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T J Su, M R Tock, S U Egelhaaf, W C Poon, and D T Dryden (2005)

Dna bending by m.ecoki methyltransferase is coupled to nucleotide flipping

Nucleic Acids Research 33:3235–3244.

The maintenance methyltransferase M.EcoKI recognizes the bipartite DNA sequence 5'-A (A) under bar CNNN-NNNG (T) under bar GC-3', where N is any nucleotide. M.EcoKI preferentially methylates a sequence already containing a methylated adenine at or complementary to the underlined bases in the sequence. We find that the introduction of a single-stranded gap in the middle of the non-specific spacer, of up to 4 nt in length, does not reduce the binding affinity of M.EcoKI despite the removal of non-sequence-specific contacts between the protein and the DNA phosphate backbone. Surprisingly, binding affinity is enhanced in a manner predicted by simple polymer models of DNA flexibility. However, the activity of the enzyme declines to zero once the single-stranded region reaches 4 nt in length. This indicates that the recognition of methylation of the DNA is communicated between the two