Sse9I restriction-modification system: Organization of genes and structural comparison of proteins |
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| Authors: | D A Gonchar M A Abdurashitov S S Okhapkina D A Shagin E V Kileva S Kh Degtyarev |
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| Institution: | (1) Institute of Molecular Biology and Biophysics, Siberian Division, Russian Academy of Medical Sciences, Novosibirsk, 630117, Russia;(2) SibEnzyme Research and Production Association, Novosibirsk, 630117, Russia;(3) Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia |
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| Abstract: | The nucleotide sequence was established for the operon of the Sse9I type II restriction-modification system of Sporosarcina species 9D. The enzymes of the Sse9I system recognize the 5′-AATT-3′ tetranucleotide. The operon includes three genes, sse9IC-sse9IR-sse9IM, which are transcribed unidirectionally and code, respectively, for the controller protein (C.Sse9I), restriction endonuclease (R.Sse9I), and DNA methyltransferase (M.Sse9I). The region immediately upstream of sse9IC was found to contain a conserved nucleotide sequence (C box) providing a binding site for C. Sse9I. The amino acid sequences of C.Sse9I and R.Sse9I were compared with those of related proteins. In the case of R.Sse9I, the highest homology was observed with the R.MunI (5′-CAATTG-3′) and R.EcoRI (5′-GAATTC-3′) regions that harbor the amino acid residues involved in recognizing the AATT inner tetranucleotide. The sse9IR gene was cloned in an expression vector, and recombinant R.Sse 9I was isolated. |
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| Keywords: | restriction-modification system Sporosarcina species 9D restriction endonuclease C-protein chromosome walking cloning expression |
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