Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1975 Jun;15(6):1467–1474. doi: 10.1128/jvi.15.6.1467-1474.1975

Messenger activity of RNA transcribed in vitro by DNA-RNA polymerase associated to vaccinia virus cores.

G Jaureguiberry, F Ben-Hamida, F Chapeville, G Beaud
PMCID: PMC354614  PMID: 1170340

Abstract

The coding properties of RNA transcribed in vitro by purified vaccinia cores have been investigated using Krebs ascites tumor cells, L cells, and reticulocyte lysates. Six to 10 proteins synthesized in vitro are separated on polyacrylamide gels by electrophoresis in the presence of sodium dodecyl sulfate. Their molecular weights vary from 10,000 to 44,000. The electrophoretic behavior of these proteins is similar to that of early proteins isolated from infected L cells. The tryptic peptide analysis of one of these proteins indicates similarity in amino acid sequences. These results show fidelity of both in vitro transcription and molecular weight above 44,000 are synthesized in vitro does not seem due to a competition between 12S mRNA synthesized in excess and RNA of a higher sedimentation coefficient present in a lower amount.

Full text

PDF
1467

Images in this article

1469Image
on p.1469
1470Image
on p.1470
1472Image
on p.1472
1473Image
on p.1473

Selected References

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

  1. BECKER Y., JOKLIK W. K. MESSENGER RNA IN CELLS INFECTED WITH VACCINIA VIRUS. Proc Natl Acad Sci U S A. 1964 Apr;51:577–585. doi: 10.1073/pnas.51.4.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beaud G., Kirn A., Gros F. In vitro protein synthesis directed by RNA transcribed from vaccinia DNA. Biochem Biophys Res Commun. 1972 Dec 18;49(6):1459–1466. doi: 10.1016/0006-291x(72)90503-7. [DOI] [PubMed] [Google Scholar]
  3. Crawford L. V., Gesteland R. F. Synthesis of polyoma proteins in vitro. J Mol Biol. 1973 Mar 15;74(4):627–634. doi: 10.1016/0022-2836(73)90053-3. [DOI] [PubMed] [Google Scholar]
  4. Esteban M., Metz D. H. Early virus protein synthesis in vaccinia virus-infected cells. J Gen Virol. 1973 May;19(2):201–206. doi: 10.1099/0022-1317-19-2-201. [DOI] [PubMed] [Google Scholar]
  5. Fournier F., Tovell D. R., Esteban M., Metz D. H., Ball L. A., Kerr I. M. The translation of vaccinia virus messenger RNA in animal cell-free systems. FEBS Lett. 1973 Mar 15;30(3):268–272. doi: 10.1016/0014-5793(73)80667-2. [DOI] [PubMed] [Google Scholar]
  6. Graziadei W. D., 3rd, Roy D., Konigsberg W., Lengyel P. Translation of reovirus messenger ribonucleic acids synthesized in vitro into reovirus proteins in a mouse L cell extract. Arch Biochem Biophys. 1973 Sep;158(1):266–275. doi: 10.1016/0003-9861(73)90621-8. [DOI] [PubMed] [Google Scholar]
  7. Griffith I. P. Immediate visualization of proteins in dodecyl sulfate-polyacrylamide gels by prestaining with Remazol dyes. Anal Biochem. 1972 Apr;46(2):402–412. doi: 10.1016/0003-2697(72)90313-2. [DOI] [PubMed] [Google Scholar]
  8. Housman D., Jacobs-Lorena M., Rajbhandary U. L., Lodish H. F. Initiation of haemoglobin synthesis by methionyl-tRNA. Nature. 1970 Aug 29;227(5261):913–918. doi: 10.1038/227913a0. [DOI] [PubMed] [Google Scholar]
  9. JOKLIK W. K. THE INTRACELLULAR UNCOATING OF POXVIRUS DNA. II. THE MOLECULAR BASIS OF THE UNCOATING PROCESS. J Mol Biol. 1964 Feb;8:277–288. doi: 10.1016/s0022-2836(64)80137-6. [DOI] [PubMed] [Google Scholar]
  10. JOKLIK W. K. The purification fo four strains of poxvirus. Virology. 1962 Sep;18:9–18. doi: 10.1016/0042-6822(62)90172-1. [DOI] [PubMed] [Google Scholar]
  11. Joklik W. K. The poxviruses. Annu Rev Microbiol. 1968;22:359–390. doi: 10.1146/annurev.mi.22.100168.002043. [DOI] [PubMed] [Google Scholar]
  12. KIRN A., BRAUNWALD J., JUNG C., KLEIN M. INFLUENCE DE LA TEMP'ERATURE SUR LE D'EVELOPPEMENT DU VIRUS VACCINAL EN CELLULES KB. ETUDE PR'ELIMINAIRE. Pathol Microbiol (Basel) 1963;26:542–552. [PubMed] [Google Scholar]
