Purification and characterization of a novel UV lesion-specific DNA glycosylase/AP lyase from Bacillus sphaericus

Debra A. Vasquez, Simon G. Nyaga, R. Stephen Lloyd

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


The purification and characterization of a pyrimidine dimer-specific glycosylase/AP lyase from Bacillus sphaericus (Bsp-pdg) are reported. Bsp-pdg is highly specific for DNA containing the cis-syn cyclobutane pyrimidine dimer, displaying no detectable activity on oligonucleotides with trans-syn I, trans-syn II, (6-4), or Dewar photoproducts. Like other glycosylase/AP lyases that sequentially cleave the N-glycosyl bond of the 5' pyrimidine of a cyclobutane pyrimidine dimer, and the phosphodiester backbone, this enzyme appears to utilize a primary amine as the attacking nucleophile. The formation of a covalent enzyme-DNA imino intermediate is evidenced by the ability to trap this protein-DNA complex by reduction with sodium borohydride. Also consistent with its AP lyase activity, Bsp-pdg was shown to incise an AP site-containing oligonucleotide, yielding β- and δ-elimination products. N-terminal amino acid sequence analysis of this 26 kDa protein revealed little amino acid homology to any previously reported protein. This is the first report of a glycosylase/AP lyase enzyme from Bacillus sphaericus that is specific for cis-syn pyrimidine dimers. Copyright (C) 2000 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)307-316
Number of pages10
JournalMutation Research - DNA Repair
Issue number4
StatePublished - May 31 2000
Externally publishedYes


  • Bacillus sphaericus
  • Glycosylase/AP lyase
  • Pyrimidine dimer

ASJC Scopus subject areas

  • Molecular Biology
  • Toxicology
  • Genetics


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