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Escherichia coli RuvC protein is an endonuclease that resolves the Holliday structure.

1991; Springer Nature; Volume: 10; Issue: 13 Linguagem: Inglês

10.1002/j.1460-2075.1991.tb05016.x

1460-2075

Hiroshi Iwasaki, Masahiko Takahagi, Toshikazu Shiba, Akio Nakata, Hideo Shinagawa,

RNA and protein synthesis mechanisms

Research Article1 December 1991free access Escherichia coli RuvC protein is an endonuclease that resolves the Holliday structure. H. Iwasaki H. Iwasaki Department of Experimental Chemotherapy, Osaka University, Japan. Search for more papers by this author M. Takahagi M. Takahagi Department of Experimental Chemotherapy, Osaka University, Japan. Search for more papers by this author T. Shiba T. Shiba Department of Experimental Chemotherapy, Osaka University, Japan. Search for more papers by this author A. Nakata A. Nakata Department of Experimental Chemotherapy, Osaka University, Japan. Search for more papers by this author H. Shinagawa H. Shinagawa Department of Experimental Chemotherapy, Osaka University, Japan. Search for more papers by this author H. Iwasaki H. Iwasaki Department of Experimental Chemotherapy, Osaka University, Japan. Search for more papers by this author M. Takahagi M. Takahagi Department of Experimental Chemotherapy, Osaka University, Japan. Search for more papers by this author T. Shiba T. Shiba Department of Experimental Chemotherapy, Osaka University, Japan. Search for more papers by this author A. Nakata A. Nakata Department of Experimental Chemotherapy, Osaka University, Japan. Search for more papers by this author H. Shinagawa H. Shinagawa Department of Experimental Chemotherapy, Osaka University, Japan. Search for more papers by this author Author Information H. Iwasaki1, M. Takahagi1, T. Shiba1, A. Nakata1 and H. Shinagawa1 1Department of Experimental Chemotherapy, Osaka University, Japan. The EMBO Journal (1991)10:4381-4389https://doi.org/10.1002/j.1460-2075.1991.tb05016.x PDFDownload PDF of article text and main figures. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Genetic evidence suggests that the Escherichia coli ruvC gene is involved in DNA repair and in the late step of RecE and RecF pathway recombination. To study the biochemical properties of RuvC protein, we overproduced and highly purified the protein. By employing model substrates, we examined the possibility that RuvC protein is an endonuclease that resolves the Holliday structure, an intermediate in genetic recombination in which two double-stranded DNA molecules are linked by single-stranded crossover. RuvC protein cleaves cruciform junctions, which are formed by the extrusion of inverted repeat sequences from a supercoiled plasmid and which are structurally analogous to Holliday junctions, by introducing nicks into strands with the same polarity. The nicked ends are ligated by E.coli or T4 DNA ligases. Analysis of the cleavage sites suggests that DNA topology rather than a particular sequence determines the cleavage site. RuvC protein also cleaves Holliday junctions which are formed between gapped circular and linear duplex DNA by the function of RecA protein. However, it does not cleave a synthetic four-way junction that does not possess homology between arms. The active form of RuvC protein, as studied by gel filtration, is a dimer. This is mechanistically suited for an endonuclease involved in swapping DNA strands at the crossover junctions. From these properties of RuvC protein and the phenotypes of the ruvC mutants, we infer that RuvC protein is an endonuclease that resolves Holliday structures in vivo. Previous ArticleNext Article Volume 10Issue 131 December 1991In this issue RelatedDetailsLoading ...