S-adenosyl homocysteine and DNA ends stimulate promiscuous nuclease activities in the Type III restriction endonuclease EcoPI

Nucleic Acids Res. 2009 Jul;37(12):3934-45. doi: 10.1093/nar/gkp267. Epub 2009 Apr 28.

Abstract

In the absence of the methyl donor S-adenosyl methionine and under certain permissive reaction conditions, EcoPI shows non-specific endonuclease activity. We show here that the cofactor analogue S-adenosyl homocysteine promotes this promiscuous DNA cleavage. Additionally, an extensive exonuclease-like processing of the DNA is also observed that can even result in digestion of non-specific DNA in trans. We suggest a model for how DNA communication events initiating from non-specific sites, and in particular free DNA ends, could produce the observed cleavage patterns.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Catalytic Domain
  • DNA / chemistry
  • DNA / metabolism*
  • DNA Cleavage*
  • Deoxyribonucleases, Type III Site-Specific / chemistry
  • Deoxyribonucleases, Type III Site-Specific / metabolism*
  • Exodeoxyribonucleases / metabolism*
  • S-Adenosylhomocysteine / metabolism*

Substances

  • Adenosine Triphosphate
  • DNA
  • S-Adenosylhomocysteine
  • Exodeoxyribonucleases
  • DNA restriction enzyme EcoPI
  • Deoxyribonucleases, Type III Site-Specific