Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1983 Feb 25;11(4):1039–1058. doi: 10.1093/nar/11.4.1039

Polyoma-induced stimulation of nucleoplasmic transcription is paralleled by development of resistance against actinomycin D.

J M Matter, E W Khandjian, R Weil
PMCID: PMC325775  PMID: 6298729

Abstract

Polyoma virus induced in quiescent, Go-arrested mouse kidney cells a lytic infection. Synthesis of the polyoma T-antigens began 7-8 h after infection and was followed by a mitotic reaction of the host cell comprising stimulated synthesis and accumulation of cellular (mainly ribosomal) RNA and protein and duplication of the host cell chromatin (S-phase). In the present work we focused attention on nucleoplasmic transcription, i.e. synthesis of hnRNA, 5S RNA and tRNA. To inhibit selectively nucleolar transcription we used low concentrations of actinomycin D (act. D). Synthesis of 45S precursor- ribosomal RNA in mock- and polyoma-infected mouse kidney cells was completely blocked by 0.05 micrograms/ml act.D within 2 h. In mock-infected cells also nucleoplasmic transcription was rather sensitive against 0.05 micrograms/ml act.D. Polyoma- induced stimulation of nucleoplasmic transcription began around 12 h and was paralleled by the development of resistance against act.D. Resistance of nucleoplasmic transcription in virus-infected cells was thus similar to that observed by others in uninfected, proliferating mammalian cells. The possible biological implications of these results are discussed.

