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. 1980 Jan 25;8(2):337–350. doi: 10.1093/nar/8.2.337

Assignment of reovirus mRNA ribosome binding sites to virion genome segments by nucleotide sequence analyses.

E Darzynkiewicz, A J Shatkin
PMCID: PMC327270  PMID: 7422544

Abstract

All ten reovirus genome RNA segments were radiolabeled at their 3'-termini by incubation with RNA ligase and 32pCp. The extent of radiolabeling was similar for each of the double-stranded RNAs in the genome segment mixture. Radioactivity was equally distributed between the separated plus and minus strands indicating that the 5'-cap in plus strands did not block 3'-end-labeling of minus strands. The 3'-termini of the four S and three M segments included the common sequences: ...U-A-G-C in minus strands and ...U-C-A-U-C in plus strands. By comparing the minus strand 3'-sequences with 5'-sequences of reovirus mRNAs, small-size genome segments S2, S3 and S4 were correlated with the previously sequenced initiation fragments s46, s45 and s54 derived from small class mRNAs. Medium-size genome segments M1, M2 and M3 similarly were correlated with fragments m30, m52 and m44, respectively. The N-terminal amino acid sequences deduced from the mRNA nucleotide sequences can now be assigned to the nascent chains of particular reovirus proteins.

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Selected References

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  1. 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]
  2. Both G. W., Banerjee A. K., Shatkin A. J. Methylation-dependent translation of viral messenger RNAs in vitro. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1189–1193. doi: 10.1073/pnas.72.3.1189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Both G. W., Lavi S., Shatkin A. J. Synthesis of all the gene products of the reovirus genome in vivo and in vitro. Cell. 1975 Feb;4(2):173–180. doi: 10.1016/0092-8674(75)90124-5. [DOI] [PubMed] [Google Scholar]
  4. Donis-Keller H., Maxam A. M., Gilbert W. Mapping adenines, guanines, and pyrimidines in RNA. Nucleic Acids Res. 1977 Aug;4(8):2527–2538. doi: 10.1093/nar/4.8.2527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. England T. E., Uhlenbeck O. C. 3'-terminal labelling of RNA with T4 RNA ligase. Nature. 1978 Oct 12;275(5680):560–561. doi: 10.1038/275560a0. [DOI] [PubMed] [Google Scholar]
  6. Franklin R. M. Purification and properties of the replicative intermediate of the RNA bacteriophage R17. Proc Natl Acad Sci U S A. 1966 Jun;55(6):1504–1511. doi: 10.1073/pnas.55.6.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Furuichi Y., Morgan M., Muthukrishnan S., Shatkin A. J. Reovirus messenger RNA contains a methylated, blocked 5'-terminal structure: m-7G(5')ppp(5')G-MpCp-. Proc Natl Acad Sci U S A. 1975 Jan;72(1):362–366. doi: 10.1073/pnas.72.1.362. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Furuichi Y., Muthukrishnan S., Shatkin A. J. 5'-Terminal m-7G(5')ppp(5')G-m-p in vivo: identification in reovirus genome RNA. Proc Natl Acad Sci U S A. 1975 Feb;72(2):742–745. doi: 10.1073/pnas.72.2.742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Furuichi Y., Muthukrishnan S., Tomasz J., Shatkin A. J. Mechanism of formation of reovirus mRNA 5'-terminal blocked and methylated sequence, m7GpppGmpC. J Biol Chem. 1976 Aug 25;251(16):5043–5053. [PubMed] [Google Scholar]
  10. Gomatos P. J., Tamm I. THE SECONDARY STRUCTURE OF REOVIRUS RNA. Proc Natl Acad Sci U S A. 1963 May;49(5):707–714. doi: 10.1073/pnas.49.5.707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hastings K. E., Millward S. Nucleotide sequences at the 5' termini of reovirus mRNA's. J Virol. 1978 Nov;28(2):490–498. doi: 10.1128/jvi.28.2.490-498.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hay A. J., Joklik W. K. Demonstration that the same strand of reovirus genome RNA is transcribed in vitro and in vivo. Virology. 1971 May;44(2):450–453. doi: 10.1016/0042-6822(71)90276-5. [DOI] [PubMed] [Google Scholar]
  13. Kozak M. How do eucaryotic ribosomes select initiation regions in messenger RNA? Cell. 1978 Dec;15(4):1109–1123. doi: 10.1016/0092-8674(78)90039-9. [DOI] [PubMed] [Google Scholar]
  14. Kozak M. Nucleotide sequences of 5'-terminal ribosome-protected initiation regions from two reovirus messages. Nature. 1977 Sep 29;269(5627):391–394. doi: 10.1038/269390a0. [DOI] [PubMed] [Google Scholar]
  15. Kozak M., Shatkin A. J. Characterization of ribosome-protected fragments from reovirus messenger RNA. J Biol Chem. 1976 Jul 25;251(14):4259–4266. [PubMed] [Google Scholar]
  16. Kozak M., Shatkin A. J. Characterization of translational initiation regions from eukaryotic messenger RNAs. Methods Enzymol. 1979;60:360–375. doi: 10.1016/s0076-6879(79)60034-4. [DOI] [PubMed] [Google Scholar]
  17. Kozak M., Shatkin A. J. Identification of features in 5' terminal fragments from reovirus mRNA which are important for ribosome binding. Cell. 1978 Jan;13(1):201–212. doi: 10.1016/0092-8674(78)90150-2. [DOI] [PubMed] [Google Scholar]
