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In Vitro Copying of Viral Positive Strand RNA by Poliovirus Replicase. Characterization of the reaction and its products

Baron, Margaret H. and Baltimore, David (1982) In Vitro Copying of Viral Positive Strand RNA by Poliovirus Replicase. Characterization of the reaction and its products. Journal of Biological Chemistry, 257 (20). pp. 12359-12366. ISSN 0021-9258.

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Poliovirus replicase can be isolated in a form which depends on either oligo(U) or on a host cell protein for the initiation of copying of poliovirion (plus strand) RNA. The product of replicase reactions--initiated either with host factor or with oligo(U)--includes full length (35 S) RNA molecules, largely in double-stranded form, which contain the ribonuclease T1-resistant oligonucleotides of the poliovirus minus strand. For the oligo(U)-stimulated reaction, it is shown that the oligo(U) primer is covalently associated with full length product at its 5'-end. For either the host factor- or oligo(U)-dependent reactions, full length molecules appear only after 15 min of synthesis. The fraction of 35 S product is increased by raising the concentration of the limiting nucleoside triphosphate. The reaction is inhibited by as little as 100 mM salt, although it is stimulated by low (20 mM) salt concentrations. Zinc stimulates overall synthesis, but not the rate of appearance of full length molecules; the reaction is inhibited by agents which chelate zinc. Although synthesis of full length products occurs much more slowly than in the infected cell, this soluble system appears to mimic quite faithfully the initial steps of poliovirus replication.

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Baltimore, David0000-0001-8723-8190
Additional Information:© 1982 American Society for Biochemistry and Molecular Biology. Received for publication, February 2, 1981. This research was supported by grants from the National Institute of Allergy and Infectious Diseases and from the National Cancer Institute. Predoctoral scholar of the Insurance Medical Scientist Scholarship Fund and fellow of the Danforth Foundation. American Cancer Society Professor of Microbiology. 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. Section 1734 solely to indicate this fact.
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National Institute of Allergy and Infectious DiseasesUNSPECIFIED
National Cancer InstituteUNSPECIFIED
Danforth FoundationUNSPECIFIED
Issue or Number:20
Record Number:CaltechAUTHORS:20120717-091003406
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Official Citation:In vitro copying of viral positive strand RNA by poliovirus replicase. Characterization of the reaction and its products. M H Baron and D Baltimore J. Biol. Chem. 1982 257: 12359-12366.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:32499
Deposited By: Ruth Sustaita
Deposited On:17 Jul 2012 18:13
Last Modified:03 Oct 2019 04:01

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