  13. Kates J. R., McAuslan B. R. Messenger RNA synthesis by a "coated" viral genome. Proc Natl Acad Sci U S A. 1967 Feb;57(2):314–320. doi: 10.1073/pnas.57.2.314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kates J. R., McAuslan B. R. Poxvirus DNA-dependent RNA polymerase. Proc Natl Acad Sci U S A. 1967 Jul;58(1):134–141. doi: 10.1073/pnas.58.1.134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kates J., Beeson J. Ribonucleic acid synthesis in vaccinia virus. I. The mechanism of synthesis and release of RNA in vaccinia cores. J Mol Biol. 1970 May 28;50(1):1–18. doi: 10.1016/0022-2836(70)90100-2. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. MCAUSLAN B. R. THE INDUCTION AND REPRESSION OF THYMIDINE KINASE IN THE POXVIRUS-INFECTED HELA CELL. Virology. 1963 Nov;21:383–389. doi: 10.1016/0042-6822(63)90199-5. [DOI] [PubMed] [Google Scholar]
  18. Mathews M., Korner A. Mammalian cell-free protein synthesis directed by viral ribonucleic acid. Eur J Biochem. 1970 Dec;17(2):328–338. doi: 10.1111/j.1432-1033.1970.tb01170.x. [DOI] [PubMed] [Google Scholar]
  19. McDowell M. J., Joklik W. K., Villa-Komaroff L., Lodish H. F. Translation of reovirus messenger RNAs synthetesized in vitro into reovirus polypeptides by several mammalian cell-free extracts. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2649–2653. doi: 10.1073/pnas.69.9.2649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Metz D. H., Esteban M. Interferon inhibits viral protein synthesis in L cells infected with vaccinia virus. Nature. 1972 Aug 18;238(5364):385–388. doi: 10.1038/238385a0. [DOI] [PubMed] [Google Scholar]
  21. Moss B. Inhibition of HeLa cell protein synthesis by the vaccinia virion. J Virol. 1968 Oct;2(10):1028–1037. doi: 10.1128/jvi.2.10.1028-1037.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Moss B., Rosenblum E. N., Garon C. F. Glycoprotein synthesis in cells infected with vaccinia virus. 3. Purification and biosynthesis of the virion glycoprotein. Virology. 1973 Sep;55(1):143–156. doi: 10.1016/s0042-6822(73)81016-5. [DOI] [PubMed] [Google Scholar]
  23. Moss B., Salzman N. P. Sequential protein synthesis following vaccinia virus infection. J Virol. 1968 Oct;2(10):1016–1027. doi: 10.1128/jvi.2.10.1016-1027.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mss B., Filler R. Irreversible effects of cycloheximide during the early period of vaccinia virus replicaon. J Virol. 1970 Feb;5(2):99–108. doi: 10.1128/jvi.5.2.99-108.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Munyon W., Paoletti E., Grace J. T., Jr RNA polymerase activity in purified infectious vaccinia virus. Proc Natl Acad Sci U S A. 1967 Dec;58(6):2280–2287. doi: 10.1073/pnas.58.6.2280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Oda K. I., Joklik W. K. Hybridization and sedimentation studies on "early" and "late" vaccinia messenger RNA. J Mol Biol. 1967 Aug 14;27(3):395–419. doi: 10.1016/0022-2836(67)90047-2. [DOI] [PubMed] [Google Scholar]
  27. SHATKIN A. J. ACTINOMYCIN D AND VACCINIA VIRUS INFECTION OF HELA CELLS. Nature. 1963 Jul 27;199:357–358. doi: 10.1038/199357a0. [DOI] [PubMed] [Google Scholar]
  28. Salzman N. P., Sebring E. D. Sequential formation of vaccinia virus proteins and viral deoxyribonucleic acid replication. J Virol. 1967 Feb;1(1):16–23. doi: 10.1128/jvi.1.1.16-23.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sarov I., Joklik W. K. Characterization of intermediates in the uncoating of vaccinia virus DNA. Virology. 1972 Nov;50(2):593–602. doi: 10.1016/0042-6822(72)90410-2. [DOI] [PubMed] [Google Scholar]
  30. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  31. Woodson B. Recent progress in poxvirus research. Bacteriol Rev. 1968 Jun;32(2):127–137. doi: 10.1128/br.32.2.127-137.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES