Kinetic analyses of DNA‐linked ribonucleases H with different sizes of DNA

FEBS Letters

Uchiyama, Yohtaro¹  Kanaya, Shigenori²  Ikehara, Morio²  Ueno, Yoshio¹  Ohtsuka, Eiko³  Kanaya, Eiko² 

[1] Research Institute for Biosciences, Science University of Tokyo, 2669, Yamazaki, Noda, Chiba 278, Japan;Protein Engineering Research Institute, 6-2-3, Furuedai, Suita, Osaka 565, Japan;Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo 060, Japan

关键词: RNase H;    DNA-linked enzyme;    Kinetic study;    Site specificity;    RNase H;    ribonuclease H;    dX-mer;    Xbase DNA oligomer;    rX-mer;    X-base RNA oligomer;   

DOI  :  10.1016/0014-5793(94)01131-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown