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PMID: 198796 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Poliovirus-specific primer-dependent RNA polymerase able to copy poly(A).

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 74 ·No. 9 ·1977-09-00 ·Pages 3677-80

Flanegan JB, Baltimore D

Abstract

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.

MeSH Terms
HeLa Cells/enzymology Kinetics Nucleotidyltransferases/isolation & purification,metabolism Oligoribonucleotides Poliovirus/enzymology Poly A/pharmacology Poly U Ribonucleotides/analysis Templates, Genetic Uracil Nucleotides
Chemicals
Oligoribonucleotides Ribonucleotides Uracil Nucleotides Poly A Poly U Nucleotidyltransferases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Flanegan J B
Baltimore D
References (20)
20 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.74.9.3677
Published
1977-09-00
Pages
3677-80
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC431685
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.74.9.3677 PMC Full Text View journal details
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