The Orf18 Gene Product from Conjugative Transposon Tn916 Is an ArdA Antirestriction Protein that Inhibits Type I DNA Restriction-Modification Systems

Serfiotis-Mitsa, D., Roberts, G. A., Cooper, L. P., White, J. H., Nutley, M., Cooper, A., Blakely, G. W. and Dryden, D. T. F. (2008) The Orf18 Gene Product from Conjugative Transposon Tn916 Is an ArdA Antirestriction Protein that Inhibits Type I DNA Restriction-Modification Systems. Journal of Molecular Biology, 383(5), pp. 970-981. (doi:10.1016/j.jmb.2008.06.005)

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Abstract

Gene orf18, which is situated within the intercellular transposition region of the conjugative transposon Tn916 from the bacterial pathogen Enterococcus faecalis, encodes a putative ArdA (alleviation of restriction of DNA A) protein. Conjugative transposons are generally resistant to DNA restriction upon transfer to a new host. ArdA from Tn916 may be responsible for the apparent immunity of the transposon to DNA restriction and modification (R/M) systems and for ensuring that the transposon has a broad host range. The orf18 gene was engineered for overexpression in Escherichia coli, and the recombinant ArdA protein was purified to homogeneity. The protein appears to exist as a dimer at nanomolar concentrations but can form larger assemblies at micromolar concentrations. R/M assays revealed that ArdA can efficiently inhibit R/M by all four major classes of Type I R/M enzymes both in vivo and in vitro. These R/M systems are present in over 50% of sequenced prokaryotic genomes. Our results suggest that ArdA can overcome the restriction barrier following conjugation and so helps increase the spread of antibiotic resistance genes by horizontal gene transfer..

Item Type:Articles
Keywords:antirestriction ANTIRESTRICTION PROTEIN Ard protein ASSEMBLIES BACILLUS-SUBTILIS BACTERIOPHAGE T7 conjugation DNA ECOKI ENTEROCOCCUS-FAECALIS ENZYME ESCHERICHIA-COLI HELP IN-VITRO IN-VIVO METHYLTRANSFERASE OVEREXPRESSION PATHOGEN PLASMID PKM101 PROTEIN RESISTANCE RESTRICTION/MODIFICATION ENZYME SECONDARY STRUCTURE SYSTEM SYSTEMS Tn916 transposon
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Nutley, Mrs Margaret and Cooper, Professor Alan
Authors: Serfiotis-Mitsa, D., Roberts, G. A., Cooper, L. P., White, J. H., Nutley, M., Cooper, A., Blakely, G. W., and Dryden, D. T. F.
College/School:College of Science and Engineering > School of Chemistry
Journal Name:Journal of Molecular Biology
ISSN:0022-2836

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