Single-stranded DNA binding and methylation by EcoP1I DNA methyltransferase |
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Authors: | Sistla Srivani Krishnamurthy Vinita Rao Desirazu N |
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Affiliation: | Department of Biochemistry, Indian Institute of Science, Bangalore, Karnataka, India. |
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Abstract: | EcoP1I methyltransferase (M.EcoP1I) belongs to the type III restriction-modification system encoded by prophage P1 that infects Escherichia coli. Binding of M.EcoP1I to double-stranded DNA and single-stranded DNA has been characterized. Binding to both single- and double-stranded DNA could be competed out by unlabeled single-stranded DNA. Metal ions did not influence DNA binding. Interestingly, M.EcoP1I was able to methylate single-stranded DNA. Kinetic parameters were determined for single- and double-stranded DNA methylation. This feature of the enzyme probably functions in protecting the phage genome from restriction by type III restriction enzymes and thus could be considered as an anti-restriction system. This study describing in vitro methylation of single-stranded DNA by the type III methyltransferase EcoP1I allows understanding of the mechanism of action of these enzymes and also their role in the biology of single-stranded phages. |
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Keywords: | Anti-restriction Binding Methylation single-stranded DNA |
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