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Kinetic analyses of DNA‐linked ribonucleases H with different sizes of DNA
Author(s) -
Kanaya Eiko,
Uchiyama Yohtaro,
Ohtsuka Eiko,
Ueno Yoshio,
Ikehara Morio,
Kanaya Shigenori
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)01131-1
Subject(s) - dna , rnase p , rnase h , base pair , cleave , rna , ribonuclease , microbiology and biotechnology , biology , chemistry , biochemistry , gene
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.