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Vol 43(2009) N 2 p. 241-248;
G. B. Zavilgelsky, S. M. Rastorguev

Antirestriction proteins ArdA and Ocr as efficient inhibitors of type I restriction-modification enzymes

State Research Center GosNIIgenetika Moscow, Moscow, 117545, Russia
Received - 2008-07-08; Accepted - 2008-07-08

Genes encoding antirestriction proteins are found in transmissble plasmids (ardABC) and bacteriophage genomes (ocr, darA). Antirestriction proteins inhibit type I restriction-modification enzymes and thus protect the unmodified plasmid or phage DNA from degradation. Antirestriction proteins belong to the family of DNA-mimicry proteins, whose spatial structure mimics the B-form of DNA. Based on an analysis of the mutant forms of ArdA and Ocr obtained by site-directed mutagenesis and the native form of ArdA that specifically inhibit type I restriction enzymes but do not affect their methylase activity, a model is proposed to describe the complex formation between an antirestriction protein and a type I restriction-modification enzyme (R2M2S): antirestriction proteins can displace a DNA strand from its binding sites in the S subunit (which contacts a specific site on DNA) and in the R subunit (which translocates the DNA strand and cleaves it). Antirestriction and antimodification activities of ArdA and Ocr as a function of ardA and ocr expression levels were studied by cloning the genes under a strictly regulated promoter.

type I restriction-modification enzymes, antirestriction, ArdA, Ocr, transmissible plasmid, bacteriophage, DNA mimicry



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