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
. 1970 May;66(1):140–145. doi: 10.1073/pnas.66.1.140

Identity of the 5′-Terminal RNA Nucleotide Sequence of the Satellite Tobacco Necrosis Virus and Its Helper Virus: Possible Role of the 5′-Terminus in the Recognition by Virus-Specific RNA Replicase*

J A Lesnaw 1, M E Reichmann 1
PMCID: PMC286099  PMID: 5273892

Abstract

A pancreatic ribonuclease digest of 14C-labeled tobacco necrosis virus RNA was fractionated according to charge by column chromatography. Individual fractions were dephosphorylated with alkaline phosphatase and rechromatographed. The fraction, originally containing oligonucleotides with seven negative charges, separated into two components corresponding to five (-5) and two negative charges (-2). The -5 fraction was derived from the internal oligonucleotides while the -2 fraction must have originated from a 5′-pyrophosphorylated terminal trinucleotide. The sequence of this terminal trinucleotide was determined by column chromatography on DEAE-cellulose in a triethyl ammonium carbonate gradient, using the appropriate markers. The radioactivity chromatographed with a (ApGp)U marker. The order of the Ap and Gp was determined after ribonuclease T1 and alkaline phosphatase digestion. The radioactivity in the product chromatographed with an ApG marker. The 5′-terminus of tobacco necrosis virus RNA was therefore determined as ppApGpUp..., which is identical to the terminus of the RNA of its satellite virus as previously determined (J. Mol. Biol., 38, 59 (1968); Science, 160, 1452 (1968)). The 5′ pyrophosphate in both viruses was probably formed by an in vivo enzymatic removal of a γ-phosphate from a triphosphate, and its presence in both viruses suggested a common site of synthesis. The identity of the 5′-terminal sequences is considered not to be fortuitous and is discussed from the standpoint of their role as a recognition site for the virus-specific RNA replicase.

Full text

PDF
140

Selected References

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

  1. Adams J. M., Jeppesen P. G., Sanger F., Barrell B. G. Nucleotide sequence from the coat protein cistron of R17 bacteriophage RNA. Nature. 1969 Sep 6;223(5210):1009–1014. doi: 10.1038/2231009a0. [DOI] [PubMed] [Google Scholar]
  2. Bishop D. H., Mills D. R., Spiegelman S. The sequence at the 5' terminus of a self-replicating variant of viral Q-beta ribonucleic acid. Biochemistry. 1968 Oct;7(10):3744–3753. doi: 10.1021/bi00850a056. [DOI] [PubMed] [Google Scholar]
  3. Dahlberg J. E. Terminal sequences of bacteriophage RNAs. Nature. 1968 Nov 9;220(5167):548–552. doi: 10.1038/220548a0. [DOI] [PubMed] [Google Scholar]
  4. De Wachter R., Fiers W. Sequences at the 5'-terminus of bacteriophage Q-beta-RNA. Nature. 1969 Jan 18;221(5177):233–235. doi: 10.1038/221233a0. [DOI] [PubMed] [Google Scholar]
  5. Feix G., Pollet R., Weissmann C. Replication of viral RNA, XVI. Enzymatic synthesis of infectious virual RNA with noninfectious Q-beta minus strands as template. Proc Natl Acad Sci U S A. 1968 Jan;59(1):145–152. doi: 10.1073/pnas.59.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Glitz D. G. The nucleotide sequence at the 3'-linked end of bacteriophage MS2 ribonucleic acid. Biochemistry. 1968 Mar;7(3):927–932. doi: 10.1021/bi00843a007. [DOI] [PubMed] [Google Scholar]
  7. Haruna I., Spiegelman S. Recognition of size and sequence by an RNA replicase. Proc Natl Acad Sci U S A. 1965 Oct;54(4):1189–1193. doi: 10.1073/pnas.54.4.1189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lesnaw J. A., Reichmann M. E. The structure of tobacco necrosis virus. I. The protein subunit and the nature of the nucleic acid. Virology. 1969 Dec;39(4):729–737. doi: 10.1016/0042-6822(69)90010-5. [DOI] [PubMed] [Google Scholar]
  9. Lesnaw J. A., Reichmann M. E. The structure of tobacco necrosis virus. II. Terminal amino acid residues of the protein subunit. Virology. 1969 Dec;39(4):738–745. doi: 10.1016/0042-6822(69)90011-7. [DOI] [PubMed] [Google Scholar]
  10. Mills D. R., Peterson R. L., Spiegelman S. An extracellular Darwinian experiment with a self-duplicating nucleic acid molecule. Proc Natl Acad Sci U S A. 1967 Jul;58(1):217–224. doi: 10.1073/pnas.58.1.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. REICHMANN M. E. THE SATELLITE TOBACCO NECROSIS VIRUS: A SINGLE PROTEIN AND ITS GENETIC CODE. Proc Natl Acad Sci U S A. 1964 Oct;52:1009–1017. doi: 10.1073/pnas.52.4.1009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Roblin R. The 5'-terminus of bacteriophage R17 RNA: pppGp. J Mol Biol. 1968 Jan 14;31(1):51–61. doi: 10.1016/0022-2836(68)90053-3. [DOI] [PubMed] [Google Scholar]
  13. Roy D., Fraenkel-Conrat H., Lesnaw J., Reichmann M. E. The protein subunit of the satellite tobacco necrosis virus. Virology. 1969 Jun;38(2):368–369. doi: 10.1016/0042-6822(69)90384-5. [DOI] [PubMed] [Google Scholar]
  14. SUGIYAMA T., FRAENKEL-CONRAT H. The end-groups of tobacco mosaic virus RNA. II. Nature of the 3'-linked chain end in TMV and of both ends in four strains. Biochemistry. 1963 Mar-Apr;2:332–334. doi: 10.1021/bi00902a026. [DOI] [PubMed] [Google Scholar]
  15. Suzuki J., Haselkorn R. Studies on the 5'-terminus of turnip yellow mosaic virus RNA. J Mol Biol. 1968 Aug 28;36(1):47–56. doi: 10.1016/0022-2836(68)90218-0. [DOI] [PubMed] [Google Scholar]
  16. TOMLINSON R. V., TENER G. M. THE EFFECT OF UREA, FORMAMIDE, AND GLYCOLS ON THE SECONDARY BINDING FORCES IN THE ION-EXCHANGE CHROMATOGRAPHY OF POLYNUCLEOTIDES OF DEAE-CELLULOSE. Biochemistry. 1963 Jul-Aug;2:697–702. doi: 10.1021/bi00904a013. [DOI] [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. Wimmer E., Chang A. Y., Clark J. M., Jr, Reichmann M. E. Sequence studies of satellite tobacco necrosis virus RNA. Isolation and characterization of a 5'-terminal trinucleotide. J Mol Biol. 1968 Nov 28;38(1):59–73. doi: 10.1016/0022-2836(68)90128-9. [DOI] [PubMed] [Google Scholar]
  19. Wimmer E., Reichmann M. E. Pyrophosphate in the 5' terminal position of a viral ribonucleic acid. Science. 1968 Jun 28;160(3835):1452–1454. doi: 10.1126/science.160.3835.1452. [DOI] [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