Affinity photo-crosslinking study of the DNA base flipping pathway by HhaI methyltransferase | Zendy

D. Daujotyte | Zendy; Saulius Klimašauskas | Zendy

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Affinity photo-crosslinking study of the DNA base flipping pathway by HhaI methyltransferase

Author(s) -

D. Daujotyte,

Saulius Klimašauskas

Publication year - 2000

Publication title -

nucleic acids symposium series

Language(s) - English

Resource type - Journals

eISSN - 1746-8272

pISSN - 0261-3166

DOI - 10.1093/nass/44.1.271

Subject(s) - dna , cytosine , base pair , methyltransferase , recognition sequence , dna methyltransferase , methylation , biophysics , dodecameric protein , covalent bond , chemistry , biochemistry , restriction enzyme , biology , organic chemistry

HhaI DNA methyltransferase flips the inner cytosine in the recognition sequence 5'-GCGC-3' out of the DNA helix and into the catalytic site for methylation. To identify intermediate states on the base-flipping pathway, affinity photo-crosslinking experiments were performed with synthetic dodecamer duplexes containing modified bases 2-thiothymine (2sT) or 4-thiothymine (4sT) at the target base position. Here we show that the DNA strand containing 2sT, but not 4sT, covalently cross-links to the HhaI methyltransferase upon irradiation at 340-360 nm.

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