Effects of poliovirus 3AB protein on 3D polymerase-catalyzed reaction.
Poliovirus RNA replication requires the activities of a viral RNA-dependent RNA polymerase, 3Dpol, in conjunction with several additional viral and likely cellular proteins. The importance of both the 3A and 3B coding regions has been documented previously by genetic tests, and their biochemical activities have been the subject of several recent investigations. In this study, we examined the previously reported stimulation of 3D-catalyzed RNA synthesis by 3AB. We show that 3AB does not stimulate RNA synthesis on templates that are stably base paired to a primer, indicating that 3AB does not stabilize or otherwise activate 3Dpol for chain elongation. Similarly, it does not alter the kinetic parameters or binding affinities of 3D for substrates. In the absence of a primer, or in the presence of a primer that does not form a stable hybrid with the template, 3AB increases the utilization of 3'-hydroxyl termini as sites for chain elongation by 3D, and thereby stimulates RNA synthesis. 3AB may interact with and stabilize these sites and/or may recruit 3Dpol to the site, resulting in stimulation of the initiation of elongation events. We propose that this activity is required for stabilizing weak interactions that occur during nucleotidyl-protein-primed initiation events in the viral RNA replication complex.
Richards
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Department of Molecular Biology and Biochemistry, University of California, Irvine, California 92697, USA.
Ehrenfeld
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D001483:Base Sequence; D002384:Catalysis; D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D012324:RNA-Dependent RNA Polymerase; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D014758:Viral Core Proteins / Q000378:metabolism*
D017931:DNA Primers; D011994:Recombinant Proteins; D014758:Viral Core Proteins; C090443:polymerase 3Dpol, poliovirus; D012324:RNA-Dependent RNA Polymerase; D012321:DNA-Directed RNA Polymerases