Full text

PDF
1039

Images in this article

1046Image
on p.1046
1053Image
on p.1053

Selected References

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

  1. Bernhardt D., Darnell J. E., Jr tRNA synthesis in HeLa cells: a precursor to tRNA and the effects of methionine starvation on tRNA synthesis. J Mol Biol. 1969 May 28;42(1):43–56. doi: 10.1016/0022-2836(69)90485-9. [DOI] [PubMed] [Google Scholar]
  2. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  3. Buetti E. Characterization of late polyoma mRNA. J Virol. 1974 Aug;14(2):249–260. doi: 10.1128/jvi.14.2.249-260.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DULBECCO R., HARTWELL L. H., VOGT M. INDUCTION OF CELLULAR DNA SYNTHESIS BY POLYOMA VIRUS. Proc Natl Acad Sci U S A. 1965 Feb;53:403–410. doi: 10.1073/pnas.53.2.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fogel M., Gilden R., Defendi V. Polyoma virus-induced "complement-fixing antigen" in tumors and infected cells and detected by immunofluorescence. Proc Soc Exp Biol Med. 1967 Apr;124(4):1047–1052. doi: 10.3181/00379727-124-31920. [DOI] [PubMed] [Google Scholar]
  6. GEORGIEV G. P., SAMARINA O. P., LERMAN M. I., SMIRNOV M. N., SEVERTZOV A. N. BIOSYNTHESIS OF MESSENGER AND RIBOSOMAL RIBONUCLEIC ACIDS IN THE NUCLEOLOCHROMOSOMAL APPARATUS OF ANIMAL CELLS. Nature. 1963 Dec 28;200:1291–1294. doi: 10.1038/2001291a0. [DOI] [PubMed] [Google Scholar]
  7. Hancock R., Weil R. Biochemical evidence for induction by polyoma virus of replication of the chromosomes of mouse kidney cells. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1144–1150. doi: 10.1073/pnas.63.4.1144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hodnett J. L., Busch H. Isolation and characterization of uridylic acid-rich 7 S ribonucleic acid of rat liver nuclei. J Biol Chem. 1968 Dec 25;243(24):6334–6342. [PubMed] [Google Scholar]
  9. Khandjian E. W., Matter J. M., Léonard N., Weil R. Simian virus 40 and polyoma virus stimulate overall cellular RNA and protein synthesis. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1476–1480. doi: 10.1073/pnas.77.3.1476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kára J., Weil R. Specific activation of the DNA-synthesizing apparatus in contact-inhibited mouse kidney cells by polyoma virus. Proc Natl Acad Sci U S A. 1967 Jan;57(1):63–70. doi: 10.1073/pnas.57.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lindell T. J. Evidence for an extranucleolar mechanism of actinomycin D action. Nature. 1976 Sep 23;263(5575):347–350. doi: 10.1038/263347a0. [DOI] [PubMed] [Google Scholar]
  12. MASTER R. W. POSSIBLE SYNTHESIS OF POLYRIBONUCLEOTIDES OF KNOWN BASE-TRIPLET SEQUENCES. Nature. 1965 Apr 3;206:93–93. doi: 10.1038/206093b0. [DOI] [PubMed] [Google Scholar]
  13. Matter J. M., Weil R. The relation between polyoma T-antigen and increased 5S RNA synthesis in cell-free extracts from polyoma-infected mouse kidney cell cultures. Nucleic Acids Res. 1982 Dec 11;10(23):7643–7655. doi: 10.1093/nar/10.23.7643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mirault M. E., Scherrer K. Isolation of preribosomes from HeLa cells and their characterization by electrophoresis on uniform and exponential-gradient-polyacrylamide gels. Eur J Biochem. 1971 Nov 11;23(2):372–386. doi: 10.1111/j.1432-1033.1971.tb01631.x. [DOI] [PubMed] [Google Scholar]
  15. Palmiter R. D., Schimke R. T. Regulation of protein synthesis in chick oviduct. 3. Mechanism of ovalbumin "superinduction" by actinomycin D. J Biol Chem. 1973 Mar 10;248(5):1502–1512. [PubMed] [Google Scholar]
  16. Peacock A. C., Dingman C. W. Molecular weight estimation and separation of ribonucleic acid by electrophoresis in agarose-acrylamide composite gels. Biochemistry. 1968 Feb;7(2):668–674. doi: 10.1021/bi00842a023. [DOI] [PubMed] [Google Scholar]
  17. Pederson T., Robbins E. Chromatin structure and the cell division cycle. Actinomycin binding in synchronized HeLa cells. J Cell Biol. 1972 Nov;55(2):322–327. doi: 10.1083/jcb.55.2.322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pene J. J., Knight E., Jr, Darnell J. E., Jr Characterization of a new low molecular weight RNA in HeLa cell ribosomes. J Mol Biol. 1968 May 14;33(3):609–623. doi: 10.1016/0022-2836(68)90309-4. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Perry R. P., Kelley D. E. Persistent synthesis of 5S RNA when production of 28S and 18S ribosomal RNA is inhibited by low doses of actinomycin D. J Cell Physiol. 1968 Dec;72(3):235–246. doi: 10.1002/jcp.1040720311. [DOI] [PubMed] [Google Scholar]
  21. Perry R. P., La Torre J., Kelley D. E., Greenberg J. R. On the lability of poly(A) sequences during extraction of messenger RNA from polyribosomes. Biochim Biophys Acta. 1972 Mar 14;262(2):220–226. doi: 10.1016/0005-2787(72)90236-5. [DOI] [PubMed] [Google Scholar]
  22. Pruitt S. C., Grainger R. M. A mosaicism in the higher order structure of Xenopus oocyte nucleolar chromatin prior to and during ribosomal gene transcription. Cell. 1981 Mar;23(3):711–720. doi: 10.1016/0092-8674(81)90434-7. [DOI] [PubMed] [Google Scholar]
  23. Pétursson G., Weil R. A study on the mechanism of polyoma-induced activation of the cellular DNA-synthesizing apparatus. Synchronization by FUdR of virus-induced DNA synthesis. Arch Gesamte Virusforsch. 1968;24(1):1–29. doi: 10.1007/BF01242898. [DOI] [PubMed] [Google Scholar]
  24. Pöckl E., Wintersberger E. Increased rate of RNA synthesis: early reaction of primary mouse kidney cells to infection with polyoma virus of simian virus 40. J Virol. 1980 Jul;35(1):8–19. doi: 10.1128/jvi.35.1.8-19.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Reijnders L., Sloof P., Sival J., Borst P. Gel electrophoresis of RNA under denaturing conditions. Biochim Biophys Acta. 1973 Oct 26;324(3):320–333. doi: 10.1016/0005-2787(73)90278-5. [DOI] [PubMed] [Google Scholar]
  26. Roberts W. K., Newman J. F. Use of low concentrations of actinomycin D in the study of RNA synthesis in Ehrlich ascites cells. J Mol Biol. 1966 Sep;20(1):63–73. doi: 10.1016/0022-2836(66)90117-3. [DOI] [PubMed] [Google Scholar]
  27. Rosenthal L. J. Isolation and characterization of poly(A)-containing polyoma "early" and "late" messenger RNAs. Nucleic Acids Res. 1976 Mar;3(3):661–676. doi: 10.1093/nar/3.3.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Salomon C., Türler H., Weil R. Polyoma-induced stimulation of cellular RNA synthesis is paralleled by changed expression of the viral genome. Nucleic Acids Res. 1977;4(5):1483–1503. doi: 10.1093/nar/4.5.1483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Scherrer K., Imaizumi-Scherrer M. T., Reynaud C. A., Therwath A. On pre-messenger RNA and transcriptions. A review. Mol Biol Rep. 1979 May 31;5(1-2):5–28. doi: 10.1007/BF00777484. [DOI] [PubMed] [Google Scholar]
  30. Schwyzer M., Weil R., Frank G., Zuber H. Amino acid sequence analysis of fragments generated by partial proteolysis from large simian virus 40 tumor antigen. J Biol Chem. 1980 Jun 25;255(12):5627–5634. [PubMed] [Google Scholar]
  31. Sobell H. M., Jain S. C., Sakore T. D., Nordman C. E. Stereochemistry of actinomycin--DNA binding. Nat New Biol. 1971 Jun 16;231(24):200–205. doi: 10.1038/newbio231200a0. [DOI] [PubMed] [Google Scholar]
  32. Türler H. The tumor antigens and the early functions of polyoma virus. Mol Cell Biochem. 1980 Sep 15;32(2):63–93. doi: 10.1007/BF00227801. [DOI] [PubMed] [Google Scholar]
  33. WINOCOUR E. Purification of polyoma virus. Virology. 1963 Feb;19:158–168. doi: 10.1016/0042-6822(63)90005-9. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Weil P. A., Segall J., Harris B., Ng S. Y., Roeder R. G. Faithful transcription of eukaryotic genes by RNA polymerase III in systems reconstituted with purified DNA templates. J Biol Chem. 1979 Jul 10;254(13):6163–6173. [PubMed] [Google Scholar]
  36. Weil R., Kára J. Polyoma "tumor antigen": an activator of chromosome replication? Proc Natl Acad Sci U S A. 1970 Oct;67(2):1011–1017. doi: 10.1073/pnas.67.2.1011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weil R., Michel M. R., Ruschmann G. K. Induction of cellular DNA synthesis by polyoma virus. Proc Natl Acad Sci U S A. 1965 Jun;53(6):1468–1475. doi: 10.1073/pnas.53.6.1468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weil R. Viral 'tumor antigens': A novel type of mammalian regulator protein. Biochim Biophys Acta. 1978 Nov 17;516(3):301–388. doi: 10.1016/0304-419x(78)90012-4. [DOI] [PubMed] [Google Scholar]
  39. Weinberg R. A. Nuclear RNA metabolism. Annu Rev Biochem. 1973;42:329–354. doi: 10.1146/annurev.bi.42.070173.001553. [DOI] [PubMed] [Google Scholar]
  40. Weinberg R. A., Penman S. Small molecular weight monodisperse nuclear RNA. J Mol Biol. 1968 Dec;38(3):289–304. doi: 10.1016/0022-2836(68)90387-2. [DOI] [PubMed] [Google Scholar]
  41. Wintersberger E., Wintersberger U. Induction of DNA polymerase in polyoma virus-infected mouse cells requires transcription and translation. J Virol. 1976 Aug;19(2):291–295. doi: 10.1128/jvi.19.2.291-295.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES