Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1975 Jan;15(1):161–166. doi: 10.1128/jvi.15.1.161-166.1975

Molecular weight of RNA subunits of Rous sarcoma virus determined by electron microscopy.

A B Jacobson, P A Bromley
PMCID: PMC354430  PMID: 163340

Abstract

Secondary cultures of chicken embryo fibroblasts were infected with the Schmidt Ruppin strain of Rous sarcoma virus (RSV). Five days after infection, the medium was replaced at 2-h intervals with phosphate-free Eagle medium containing 50 muCi of [32P]orthophosphate per ml. Virus was collected by centrifugation, and the RNA was extracted and denatured with dimethyl sulfoxide, and the 33S subunit RNA was isolated by sucrose gradient centrifugation. The molecular weight of the RSV subunit RNA was determined by length measurement in the electron microscope, by using bacteriophage MS2 RNA as a length marker. Molecules of between 2.5 and 3.3 mum in length made up over 50% of the subunit RNA preparation. In this paper, we define RSV RNA subunits as that RNA released from the 70S RNA complex by dimethyl sulfoxide treatment, which sediments as a peak at 33S. Assuming the molecular weight of MS2 RNA to be 1.2 times 10-6, we calculate the molecular weight of RSV subunit RNA to be 3.12 times 10-6 plus or minus 0.25 times 10-6.

Full text

PDF
161

Images in this article

Selected References

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

  1. Bader J. P., Steck T. L. Analysis of the ribonucleic acid of murine leukemia virus. J Virol. 1969 Oct;4(4):454–459. doi: 10.1128/jvi.4.4.454-459.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Duesberg P. H. On the structure of RNA tumor viruses. Curr Top Microbiol Immunol. 1970;51:78–104. [PubMed] [Google Scholar]
  3. Duesberg P. H. Physical properties of Rous Sarcoma Virus RNA. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1511–1518. doi: 10.1073/pnas.60.4.1511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Duesberg P. H., Vogt P. K. Gel electrophoresis of avian leukosis and sarcoma viral RNA in formamide: comparison with other viral and cellular RNA species. J Virol. 1973 Sep;12(3):594–599. doi: 10.1128/jvi.12.3.594-599.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Erikson R. L. Studies on the RNA from avian myeloblastosis virus. Virology. 1969 Jan;37(1):124–131. doi: 10.1016/0042-6822(69)90313-4. [DOI] [PubMed] [Google Scholar]
  6. Fiers W., Lepoutre L., Vandendriessche L. Studies on the bacteriophage MS2. I. Distribution of purine sequences in the viral RNA and in yeast RNA. J Mol Biol. 1965 Sep;13(2):432–450. doi: 10.1016/s0022-2836(65)80108-5. [DOI] [PubMed] [Google Scholar]
  7. GESTELAND R. F., BOEDTKER H. SOME PHYSICAL PROPERTIES OF BACTERIOPHAGE R17 AND ITS RIBONUCLEIC ACID. J Mol Biol. 1964 Apr;8:496–507. doi: 10.1016/s0022-2836(64)80007-3. [DOI] [PubMed] [Google Scholar]
  8. Granboulan N., Huppert J., Lacour F. Examen au microscope electronique du RNA du virus de la myeloblastose aviaire. J Mol Biol. 1966 Apr;16(2):571–575. doi: 10.1016/s0022-2836(66)80196-1. [DOI] [PubMed] [Google Scholar]
  9. Haase A. T., Garapin A. C., Faras A. J., Taylor J. M., Bishop J. M. A comparison of the high molecular weight RNAs of visna virus and Rous sarcoma virus. Virology. 1974 Jan;57(1):259–270. doi: 10.1016/0042-6822(74)90126-3. [DOI] [PubMed] [Google Scholar]
  10. Kakefuda T., Bader J. P. Electron microscopic observations on the ribonucleic acid of murine leukemia virus. J Virol. 1969 Oct;4(4):460–474. doi: 10.1128/jvi.4.4.460-474.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kolakofsky D., Bruschi A. Molecular weight determination of Sendai RNA by dimethyl sulfoxide gradient sedimentation. J Virol. 1973 May;11(5):615–620. doi: 10.1128/jvi.11.5.615-620.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lang D. Molecular weights of coliphages and coliphage DNA. 3. Contour length and molecular weight of DNA from bacteriophages T4, T5 and T7, and from bovine papilloma virus. J Mol Biol. 1970 Dec 28;54(3):557–565. doi: 10.1016/0022-2836(70)90126-9. [DOI] [PubMed] [Google Scholar]
  13. Montagnier L., Goldé A., Vigier P. A possible subunit structure of Rous sarcoma virus RNA. J Gen Virol. 1969 Apr;4(3):449–452. doi: 10.1099/0022-1317-4-3-449. [DOI] [PubMed] [Google Scholar]
  14. Parsons J. T., Coffin J. M., Haroz R. K., Bromley P. A., Weissmann C. Quantitative determination and location of newly synthesized virus-specific ribonucleic acid in chicken cells infected with Rous sarcoma virus. J Virol. 1973 May;11(5):761–774. doi: 10.1128/jvi.11.5.761-774.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Robinson W. S., Pitkanen A., Rubin H. The nucleic acid of the Bryan strain of Rous sarcoma virus: purification of the virus and isolation of the nucleic acid. Proc Natl Acad Sci U S A. 1965 Jul;54(1):137–144. doi: 10.1073/pnas.54.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sarkar N. H., Moore D. H. Electron microscopy of the nucleic acid of mouse mammary tumor virus. J Virol. 1970 Feb;5(2):230–236. doi: 10.1128/jvi.5.2.230-236.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Scheele C. M., Hanafusa H. Electrophoretic analysis of the RNA of avian tumor viruses. Virology. 1972 Dec;50(3):753–764. doi: 10.1016/0042-6822(72)90429-1. [DOI] [PubMed] [Google Scholar]
  18. Taylor J. M., Varmus H. E., Faras A. J., Levinson W. E., Bishop J. M. Letter: Evidence for non-repetitive subunits in the genome of Rous sarcoma virus. J Mol Biol. 1974 Mar 25;84(1):217–221. doi: 10.1016/0022-2836(74)90223-x. [DOI] [PubMed] [Google Scholar]
  19. Trávnícek M., Ríman J. Subunits of oncornavirus high-molecular-weight RNA. I. Stepwise conversion of 60 S AMV (avian myeloblastosis virus) RNA to subunits. Biochem Biophys Res Commun. 1973 Jul 2;53(1):217–223. doi: 10.1016/0006-291x(73)91422-8. [DOI] [PubMed] [Google Scholar]
  20. Weber G. H., Heine U., Cottler-Fox M., Beaudreau G. S. Visualization of single-stranded nucleic acid of RNA tumor virus with the electron microscope. Proc Natl Acad Sci U S A. 1974 May;71(5):1887–1890. doi: 10.1073/pnas.71.5.1887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Whalley J. M. Electron microscopy of feline leukaemia virus RNA. Bibl Haematol. 1973;39:125–132. doi: 10.1159/000427834. [DOI] [PubMed] [Google Scholar]

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

RESOURCES