Kinetic analyses of DNA-linked ribonucleases H with different sizes of DNA.
A series of DNA-linked ribonucleases H with DNA adducts varying in size and sequence, ranging from heptamer to nonamer, were constructed and examined for their ability to cleave the 12-base RNA (5'-CGGAGAUGACGG-3') site-specifically. The DNA-linked RNase H with the 9-base DNA (5'-GTCATCTCC-3') cleaved the 12-base RNA specifically at A6-U7. Kinetic studies revealed that the DNA-linked RNase H with the 8-base DNA (5'-TCATCTCC-3') cleaved it slightly more effectively than that with the 9-base DNA. Factors that may affect the specificity and catalytic efficiency of a DNA-linked RNase H are described.
Kanaya
E
E
Protein Engineering Research Institute, Osaka, Japan.
Uchiyama
Y
Y
Ohtsuka
E
E
Ueno
Y
Y
Ikehara
M
M
Kanaya
S
S
D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism*; D009843:Oligoribonucleotides / Q000737:chemistry / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D016914:Ribonuclease H / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013696:Temperature
D009838:Oligodeoxyribonucleotides; D009843:Oligoribonucleotides; D012313:RNA; D004247:DNA; D016914:Ribonuclease H