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. 1978 Aug;27(2):357–365. doi: 10.1128/jvi.27.2.357-365.1978

Identification of a protein linked to nascent poliovirus RNA and to the polyuridylic acid of negative-strand RNA.

R F Pettersson, V Ambros, D Baltimore
PMCID: PMC354174  PMID: 211265

Abstract

A protein similar to that previously demonstrated on poliovirus RNA and replicative intermediate RNA (VPg) was found on all sizes of nascent viral RNA molecules and on the polyuridylic acid isolated from negative-strand RNA. 32P-labeled nascent chains were released from their template RNA and fractionated by exclusion chromatography on agarose. Fingerprint analysis using two-dimensional polyacrylamide gels of RNase T1 oligonucleotides derived from nascent chains of different lengths showed that a size fractionation of nascent chains was achieved. VPg was recovered from nascent chains varying in length from 7,500 nucleotides (full-sized RNA) to about 500 nucleotides. No other type of 5' terminus could be demonstrated on nascent RNA, and the yield of VPg was consistent with one molecule of the protein on each nascent chain. These results are consistent with the concept that the protein is added to the 5' end of the growing RNA chains at a very early stage, possibly as a primer of RNA synthesis. Analysis of the polyuridylic acid tract isolated from the replicative intermediate and double-stranded RNAs indicated that a protein of the same size as that found on the nascent chains and virion RNA is also linked to the negative-strand RNAs. It is likely that a similar mechanism is responsible for initiation of synthesis of both plus- and minus-strand RNAs.

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Selected References

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  1. Baltimore D., Girard M. An intermediate in the synthesis of poliovirus RNA. Proc Natl Acad Sci U S A. 1966 Aug;56(2):741–748. doi: 10.1073/pnas.56.2.741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baltimore D., Girard M., Darnell J. E. Aspects of the synthesis of poliovirus RNA and the formation of virus particles. Virology. 1966 Jun;29(2):179–189. doi: 10.1016/0042-6822(66)90024-9. [DOI] [PubMed] [Google Scholar]
  3. Baltimore D. Structure of the poliovirus replicative intermediate RNA. J Mol Biol. 1968 Mar 14;32(2):359–368. doi: 10.1016/0022-2836(68)90015-6. [DOI] [PubMed] [Google Scholar]
  4. Cole C. N., Baltimore D. Defective interfering particles of poliovirus. II. Nature of the defect. J Mol Biol. 1973 May 25;76(3):325–343. doi: 10.1016/0022-2836(73)90508-1. [DOI] [PubMed] [Google Scholar]
  5. Flanegan J. B., Petterson R. F., Ambros V., Hewlett N. J., Baltimore D. Covalent linkage of a protein to a defined nucleotide sequence at the 5'-terminus of virion and replicative intermediate RNAs of poliovirus. Proc Natl Acad Sci U S A. 1977 Mar;74(3):961–965. doi: 10.1073/pnas.74.3.961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hewlett M. J., Rose J. K., Baltimore D. 5'-terminal structure of poliovirus polyribosomal RNA is pUp. Proc Natl Acad Sci U S A. 1976 Feb;73(2):327–330. doi: 10.1073/pnas.73.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Lee Y. F., Nomoto A., Detjen B. M., Wimmer E. A protein covalently linked to poliovirus genome RNA. Proc Natl Acad Sci U S A. 1977 Jan;74(1):59–63. doi: 10.1073/pnas.74.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lee Y. F., Wimmer E. "Fingerprinting" high molecular weight RNA by two-dimensional gel electrophoresis: application to poliovirus RNA. Nucleic Acids Res. 1976 Jul;3(7):1647–1658. doi: 10.1093/nar/3.7.1647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nomoto A., Detjen B., Pozzatti R., Wimmer E. The location of the polio genome protein in viral RNAs and its implication for RNA synthesis. Nature. 1977 Jul 21;268(5617):208–213. doi: 10.1038/268208a0. [DOI] [PubMed] [Google Scholar]
  11. Pettersson R. F., Flanegan J. B., Rose J. K., Baltimore D. 5'-Terminal nucleotide sequences of polio virus polyribosomal RNA and virion RNA are identical. Nature. 1977 Jul 21;268(5617):270–272. doi: 10.1038/268270a0. [DOI] [PubMed] [Google Scholar]
  12. Pettersson R. F., Hewlett M. J., Baltimore D., Coffin J. M. The genome of Uukuniemi virus consists of three unique RNA segments. Cell. 1977 May;11(1):51–63. doi: 10.1016/0092-8674(77)90316-6. [DOI] [PubMed] [Google Scholar]
  13. Sangar D. V., Rowlands D. J., Harris T. J., Brown F. Protein covalently linked to foot-and-mouth disease virus RNA. Nature. 1977 Aug 18;268(5621):648–650. doi: 10.1038/268648a0. [DOI] [PubMed] [Google Scholar]
  14. Spector D. H., Baltimore D. Polyadenylic acid on poliovirus RNA IV. Poly(U) in replicative intermediate and double-stranded RNA. Virology. 1975 Oct;67(2):498–505. doi: 10.1016/0042-6822(75)90450-x. [DOI] [PubMed] [Google Scholar]
  15. Spector D. H., Baltimore D. Polyadenylic acid on poliovirus RNA. II. poly(A) on intracellular RNAs. J Virol. 1975 Jun;15(6):1418–1431. doi: 10.1128/jvi.15.6.1418-1431.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Yogo Y., Teng M. H., Wimmer E. Poly(U) in poliovirus minus RNA is 5'-terminal. Biochem Biophys Res Commun. 1974 Dec 23;61(4):1101–1109. doi: 10.1016/s0006-291x(74)80397-9. [DOI] [PubMed] [Google Scholar]
  17. Yogo Y., Wimmer E. Poly (A) and poly (U) in poliovirus double stranded RNA. Nat New Biol. 1973 Apr 11;242(119):171–174. doi: 10.1038/newbio242171a0. [DOI] [PubMed] [Google Scholar]
  18. de Wachter R., Fiers W. Preparative two-dimensional polyacrylamide gel electrophoresis of 32 P-labeled RNA. Anal Biochem. 1972 Sep;49(1):184–197. doi: 10.1016/0003-2697(72)90257-6. [DOI] [PubMed] [Google Scholar]

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