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A Model of EcoRII Restriction Endonuclease Action: The Active Complex is Most Likely Formed by One Protein Subunit and One DNA Recognition Site
Author(s) -
Karpova Elizaveta A.,
Kubareva Elena A.,
Shabarova Zoe A.
Publication year - 1999
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/713803460
Subject(s) - endonuclease , restriction enzyme , dna , recognition sequence , protein subunit , biochemistry , biology , protein–dna interaction , chemistry , microbiology and biotechnology , dna binding protein , transcription factor , gene
To elucidate the mechanism of interaction of restriction endonuclease Eco RII with DNA, we studied by native gel electrophoresis the binding of this endonuclease to a set of synthetic DNA‐duplexes containing the modified orcanonical recognition sequence 5‐d(CCA/TGG)‐3. All binding substrate or substrate analogues tested could be divided into two major groups: (i) duplexes that, at the interaction with endonuclease EcoRII, form two types of stable complexes on native gel in the absence of Mg+2 cofactor; (ii) duplexes that form only one type of complex, observed both in the presence and absence of Mg+2. Unlike the latter, duplexes under the first group can be hydrolyzed by endonuclease. Data obtained suggest that the active complex is most likely formed by one protein subunit and one DNA recognition sequence. A model of EcoRII endonuclease action is presented.