Flanegan, James B. and Baltimore, David (1977) Poliovirus-specific primer-dependent RNA polymerase able to copy poly(A). Proceedings of the National Academy of Sciences of the United States of America, 74 (9). pp. 3677-3680. ISSN 0027-8424 http://resolver.caltech.edu/CaltechAUTHORS:FLApnas77b
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A template-dependent RNA polymerase has been isolated from poliovirus-infected cells by assaying for the ability of the enzyme to copy poly(A) complexed to an oligo(U) primer. The polymerase was solubilized with detergent, and RNA was removed by precipitation with 2 M LiCl. The solubilized polymerase required both poly(A) and oligo(U) for activity and was stimulated by Mg2+ but was inhibited by Mn2+. Poly(A)· oligo(U)-dependent poly(U) polymerase was not found in extracts of HeLa cells until about 2 hr after poliovirus infection, and then there was a linear increase in activity until about 5 hr. Analysis of the polymerase by glycerol gradient centrifugation showed that the majority of the activity sedimented at about 4 S, indicating that it was no longer complexed with high-molecular-weight RNA or cellular membranes. This poly(A)· oligo(U)-dependent polymerase activity could represent an important component of the poliovirus RNA-dependent RNA polymerase.
|Additional Information:||© 1977 by the National Academy of Sciences. Contributed by David Baltimore, June 9, 1977. We gratefully acknowledge the helpful suggestions of Dr. Ralf Pettersson and Dr. John Rose and the excellent technical assistance of Kahan Smith and Victor Ambros. This work was supported by Grants AI08388, CA 12174, and CA 14051 from the National Institutes of Health. J.B.F. is the recipient of a National Institutes of Health Postdoctoral Fellowship F32 AI05179. D.B. is a Research Professor of the American Cancer Society. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.|
|Subject Keywords:||RNA-dependent RNA polymerase (replicase); picornaviruses|
|Usage Policy:||No commercial reproduction, distribution, display or performance rights in this work are provided.|
|Deposited By:||Tony Diaz|
|Deposited On:||13 Feb 2007|
|Last Modified:||26 Dec 2012 09:31|
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