Restriction endonuclease BsoFI is sensitive to the 5'-methylation of deoxycytidines in its recognition sequence

Nucleic Acids Res. 1995 Nov 11;23(21):4227-8. doi: 10.1093/nar/23.21.4227.

Abstract

The isoschizomeric restriction endonucleases Fnu4HI and BsoFI cleave DNA at 5'-GCdecreasesNGC-3' sequences. Fnu4HI has been shown to be inhibited by 5'-CG-3'methylation in the sequences 5'-GmCGGC-3' or 5'-GCGGmCG-3'. We have now investigated the methylation sensitivity of BsoFI by testing its activity on plasmid DNA 5'-CG-3' methylated with the M.SssI DNA methyltransferase or on synthetic (CGG)n repetitive oligodeoxyribonucleotides which have been partly or completely C methylated. The data demonstrate that BsoFI cannot cleave at its recognition sequence when it is completely 5'-CG-3' methylated. These enzymes have proven to be useful in analyses of the methylation status in (CGG)n repeats of the human genome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA / metabolism*
  • Deoxycytidine / analogs & derivatives
  • Deoxyribonucleases, Type II Site-Specific / metabolism*
  • Methylation
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides / metabolism*
  • Substrate Specificity

Substances

  • Oligodeoxyribonucleotides
  • Deoxycytidine
  • DNA
  • 5-methyldeoxycytidine
  • endodeoxyribonuclease Fnu4HI
  • Deoxyribonucleases, Type II Site-Specific