA protein. RAD51 can polymerize on DNA to form a nucleoprotein filament that facilitates both the search for the homologous DNA sequences and the subsequent DNA strand invasion required to initiate HR. Because of its pivotal role in HR, RAD51 is subject to numerous positive and negative regulatory influences. Using a combination of molecular genetic, biochemical, and single-molecule biophysical techniques, we provide mechanistic insight into the mode of action of the FBH1 helicase as a regulator of RAD51-dependent HR in mammalian cells. We show that FBH1 binds directly to RAD51 and is able to disrupt RAD51 filaments on DNA through its ssDNA translocase function. Consistent with this, a mutant mouse embryonic stem cell line with a deletion in the FBH1 helicase domain fails to limit RAD51 chromatin association and shows hyper-recombination. Our data are consistent with FBH1 restraining RAD51 DNA binding under unperturbed growth conditions to prevent unwanted or unscheduled DNA recombination.

Keywords

DNA Damage; DNA Helicase; DNA Recombination; DNA Repair; DNA Replication.

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