Cleavage of deoxyoxanosine-containing oligodeoxyribonucleotides by bacterial endonuclease V.
Oxanine (O) is a deamination product derived from guanine with the nitrogen at the N1 position substituted by oxygen. Cytosine, thymine, adenine, guanine as well as oxanine itself can be incorporated by Klenow Fragment to pair with oxanine in a DNA template with similar efficiency, indicating that oxanine in DNA may cause various mutations. As a nucleotide, deoxyoxanosine may substitute for deoxyguanosine to complete a primer extension reaction. Endonuclease V, an enzyme known for its enzymatic activity on uridine-, inosine- and xanthosine-containing DNA, can cleave oxanosine-containing DNA at the second phosphodiester bond 3' to the lesion. Mg2+ or Mn2+, and to a small extent Co2+ or Ni2+, support the oxanosine-containing DNA cleavage activity. All four oxanosine-containing base pairs (A/O, T/O, C/O and G/O) were cleaved with similar efficiency. The cleavage of double-stranded oxanosine-containing DNA was approximately 6-fold less efficient than that of double-stranded inosine-containing DNA. Single-stranded oxanosine-containing DNA was cleaved with a lower efficiency as compared with double-stranded oxanosine-containing DNA. A metal ion enhances the binding of endonuclease V to double-stranded and single-stranded oxanosine-containing DNA 6- and 4-fold, respectively. Hypothetic models of oxanine-containing base pairs and deaminated base recognition mechanism are presented.
Department of Genetics, Biochemistry and Life Science Studies, South Carolina, Experiment Station, Clemson University, Room 219, Biosystems Research Complex, 51 New Cherry Street, Clemson, SC 29634, USA.
Thomas M
TM
Hitchcock
Honghai
H
Gao
Weiguo
W
Cao
D001419:Bacteria / Q000201:enzymology*; D020029:Base Pairing; D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism; D043224:Deoxyribonuclease (Pyrimidine Dimer) / Q000378:metabolism*; D003853:Deoxyribonucleosides / Q000737:chemistry*; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D008956:Models, Chemical; D016296:Mutagenesis; D009838:Oligodeoxyribonucleotides / Q000737:chemistry* / Q000378:metabolism*; D011684:Purine Nucleosides / Q000737:chemistry; D013379:Substrate Specificity
D004277:DNA, Single-Stranded; D003853:Deoxyribonucleosides; D008670:Metals; D009838:Oligodeoxyribonucleotides; D011684:Purine Nucleosides; C468766:oxanine; C042917:2'-deoxyoxanosine; D043224:Deoxyribonuclease (Pyrimidine Dimer)