MutS/MutL crystal structure reveals that the MutS sliding clamp loads MutL onto DNA

  1. Flora S Groothuizen
  2. Ines Winkler
  3. Michele Cristóvão
  4. Alexander Fish
  5. Herrie H K Winterwerp
  6. Annet Reumer
  7. Andreas D Marx
  8. Nicolaas Hermans
  9. Robert A Nicholls
  10. Garib N Murshudov
  11. Joyce H G Lebbink
  12. Peter Friedhoff
  13. Titia K Sixma  Is a corresponding author
  1. Netherlands Cancer Institute, Netherlands
  2. Justus-Liebig-University, Germany
  3. Erasmus Medical Center, Netherlands
  4. MRC Laboratory of Molecular Biology, United Kingdom

Abstract

To avoid mutations in the genome, DNA replication is generally followed by DNA mismatch repair (MMR). MMR starts when a MutS homolog recognizes a mismatch and undergoes an ATP-dependent transformation to an elusive sliding clamp state. How this transient state promotes MutL homolog recruitment and activation of repair is unclear. Here we present a crystal structure of the MutS/MutL complex using a site-specifically crosslinked complex and examine how large conformational changes lead to activation of MutL. The structure captures MutS in the sliding clamp conformation, where tilting of the MutS subunits across each other pushes DNA into a new channel, and reorientation of the connector domain creates an interface for MutL with both MutS subunits. Our work explains how the sliding clamp promotes loading of MutL onto DNA, to activate downstream effectors. We thus elucidate a crucial mechanism that ensures that MMR is initiated only after detection of a DNA mismatch.

Article and author information

Author details

  1. Flora S Groothuizen

    Division of Biochemistry and CGC.nl, Netherlands Cancer Institute, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  2. Ines Winkler

    Institute for Biochemistry, Justus-Liebig-University, Giessen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Michele Cristóvão

    Institute for Biochemistry, Justus-Liebig-University, Giessen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Alexander Fish

    Division of Biochemistry and CGC.nl, Netherlands Cancer Institute, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  5. Herrie H K Winterwerp

    Division of Biochemistry and CGC.nl, Netherlands Cancer Institute, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  6. Annet Reumer

    Division of Biochemistry and CGC.nl, Netherlands Cancer Institute, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  7. Andreas D Marx

    Institute for Biochemistry, Justus-Liebig-University, Giessen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Nicolaas Hermans

    Department of Genetics, Cancer Genomics Netherlands, Erasmus Medical Center, Rotterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  9. Robert A Nicholls

    Structural Studies Division, MRC Laboratory of Molecular Biology, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  10. Garib N Murshudov

    Structural Studies Division, MRC Laboratory of Molecular Biology, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  11. Joyce H G Lebbink

    Department of Genetics, Cancer Genomics Netherlands, Erasmus Medical Center, Rotterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  12. Peter Friedhoff

    Institute for Biochemistry, Justus-Liebig-University, Giessen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  13. Titia K Sixma

    Division of Biochemistry and CGC.nl, Netherlands Cancer Institute, Amsterdam, Netherlands
    For correspondence
    t.sixma@nki.nl
    Competing interests
    The authors declare that no competing interests exist.

Copyright

© 2015, Groothuizen et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Flora S Groothuizen
  2. Ines Winkler
  3. Michele Cristóvão
  4. Alexander Fish
  5. Herrie H K Winterwerp
  6. Annet Reumer
  7. Andreas D Marx
  8. Nicolaas Hermans
  9. Robert A Nicholls
  10. Garib N Murshudov
  11. Joyce H G Lebbink
  12. Peter Friedhoff
  13. Titia K Sixma
(2015)
MutS/MutL crystal structure reveals that the MutS sliding clamp loads MutL onto DNA
eLife 4:e06744.
https://doi.org/10.7554/eLife.06744

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https://doi.org/10.7554/eLife.06744

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