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
. 1969 Oct;64(2):626–633. doi: 10.1073/pnas.64.2.626

RNA SYNTHESIS IN CELLS INFECTED WITH HERPES SIMPLEX VIRUS, II. EVIDENCE THAT A CLASS OF VIRAL mRNA IS DERIVED FROM A HIGH MOLECULAR WEIGHT PRECURSOR SYNTHESIZED IN THE NUCLEUS*

Edward K Wagner 1,†, Bernard Roizman 1
PMCID: PMC223390  PMID: 4311377

Abstract

Viral RNA extracted from the cytoplasmic polyribosomes of human epidermoid carcinoma no. 2 cells infected with herpes simplex virus (mRNA) had a sedimentation coefficient between 10S and 20S while that from nuclei of infected cells varied in size from 10S to >80S. Estimates of the maximum molecular weight of viral RNA from its sedimentation coefficients suggest that at least 10 per cent of the viral genome is transcribed as a single molecule. The ratio of RNA of different sizes found in the nuclei of cells pulse labeled for 12 minutes was approximately the same as those found in cells labeled for longer intervals implying that either some classes of viral mRNA were made as small molecules or that the large viral RNA molecules were cleaved soon after synthesis. Cytoplasmic mRNA competed to a level of at least 80 per cent in viral DNA-RNA hybridization tests with >50S RNA extracted from nuclei of infected cells. This is consistent with the hypothesis that viral mRNA is produced by cleavage of a large precursor RNA molecule.

Full text

PDF
626

Selected References

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

  1. AURELIAN L., ROIZMAN B. ABORTIVE INFECTION OF CANINE CELLS BY HERPES SIMPLEX VIRUS. II. ALTERNATIVE SUPPRESSION OF SYNTHESIS OF INTERFERON AND VIRAL CONSTITUENTS. J Mol Biol. 1965 Mar;11:539–548. doi: 10.1016/s0022-2836(65)80009-2. [DOI] [PubMed] [Google Scholar]
  2. Borun T. W., Scharff M. D., Robbins E. Preparation of mammalian polyribosomes with the detergent Nonidet P-40. Biochim Biophys Acta. 1967 Nov 21;149(1):302–304. doi: 10.1016/0005-2787(67)90715-0. [DOI] [PubMed] [Google Scholar]
  3. GILBERT W. Polypeptide synthesis in Escherichia coli. II. The polypeptide chain and S-RNA. J Mol Biol. 1963 May;6:389–403. doi: 10.1016/s0022-2836(63)80051-0. [DOI] [PubMed] [Google Scholar]
  4. Gillespie D., Spiegelman S. A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J Mol Biol. 1965 Jul;12(3):829–842. doi: 10.1016/s0022-2836(65)80331-x. [DOI] [PubMed] [Google Scholar]
  5. Houssais J. F., Attardi G. High molecular weight nonribosomal-type nuclear RNA and cytoplasmic messenger RNA in HeLa cells. Proc Natl Acad Sci U S A. 1966 Aug;56(2):616–623. doi: 10.1073/pnas.56.2.616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Penman S., Smith I., Holtzman E. Ribosomal RNA synthesis and processing in a particulate site in the HeLa cell nucleus. Science. 1966 Nov 11;154(3750):786–789. doi: 10.1126/science.154.3750.786. [DOI] [PubMed] [Google Scholar]
  7. Penman S., Vesco C., Penman M. Localization and kinetics of formation of nuclear heterodisperse RNA, cytoplasmic heterodisperse RNA and polyribosome-associated messenger RNA in HeLa cells. J Mol Biol. 1968 May 28;34(1):49–60. doi: 10.1016/0022-2836(68)90234-9. [DOI] [PubMed] [Google Scholar]
  8. ROIZMAN B., AURELIAN L. ABORTIVE INFECTION OF CANINE CELLS BY HERPES SIMPLEX VIRUS. I. CHARACTERIZATION OF VIRAL PROGENY FROM CO-OPERATIVE INFECTION WITH MUTANTS DIFFERING IN CAPACITY TO MULTIPLY IN CANINE CELLS. J Mol Biol. 1965 Mar;11:528–538. doi: 10.1016/s0022-2836(65)80008-0. [DOI] [PubMed] [Google Scholar]
  9. STUDIER F. W. SEDIMENTATION STUDIES OF THE SIZE AND SHAPE OF DNA. J Mol Biol. 1965 Feb;11:373–390. doi: 10.1016/s0022-2836(65)80064-x. [DOI] [PubMed] [Google Scholar]
  10. Scherrer K., Marcaud L., Zajdela F., London I. M., Gros F. Patterns of RNA metabolism in a differentiated cell: a rapidly labeled, unstable 60S RNA with messenger properties in duck erythroblasts. Proc Natl Acad Sci U S A. 1966 Nov;56(5):1571–1578. doi: 10.1073/pnas.56.5.1571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Soeiro R., Amos H. Arrested protein synthesis in polysomes of cultured chick embryo cells. Science. 1966 Nov 4;154(3749):662–665. doi: 10.1126/science.154.3749.662. [DOI] [PubMed] [Google Scholar]
  12. Stévenin J., Mandel P., Jacob M. Relationship between nuclear giant-size dRNA and microsomal dRNA of rat brain. Proc Natl Acad Sci U S A. 1969 Feb;62(2):490–497. doi: 10.1073/pnas.62.2.490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. WARNER J. R., KNOPF P. M., RICH A. A multiple ribosomal structure in protein synthesis. Proc Natl Acad Sci U S A. 1963 Jan 15;49:122–129. doi: 10.1073/pnas.49.1.122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Wagner E. K., Roizman B. Ribonucleic acid synthesis in cells infected with herpes simplex virus. I. Patterns of ribonucleic acid synthesis in productively infected cells. J Virol. 1969 Jul;4(1):36–46. doi: 10.1128/jvi.4.1.36-46.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Warner J. R., Soeiro R., Birnboim H. C., Girard M., Darnell J. E. Rapidly labeled HeLa cell nuclear RNA. I. Identification by zone sedimentation of a heterogeneous fraction separate from ribosomal precursor RNA. J Mol Biol. 1966 Aug;19(2):349–361. doi: 10.1016/s0022-2836(66)80009-8. [DOI] [PubMed] [Google Scholar]
  16. Weinberg R. A., Loening U., Willems M., Penman S. Acrylamide gel electrophoresis of HeLa cell nucleolar RNA. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1088–1095. doi: 10.1073/pnas.58.3.1088. [DOI] [PMC free article] [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