The helicase-like domains of type III restriction enzymes trigger long-range diffusion along DNA

Science. 2013 Apr 19;340(6130):353-6. doi: 10.1126/science.1231122.

Abstract

Helicases are ubiquitous adenosine triphosphatases (ATPases) with widespread roles in genome metabolism. Here, we report a previously undescribed functionality for ATPases with helicase-like domains; namely, that ATP hydrolysis can trigger ATP-independent long-range protein diffusion on DNA in one dimension (1D). Specifically, using single-molecule fluorescence microscopy we show that the Type III restriction enzyme EcoP15I uses its ATPase to switch into a distinct structural state that diffuses on DNA over long distances and long times. The switching occurs only upon binding to the target site and requires hydrolysis of ~30 ATPs. We define the mechanism for these enzymes and show how ATPase activity is involved in DNA target site verification and 1D signaling, roles that are common in DNA metabolism: for example, in nucleotide excision and mismatch repair.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism*
  • DNA / chemistry
  • DNA / metabolism*
  • DNA Cleavage*
  • DNA Helicases / chemistry
  • DNA Helicases / metabolism*
  • Deoxyribonucleases, Type III Site-Specific / chemistry
  • Deoxyribonucleases, Type III Site-Specific / metabolism*
  • Hydrolysis
  • Microscopy, Fluorescence / methods
  • Nucleic Acid Conformation
  • Protein Structure, Tertiary

Substances

  • Adenosine Triphosphate
  • DNA
  • endodeoxyribonuclease EcoP15I
  • Deoxyribonucleases, Type III Site-Specific
  • DNA Helicases