Investigation of Activity of Recombinant Mengovirus RNA-Dependent RNA Polymerase and Its Mutants

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G. S. Shatskaya^1*, V. L. Drutsa¹, O. N. Koroleva², I. A. Osterman², and T. M. Dmitrieva¹

¹Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119992 Moscow, Russia; fax: (495) 939-3181; E-mail: barsa19@rambler.ru; drutsa@genebee.msu.su

²Faculty of Chemistry, Lomonosov Moscow State University, 119992 Moscow, Russia; fax: (495) 939-3181; E-mail: koroleva@genebee.msu.su; osterman@yandex.ru

^* To whom correspondence should be addressed.

Received September 18, 2012; Revision received October 2, 2012

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The activities of wild-type mengovirus RNA polymerase (RdRP) and of its three mutants with C-terminal tryptophan residue replaced by residues of alanine (W460A), phenylalanine (W460F), or tyrosine (W460Y) were studied. The proteins were expressed in E. coli and purified by affinity chromatography with the IMPACT system. The isolated recombinant proteins were studied using a cell-free replication system on elongation of oligo(U) primer on RNA template corresponding to the 3′-terminal 366-meric fragment of the mengovirus RNA. The activities of the mutant polymerases were comparable to that of the wild-type enzyme.

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KEY WORDS: RNA-dependent RNA polymerase, mengovirus, IMPACT system

DOI: 10.1134/S0006297913010124

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