Structural model for the multisubunit Type IC restriction-modification DNA methyltransferase M.EcoR124I in complex with DNA.

Agnieszka Obarska; Alex Blundell; Marcin Feder; Stepánka Vejsadová; Eva Sisáková; Marie Weiserová; Janusz M Bujnicki; Keith Firman

doi:10.1093/nar/gkl132

PubMed · 16614449

Structural model for the multisubunit Type IC restriction-modification DNA methyltransferase M.EcoR124I in complex with DNA.

Agnieszka Obarska·Alex Blundell·Marcin Feder·Stepánka Vejsadová·Eva Sisáková·Marie Weiserová·Janusz M Bujnicki·Keith Firman

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Abstract

Recent publication of crystal structures for the putative DNA-binding subunits (HsdS) of the functionally uncharacterized Type I restriction-modification (R-M) enzymes MjaXIP and MgeORF438 have provided a convenient structural template for analysis of the more extensively characterized members of this interesting family of multisubunit molecular motors. Here, we present a structural model of the Type IC M.EcoR124I DNA methyltransferase (MTase), comprising the HsdS subunit, two HsdM subunits, the cofactor AdoMet and the substrate DNA molecule. The structure was obtained by docking models of individual subunits generated by fold-recognition and comparative modelling, followed by optimization of inter-subunit contacts by energy minimization. The model of M.EcoR124I has allowed identification of a number of functionally important residues that appear to be involved in DNA-binding. In addition, we have mapped onto the model the location of several new mutations of the hsdS gene of M.EcoR124I that were produced by misincorporation mutagenesis within the central conserved region of hsdS, we have mapped all previously identified DNA-binding mutants of TRD2 and produced a detailed analysis of the location of surface-modifiable lysines. The model structure, together with location of the mutant residues, provides a better background on which to study protein-protein and protein-DNA interactions in Type I R-M systems.

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Amino Acid Sequence Bacterial Proteins Conserved Sequence DNA DNA Restriction-Modification Enzymes DNA-Binding Proteins Models, Molecular Molecular Sequence Data Mutation Protein Subunits Sequence Alignment Sequence Analysis, DNA Site-Specific DNA-Methyltransferase (Adenine-Specific)Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

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Agnieszka Obarska, Alex Blundell, Marcin Feder, Stepánka Vejsadová, Eva Sisáková, Marie Weiserová, Janusz M Bujnicki, Keith Firman. 2006-04-13. Structural model for the multisubunit Type IC restriction-modification DNA methyltransferase M.EcoR124I in complex with DNA.. https://doi.org/10.1093/nar%2Fgkl132

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