Structural and functional analysis of the engineered type I DNA methyltransferase EcoR124I(NT)

J. Taylor, P. Callow, Anna Swiderska, Geoff Kneale

Research output: Contribution to journalArticlepeer-review

Abstract

The Type I R-M system EcoR124I is encoded by three genes. HsdM is responsible for modification (DNA methylation), HsdS for DNA sequence specificity and HsdR for restriction endonuclease activity. The trimeric methyltransferase (M(2)S) recognises the asymmetric sequence (GAAN(6)RTCG). An engineered R-M system, denoted EcoR124I(NT), has two copies of the N-terminal domain of the HsdS subunit of EcoR124I, instead of a single S subunit with two domains, and recognises the symmetrical sequence GAAN(7)TTC. We investigate the methyltransferase activity of EcoR124I(NT), characterise the enzyme and its subunits by analytical ultracentrifugation and obtain low-resolution structural models from small-angle neutron scattering experiments using contrast variation and selective deuteration of subunits.
Original languageEnglish
Pages (from-to)391-399
Number of pages9
JournalJournal of Molecular Biology
Volume398
Issue number3
DOIs
Publication statusPublished - May 2010

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