  18. Kozak M., Shatkin A. J. Sequences and properties of two ribosome binding sites from the small size class of reovirus messenger RNA. J Biol Chem. 1977 Oct 10;252(19):6895–6908. [PubMed] [Google Scholar]
  19. Lhoas P. Mating pairs of Saccharomyces cerevisiae infected with double stranded RNA viruses from Aspergillus niger. Nat New Biol. 1972 Mar 22;236(64):86–87. doi: 10.1038/newbio236086a0. [DOI] [PubMed] [Google Scholar]
  20. McCrae M. A., Joklik W. K. The nature of the polypeptide encoded by each of the 10 double-stranded RNA segments of reovirus type 3. Virology. 1978 Sep;89(2):578–593. doi: 10.1016/0042-6822(78)90199-x. [DOI] [PubMed] [Google Scholar]
  21. Miura K., Watanabe K., Sugiura M., Shatkin A. J. The 5'-terminal nucleotide sequences of the double-stranded RNA of human reovirus. Proc Natl Acad Sci U S A. 1974 Oct;71(10):3979–3983. doi: 10.1073/pnas.71.10.3979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mustoe T. A., Ramig R. F., Sharpe A. H., Fields B. N. Genetics of reovirus: identification of the ds RNA segments encoding the polypeptides of the mu and sigma size classes. Virology. 1978 Sep;89(2):594–604. doi: 10.1016/0042-6822(78)90200-3. [DOI] [PubMed] [Google Scholar]
  23. Muthukrishnan S., Shatkin A. J. Reovirus genome RNA segments: resistance to S-1 nuclease. Virology. 1975 Mar;64(1):96–105. doi: 10.1016/0042-6822(75)90082-3. [DOI] [PubMed] [Google Scholar]
  24. Nonoyama M., Watanabe Y., Graham A. F. Defective virions of reovirus. J Virol. 1970 Aug;6(2):226–236. doi: 10.1128/jvi.6.2.226-236.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Peattie D. A. Direct chemical method for sequencing RNA. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1760–1764. doi: 10.1073/pnas.76.4.1760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pett D. M., Vanaman T. C., Joklik W. K. Studies on the amino and carboxyl terminal amino acid sequences of reovirus capsid polypeptides. Virology. 1973 Mar;52(1):174–186. doi: 10.1016/0042-6822(73)90407-8. [DOI] [PubMed] [Google Scholar]
  27. Pilly D., Niemeyer A., Schmidt M., Bargetzi J. P. Enzymes for RNA sequence analysis. Preparation and specificity of exoplasmodial ribonucleases I and II from Physarum polycephalum. J Biol Chem. 1978 Jan 25;253(2):437–445. [PubMed] [Google Scholar]
  28. Robertson J. S. 5' and 3' terminal nucleotide sequences of the RNA genome segments of influenza virus. Nucleic Acids Res. 1979 Aug 24;6(12):3745–3757. doi: 10.1093/nar/6.12.3745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Semancik J. S., Vidaver A. K., Van Etten J. L. Characterization of segmented double-helical RNA from bacteriophage phi6. J Mol Biol. 1973 Aug 25;78(4):617–625. doi: 10.1016/0022-2836(73)90283-0. [DOI] [PubMed] [Google Scholar]
  30. Shatkin A. J., Both G. W. Reovirus mRNA: transcription and translation. Cell. 1976 Mar;7(3):305–313. doi: 10.1016/0092-8674(76)90159-8. [DOI] [PubMed] [Google Scholar]
  31. Shatkin A. J. Capping of eucaryotic mRNAs. Cell. 1976 Dec;9(4 Pt 2):645–653. doi: 10.1016/0092-8674(76)90128-8. [DOI] [PubMed] [Google Scholar]
  32. Shatkin A. J., LaFiandra A. J. Transcription by infectious subviral particles of reovirus. J Virol. 1972 Oct;10(4):698–706. doi: 10.1128/jvi.10.4.698-706.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. 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]
  35. Silberklang M., Prochiantz A., Haenni A. L., Rajbhandary U. L. Studies on the sequence of the 3'-terminal region of turnip-yellow-mosaic-virus RNA. Eur J Biochem. 1977 Feb;72(3):465–478. doi: 10.1111/j.1432-1033.1977.tb11270.x. [DOI] [PubMed] [Google Scholar]
  36. Simoncsits A., Brownlee G. G., Brown R. S., Rubin J. R., Guilley H. New rapid gel sequencing method for RNA. Nature. 1977 Oct 27;269(5631):833–836. doi: 10.1038/269833a0. [DOI] [PubMed] [Google Scholar]
  37. Skehel J. J., Hay A. J. Nucleotide sequences at the 5' termini of influenza virus RNAs and their transcripts. Nucleic Acids Res. 1978 Apr;5(4):1207–1219. doi: 10.1093/nar/5.4.1207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sonenberg N., Morgan M. A., Merrick W. C., Shatkin A. J. A polypeptide in eukaryotic initiation factors that crosslinks specifically to the 5'-terminal cap in mRNA. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4843–4847. doi: 10.1073/pnas.75.10.4843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Stoltzfus C. M., Shatkin A. J., Banerjee A. K. Absence of polyadenylic acid from reovirus messenger ribonucleic acid. J Biol Chem. 1973 Dec 10;248(23):7993–7998. [PubMed] [Google Scholar]
  40. Weiner H. L., Ramig R. F., Mustoe T. A., Fields B. N. Identification of the gene coding for the hemagglutinin of reovirus. Virology. 1978 May 15;86(2):581–584. doi: 10.1016/0042-6822(78)90099-5. [DOI] [PubMed] [Google Scholar]

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