Dna2 nuclease-helicase structure, mechanism and regulation by Rpa

  1. Chun Zhou
  2. Sergei Pourmal
  3. Nikola P Pavletich  Is a corresponding author
  1. Memorial Sloan Kettering Cancer Center, United States

Abstract

The Dna2 nuclease-helicase maintains genomic integrity by processing DNA double-strand breaks, Okazaki fragments and stalled replication forks. Dna2 requires ssDNA ends, and is dependent on the ssDNA-binding protein Rpa, which controls cleavage polarity. Here we present the 2.3 Å structure of intact mouse Dna2 bound to a 15-nucleotide ssDNA. The nuclease active site is embedded in a long, narrow tunnel through which the DNA has to thread. The helicase domain is required for DNA binding but not threading. We also present the structure of a flexibly-tethered Dna2-Rpa interaction that recruits Dna2 to Rpa-coated DNA. We establish that a second Dna2-Rpa interaction is mutually exclusive with Rpa-DNA interactions and mediates the displacement of Rpa from ssDNA. This interaction occurs at the nuclease tunnel entrance and the 5' end of the Rpa-DNA complex. Hence, it only displaces Rpa from the 5' but not 3' end, explaining how Rpa regulates cleavage polarity.

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Author details

  1. Chun Zhou

    Structural Biology Program, Memorial Sloan Kettering Cancer Center, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Sergei Pourmal

    Structural Biology Program, Memorial Sloan Kettering Cancer Center, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Nikola P Pavletich

    Structural Biology Program, Memorial Sloan Kettering Cancer Center, New York, United States
    For correspondence
    pavletin@mskcc.org
    Competing interests
    The authors declare that no competing interests exist.

Copyright

© 2015, Zhou 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. Chun Zhou
  2. Sergei Pourmal
  3. Nikola P Pavletich
(2015)
Dna2 nuclease-helicase structure, mechanism and regulation by Rpa
eLife 4:e09832.
https://doi.org/10.7554/eLife.09832

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https://doi.org/10.7554/eLife.09832