PMID: 16649659May 3, 2006Paper

Antirestriction and antimodification activities of the ArdA protein encoded by the IncI1 transmissive plasmids R-64 and ColIb-P9

Genetika
G B Zavil'gel'skiĭS M Rastorguev

Abstract

Proteins of the Ard family are specific inhibitors of type I restriction-modification enzymes. The ArdA of R64 is highly homologous to ColIb-P9 ArdA, differing only by four amino acid residues of the overall 166. However, unlike ColIb-P9 ArdA, which inhibits both the endonuclease and the methylase activities of EcoKI, the R64 ArdA protein inhibits only the endonuclease activity of this enzyme. The mutant forms of R64 ArdA--A29T, S43A, and Y75W, capable of partially reversing the protein to ColIb-P9 ArdA form--were produced by directed mutagenesis. It was demonstrated that only Y75W mutation of these three variants essentially influenced the functional activity of ArdA: the antimodification activity was restored to approximately 90-99%. It is assumed that R64 ArdA inhibits formation of the complex between unmodified DNA and the R subunit of the type I restriction-modification enzyme EcoKI (R2M2S), which translocates and cleaves DNA. ColIb-P9 ArdA protein is capable of forming the DNA complex not only with the R subunit, but also with the S subunit, which contacts sK site (containing modified adenine residues) in DNA. ArdA bound to the specific sK site inhibits concurrently the endonuclease and methylase activities of EcoKI (R2M2...Continue Reading

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Jan 1, 1985·Molecular & General Genetics : MGG·A A BelogurovG B Zavilgelsky
Jul 20, 1993·Journal of Molecular Biology·B Rost, C Sander
Jun 6, 2000·Microbiology and Molecular Biology Reviews : MMBR·N E Murray
Jan 24, 2002·Molecular Cell·M D WalkinshawD T F Dryden
Dec 5, 2003·Nucleic Acids Research·Wil A M Loenen

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Citations

Oct 10, 2007·Biochemistry. Biokhimii︠a︡·G B Zavilgelsky, S M Rastorguev
Apr 2, 2009·Molekuliarnaia biologiia·G B Zavil'gel'skiĭS M Rastorguev
May 12, 2009·Molekuliarnaia biologiia·G B Zavil'gel'skiĭ, S M Rastorguev

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