PMID: 19916926Nov 18, 2009Paper

Structure-function analysis of mutant RNA-dependent RNA polymerase complexes with VPg

Biochemistry. Biokhimii︠a︡
Chaojiang GuCongyi Zheng

Abstract

The replication of the foot-and-mouth disease virus (FMDV) genome is critically dependent upon the activity of a virally encoded RNA-dependent RNA polymerase (RdRp). In this study, four mutant RdRps of FMDV were isolated from viral quasi-species treated with ribavirin, of which two were single mutants (L123F and T381A) and two were double mutants (T291I/T381I and L123F/F244L). The mutant proteins were expressed in Escherichia coli and purified by His-bind resin chromatography. In combination with real-time RT-PCR, an in vitro RNA replication system that uses genome RNA/VPg as template-primers was used to determine polymerase activity. Mutant L123F exhibited a 0.6-fold decrease (p < 0.001) in polymerase activity relative to wild-type RdRp, whereas the activity of L123F/F244L and T381A was undetectable. Surprisingly, the activity of T291I/T381I yielded a 0.7-fold increase (p < 0.001) as compared to wild-type. In order to study the structure-function relationship of RdRp, all structures of the RdRp-RNA template-primer complex were obtained through homology modeling and molecular docking. The VPg1 orientation in the RdRp-VPg1 complexes was determined and analyzed with mathematical methods. Our results reveal that the orientation of...Continue Reading

References

Apr 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·L ChicheP A Kollman
Jan 1, 1996·Journal of Molecular Recognition : JMR·D S GoodsellA J Olson
Aug 1, 1997·Protein Engineering·B A RevaA J Olson
Sep 10, 2002·Bioinformatics·Christophe LambertEric Depiereux
Jun 26, 2003·Nucleic Acids Research·Torsten SchwedeManuel C Peitsch
Jun 26, 2004·The Journal of Biological Chemistry·Pietri Puustinen, Kristiina Mäkinen
Aug 6, 2004·The Journal of Biological Chemistry·Cristina Ferrer-OrtaNuria Verdaguer
May 10, 2005·The Journal of Biological Chemistry·Jamie J ArnoldCraig E Cameron
Jun 24, 2005·Acta Crystallographica. Section A, Foundations of Crystallography·Douglas L Theobald
Sep 17, 2005·Science·Philip BradleyDavid Baker
Feb 4, 2006·The EMBO Journal·Cristina Ferrer-OrtaNuria Verdaguer
Feb 10, 2006·Journal of Biochemistry and Molecular Biology·Chao-jiang GuSan-fu Qu
May 23, 2007·Proceedings of the National Academy of Sciences of the United States of America·Cristina Ferrer-OrtaNuria Verdaguer
Apr 29, 2009·Journal of Computational Chemistry·Garrett M MorrisArthur J Olson
Nov 1, 1981·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·G Wenzel, H Uhrig

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Citations

May 6, 2015·Proceedings of the National Academy of Sciences of the United States of America·Luciana E LeopoldShawn Ahmed
Mar 21, 2020·Frontiers in Microbiology·Hailong WangChao Shen

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