Cleavage of a DNA-RNA-DNA/DNA chimeric substrate containing a single ribonucleotide at the DNA-RNA junction with prokaryotic RNases HII.
We have analyzed the cleavage specificities of various prokaryotic Type 2 ribonucleases H (RNases H) on chimeric DNA-RNA-DNA/DNA substrates containing one to four ribonucleotides. RNases HII from Bacillus subtilis and Thermococcus kodakaraensis cleaved all of these substrates to produce a DNA segment with a 5'-monoribonucleotide. Consequently, these enzymes cleaved even the chimeric substrate containing a single ribonucleotide at the DNA-RNA junction (5'-side of the single ribonucleotide). In contrast, Escherichia coli RNase HI and B. subtilis RNase HIII did not cleave the chimeric substrate containing a single ribonucleotide. These results suggest that bacterial and archaeal RNases HII are involved in excision of a single ribonucleotide misincorporated into DNA.
Haruki
Mitsuru
M
Department of Material and Life Science, Graduate School of Engineering, Osaka University, 2-1, Yamadaoka, Suita, Osaka, Japan.
Tsunaka
Yasuo
Y
Morikawa
Masaaki
M
Kanaya
Shigenori
S
D001412:Bacillus subtilis / Q000201:enzymology; D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D007700:Kinetics; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D016914:Ribonuclease H / Q000378:metabolism*; D012265:Ribonucleotides / Q000378:metabolism; D013379:Substrate Specificity; D019712:Thermococcus / Q000201:enzymology
D009692:Nucleic Acid Heteroduplexes; D012265:Ribonucleotides; D012313:RNA; D004247:DNA; C091139:ribonuclease HII; D016914:Ribonuclease H; C078856:ribonuclease HI