Polymerase synthesis of DNAs bearing vinyl groups in the major groove and their cleavage by restriction endonucleases

Chembiochem. 2014 Oct 13;15(15):2306-12. doi: 10.1002/cbic.201402319. Epub 2014 Sep 1.

Abstract

DNA molecules containing 5-vinyluracil, 5-vinylcytosine, or 7-deaza-7-vinyladenine were prepared by polymerase incorporation of the corresponding vinyl-modified 2'-deoxyribonucleoside triphosphates, and the influence of the vinyl group in the major groove of DNA on the cleavage by diverse type II restriction endonucleases (REs) was studied. The presence of 5-vinyluracil was tolerated by most of the REs, whereas only some REs were able to cleave sequences containing 7-deaza-7-vinyladenine. The enzyme ScaI was found to cleave DNA containing 5-vinylcytosine efficiently but not DNA containing the related 5-ethynylcytosine. All other REs failed to cleave sequences containing any cytosine modifications.

Keywords: DNA; DNA polymerases; DNA-protein interactions; nucleotides; restriction endonucleases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / biosynthesis*
  • DNA / chemistry*
  • DNA / metabolism
  • DNA Cleavage
  • DNA Restriction Enzymes / metabolism*

Substances

  • DNA
  • DNA Restriction Enzymes