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

James E. N. Taylor; P. Callow; Anna Swiderska; Geoff Kneale

doi:10.1016/j.jmb.2010.03.008

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

James E. N. Taylor, P. Callow, Anna Swiderska, Geoff Kneale

Research output: Contribution to journal › Article › peer-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 language	English
Pages (from-to)	391-399
Number of pages	9
Journal	Journal of Molecular Biology
Volume	398
Issue number	3
DOIs	https://doi.org/10.1016/j.jmb.2010.03.008
Publication status	Published - May 2010

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title = "Structural and functional analysis of the engineered type I DNA methyltransferase EcoR124I(NT)",

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.",

author = "Taylor, {James E. N.} and P. Callow and Anna Swiderska and Geoff Kneale",

year = "2010",

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language = "English",

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journal = "Journal of Molecular Biology",

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TY - JOUR

T1 - Structural and functional analysis of the engineered type I DNA methyltransferase EcoR124I(NT)

AU - Taylor, James E. N.

AU - Callow, P.

AU - Swiderska, Anna

AU - Kneale, Geoff

PY - 2010/5

Y1 - 2010/5

N2 - 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.

AB - 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.

U2 - 10.1016/j.jmb.2010.03.008

DO - 10.1016/j.jmb.2010.03.008

M3 - Article

SN - 0022-2836

VL - 398

SP - 391

EP - 399

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

IS - 3

ER -

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

Abstract

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