Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1974 Oct;71(10):3979–3983. doi: 10.1073/pnas.71.10.3979

The 5′-Terminal Nucleotide Sequences of the Double-Stranded RNA of Human Reovirus

Kin-Ichiro Miura 1, Kumiko Watanabe 1, Masahiro Sugiura 1, Aaron J Shatkin *
PMCID: PMC434310  PMID: 4530278

Abstract

The 5′-terminal nucleotide sequences of human reovirus double-stranded RNA were determined after labeling the RNA with [32P]phosphate by polynucleotide kinase. The 5′ terminal were labeled to only a limited extent prior to sequential oxidation, β-elimination, and phosphomonoesterase treatment, indicating that the terminal phosphates were in a modified, blocked configuration. Each genome segment, after removing the blocking group, contained the same two 5′-terminal sequences: GpApUp in one chain and G*pCp in the other. G*p is a derivative of guanylic acid, probably 2′-O-methyl-Gp, which renders the 5′-terminal sequence resistant to hydrolysis by alkali. The results indicate that the transcription of reovirus double-stranded RNA strats from the 3′ end complementary to the G*pCp-terminal, resulting in the synthesis of single-stranded mRNA carrying the same 5′ sequence as the G*pCp-chain. The presence of a modified nucleotide at the 5′ terminus of the strand complementary to the mRNA template is a feature common to another double-stranded RNA virus, cytoplasmic polyhedrosis virus.

Keywords: modified 5′ termini

Full text

PDF
3980

Images in this article

3981Image
on p.3981
3981Image
on p.3981

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Banerjee A. K., Shatkin A. J. Guanosine-5'-diphosphate at the 5' termini of reovirus RNA: evidence for a segmented genome within the virion. J Mol Biol. 1971 Nov 14;61(3):643–653. doi: 10.1016/0022-2836(71)90069-6. [DOI] [PubMed] [Google Scholar]
  2. Banerjee A. K., Ward R., Shatkin A. J. Initiation of reovirus mRNA synthesis in vitro. Nat New Biol. 1971 Apr 7;230(14):169–172. doi: 10.1038/newbio230169a0. [DOI] [PubMed] [Google Scholar]
  3. Borsa J., Graham A. F. Reovirus: RNA polymerase activity in purified virions. Biochem Biophys Res Commun. 1968 Dec 30;33(6):895–901. doi: 10.1016/0006-291x(68)90396-3. [DOI] [PubMed] [Google Scholar]
  4. Fujii-Kawata I., Miura K. I. Segments of genome of viruses containing double-stranded ribonucleic acid. J Mol Biol. 1970 Jul 28;51(2):247–253. doi: 10.1016/0022-2836(70)90140-3. [DOI] [PubMed] [Google Scholar]
  5. Lewandowski L. J., Kalmakoff J., Tanada Y. Characterization of a Ribonucleic Acid Polymerase Activity Associated with Purified Cytoplasmic Polyhedrosis Virus of the Silkworm Bombyx mori. J Virol. 1969 Dec;4(6):857–865. doi: 10.1128/jvi.4.6.857-865.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Miura K., Watanabe K., Sugiura M. 5'-Terminal nucleotide sequences of the double-stranded RNA of silkworm cytoplasmic polyhedrosis virus. J Mol Biol. 1974 Jun 15;86(1):31–48. doi: 10.1016/s0022-2836(74)80005-7. [DOI] [PubMed] [Google Scholar]
  7. Nichols J. L., Hay A. J., Joklik W. K. 5'-terminal nucleotide sequence of reovirus mRNA synthesized in vitro. Nat New Biol. 1972 Jan 26;235(56):105–107. doi: 10.1038/newbio235105a0. [DOI] [PubMed] [Google Scholar]
  8. Richardson C. C. Phosphorylation of nucleic acid by an enzyme from T4 bacteriophage-infected Escherichia coli. Proc Natl Acad Sci U S A. 1965 Jul;54(1):158–165. doi: 10.1073/pnas.54.1.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Richardson C. C. The 5'-terminal nucleotides of T7 bacteriophage deoxyribonucleic acid. J Mol Biol. 1966 Jan;15(1):49–61. doi: 10.1016/s0022-2836(66)80208-5. [DOI] [PubMed] [Google Scholar]
  10. Schonberg M., Silverstein S. C., Levin D. H., Acs G. Asynchronous synthesis of the complementary strands of the reovirus genome. Proc Natl Acad Sci U S A. 1971 Feb;68(2):505–508. doi: 10.1073/pnas.68.2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Shatkin A. J. Methylated messenger RNA synthesis in vitro by purified reovirus. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3204–3207. doi: 10.1073/pnas.71.8.3204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Shatkin A. J., Sipe J. D., Loh P. Separation of ten reovirus genome segments by polyacrylamide gel electrophoresis. J Virol. 1968 Oct;2(10):986–991. doi: 10.1128/jvi.2.10.986-991.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Shatkin A. J., Sipe J. D. RNA polymerase activity in purified reoviruses. Proc Natl Acad Sci U S A. 1968 Dec;61(4):1462–1469. doi: 10.1073/pnas.61.4.1462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Shimotono K., Miura K. 5'-Terminal structure of messenger RNA transcribed by RNA polymerase of silkworm cytoplasmic polyhedrosis virus containing double-stranded RNA. J Mol Biol. 1974 Jun 15;86(1):21–30. doi: 10.1016/s0022-2836(74)80004-5. [DOI] [PubMed] [Google Scholar]
  15. Shimotono K., Miura K. Single-stranded RNA synthesis in vitro by the RNA polymerase associated with cytoplasmic polyhedrosis virus containing double-stranded RNA. J Biochem. 1973 Jul;74(1):117–125. doi: 10.1093/oxfordjournals.jbchem.a130214. [DOI] [PubMed] [Google Scholar]
  16. Shimotono K., Miura K. Transcription of double-stranded RNA in cytoplasmic polyhedrosis virus in vtro. Virology. 1973 May;53(1):283–286. [PubMed] [Google Scholar]
  17. Takanami M. Analysis of the 5'-terminal nucleotide sequences of ribonucleic acids 1. the 5'-termini of Excherichia coli ribosomal RNA. J Mol Biol. 1967 Jan 28;23(2):135–148. doi: 10.1016/s0022-2836(67)80022-6. [DOI] [PubMed] [Google Scholar]
  18. Takanami M. [Structure of the 5'-terminal of high-polymer nucleic acids--analytical method and its application]. Seikagaku. 1967 Aug;39(8):455–462. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES