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Title
Japanese: 
English:Analysis of substrate specificity of the RuvC Holliday junction resolvase with synthetic Holliday junctions 
Author
Japanese: Shida, T., 岩崎博史, Saito, A., Kyogoku, Y., Shinagawa, H..  
English: Shida, T., Hiroshi Iwasaki, Saito, A., Kyogoku, Y., Shinagawa, H..  
Language English 
Journal/Book name
Japanese:Journal of Biological Chemistry 
English:Journal of Biological Chemistry 
Volume, Number, Page Vol. 271    No. 42    pp. 26105-26109
Published date Oct. 1996 
Publisher
Japanese: 
English: 
Conference name
Japanese: 
English: 
Conference site
Japanese: 
English: 
Official URL http://www.scopus.com/inward/record.url?eid=2-s2.0-0029908324&partnerID=MN8TOARS
 
DOI https://doi.org/10.1074/jbc.271.42.26105
Abstract The Escherichia coli RuvC protein endonucleolytically resolves Holliday junctions, which are formed as intermediates during genetic recombination and recombination repair. Previous studies using model Holliday junctions suggested that a certain size of central core of homology and a specific sequence in the junction were required for efficient cleavage by RuvC, although not for binding. To determine the minimum length of sequence homology required for RuvC cleavage, we made a series of synthetic Holliday junctions with various lengths of homologous sequence in the core region. It was demonstrated that a monomobile junction possessing only 2 base pairs of the homology core was efficiently cleaved by RuvC. To study the sequence specificity for cleavage, we made 16 bimobile junctions, which differed only in the homologous core sequence. Among them, 6 junctions were efficiently cleaved. Cleavage occurred by introduction of nicks symmetrically at the 3'-side of thymine in all cases. However, the nucleotide bases at the 3'-side of the thymines were not always identical between the two strands nicked. These results suggest that RuvC recognizes mainly topological symmetry of the Holliday junction but not the sequence symmetry per se, that the thymine residue at the cleavage site plays an important role for RuvC-mediated resolution, and that a long homologous core sequence is not essential for cleavage.

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