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Covalent linkage of a protein to a defined nucleotide sequence at the 5'-terminus of virion and replicative intermediate RNAs of poliovirus

Flanegan, James B. and Pettersson, Ralf F. and Ambros, Victor and Hewlett, Martinez J. and Baltimore, David (1977) Covalent linkage of a protein to a defined nucleotide sequence at the 5'-terminus of virion and replicative intermediate RNAs of poliovirus. Proceedings of the National Academy of Sciences of the United States of America, 74 (3). pp. 961-965. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:FLApnas77a

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Abstract

The 5'-terminus of poliovirus polyribosomal RNA is pUp. A candidate for the 5'-terminus of poliovirion RNA was recovered as a compound migrating toward the cathode when 32P-labeled virion RNA was completely digested with ribonucleases T1, T2, and A and analyzed by paper ionophoresis at pH 3.5. Treatment with proteinase K reversed its direction of migration, indicating the presence of protein. Treatment with venom phosphodiesterase liberated all of the radioactivity as pUp, suggesting that poliovirion RNA has a protein-pUp 5'-terminus. Treatment of virion RNA with T1 ribonuclease alone generated a proteinase K-sensitive oligoribonucleotide. Analysis of the oligoribonucleotide using ribonucleases A and U2 showed its structure to be protein-pU-U-A-A-A-A-C-A-G. Digests of replicative intermediate RNA contained sufficient protein-pUp to suggest that this structure is at the 5'-end of most nascent poliovirus RNA molecules. We suggest that a protein-nucleotide structure acts as a primer for initiating synthesis of poliovirus RNA.


Item Type:Article
Additional Information:© 1977 by the National Academy of Sciences. Contributed by David Baltimore, December 27, 1976. We are grateful to Dr. Eckard Wimmer for communicating to us results from his laboratory prior to their publication. We thank Dr. John Rose for many invaluable suggestions that greatly assisted in the completion of this work. Helpful suggestions by Dr. Naomi Guttman and the technical assistance of Kahan Smith are gratefully acknowledged. 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 A105179. R.F.P. is the recipient of a Public Health Service International Fellowship F05 TW-2245. M.J.H. was a postdoctoral fellow of the American Cancer Society. D.B. is a Research Professor of the American Cancer Society.
Subject Keywords:picornaviruses; RNA synthesis; paper ionophoresis; mRNA
Record Number:CaltechAUTHORS:FLApnas77a
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:FLApnas77a
Alternative URL:http://dx.doi.org/10.1073/pnas.74.3.961
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:7420
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:13 Feb 2007
Last Modified:26 Dec 2012 09:31

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