Please use this identifier to cite or link to this item: https://scholarbank.nus.edu.sg/handle/10635/112017
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dc.titlePoint mutations which drastically affect the polymerization activity of encephalomyocarditis virus RNA-dependent RNA polymerase correspond to the active site of Eacherichia coli DNA polymerase I
dc.contributor.authorSankar, S.
dc.contributor.authorPorter, A.G.
dc.date.accessioned2014-11-28T02:52:14Z
dc.date.available2014-11-28T02:52:14Z
dc.date.issued1992-05-15
dc.identifier.citationSankar, S.,Porter, A.G. (1992-05-15). Point mutations which drastically affect the polymerization activity of encephalomyocarditis virus RNA-dependent RNA polymerase correspond to the active site of Eacherichia coli DNA polymerase I. Journal of Biological Chemistry 267 (14) : 10168-10176. ScholarBank@NUS Repository.
dc.identifier.issn00219258
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/112017
dc.description.abstractThe inhibitor sensitivity and functional domains of recombinant encephalomyocarditis (EMC) virus RNA-dependent RNA polymerase (3Dpol) have been extensively analyzed. The inhibitor profiles of EMC virus 3Dpol and Escherichia coli DNA-dependent RNA polymerase are distinct, and experiments with substrate analogs indicate that EMC virus 3Dpol lacks reverse transcriptase activity. Twenty amino acid substitutions were engineered in EMC virus 3Dpol based on sequence alignments of viral RNA-dependent RNA polymerases that identified conserved amino acid residues within motifs. Ten out of 17 conservative substitutions within the four most conserved motifs reduced the RNA polymerase activity of the mutants to 0-6% of the activity of the wild-type enzyme, demonstrating the importance of these amino acids in the structure and/or function of EMC virus 3Dpol. Remarkably, 5 of the 10 mutations in EMC virus 3Dpol which had the most drastic effect on its RNA polymerase activity (D240E, S293T, N302Q, G332A, and D333E) were found to correspond to active site residues in E. coli DNA-dependent DNA polymerase I (Klenow). Our results reveal that a basic structural and functional framework is conserved in the most distantly related classes of nucleic acid polymerases and demonstrate the validity of modeling the active site of an RNA-dependent RNA polymerase on the known structure of a DNA polymerase.
dc.sourceScopus
dc.typeArticle
dc.contributor.departmentINSTITUTE OF MOLECULAR & CELL BIOLOGY
dc.description.sourcetitleJournal of Biological Chemistry
dc.description.volume267
dc.description.issue14
dc.description.page10168-10176
dc.description.codenJBCHA
dc.identifier.isiutNOT_IN_WOS
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