1887

Journal of General Virology header logo

Volume 59, Issue 2

Synthesis of Prematurely Terminated Late Transcripts of Polyoma Virus DNA is Resistant to Inhibition by 5,6-Dichloro-1-β--ribofuranosylbenzimidazole Free

Abstract

SUMMARY

Exposure of mouse kidney cells to 5,6-dichloro-1-β--ribofuranosylbenzimidazole (DRB) (75 to 300 µ) during the late phase of infection by polyoma virus resulted in nearly complete (90 to 98%) inhibition of virus RNA synthesis. Sedimentation analysis revealed that, although the synthesis of high mol. wt. (> 10S) virus RNA was inhibited in a manner parallel to that of total virus RNA, the synthesis of small (3S to 7S) virus RNA was inhibited by only 40 to 50% in the presence of DRB. As a result, virus RNA synthesized in the presence of DRB contained a peak at 3S to 7S in addition to residual high mol. wt. virus RNA. Small virus RNA from either untreated or DRB-treated cells contained three- to sixfold higher levels of transcripts from the DNA fragment which lies between the HI and I sites (58.0 to 72.2 map units) than from DNA fragments covering the rest of the virus genome. Furthermore, 80% of the small RNA which hybridized to this fragment was complementary to the L strand of virus DNA. These results suggest that L strand transcripts are initiated within the I-HI DNA fragment and that a portion of these transcripts is prematurely terminated within several hundred nucleotides of the site(s) of initiation. DRB had little effect on the synthesis of these prematurely terminated RNAs.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): DNA tumour virus , inhibition , synthesis and transcription
© Journal of General Virology 1982
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-59-2-367
1982-04-01
2024-05-02
Loading full text...

Full text loading...

/deliver/fulltext/jgv/59/2/JV0590020367.html?itemId=/content/journal/jgv/10.1099/0022-1317-59-2-367&mimeType=html&fmt=ahah

References

  1. Acheson N. H. 1978; Polyoma virus giant RNAs contain tandem repeats of the nucleotide sequence of the entire viral genome. Proceedings of the National Academy of Sciences of the United States of America 75:4745–4748
     [Google Scholar]
  2. Acheson N. H. 1981; Efficiency of processing of viral RNA during the early and late phases of productive infection by polyoma virus. Journal of Virology 37:628–635
     [Google Scholar]
  3. Acheson N. H., Mieville F. 1978; Extent of transcription of the E strand of polyoma virus DNA during the early phase of productive infection. Journal of Virology 28:885–894
     [Google Scholar]
  4. Acheson N. H., Buetti E., Scherrer K., Weil R. 1971; Transcription of the polyoma virus genome: synthesis and cleavage of giant late polyoma-specific RNA. Proceedings of the National Academy of Sciences of the United States of America 68:2231–2235
     [Google Scholar]
  5. Bar-shavit R., Laub O., Aloni Y. 1978; The frequencies of transcription from the E and L strands of polyoma DNA. Journal of General Virology 39:357–360
     [Google Scholar]
  6. Beard P., Acheson N. H., Maxwell I. H. 1976; Strand-specific transcription of polyoma virus DNA early in productive infection and in transformed cells. Journal of Virology 17:20–26
     [Google Scholar]
  7. Contreras R., Fiers W. 1981; Initiation of transcription by RNA polymerase II in permeable, SV40-infected or noninfected, CV-1 cells: evidence for multiple promoters of SV40 late transcription. Nucleic Acids Research 9:215–236
     [Google Scholar]
  8. Dreyer C., Hausen P. 1978; On the activity of RNA polymerase B in lysates from Ehrlich ascites cells. European Journal of Biochemistry 86:241–253
     [Google Scholar]
  9. Egyhazi E. 1974; A tentative initiation inhibitor of chromosomal heterogeneous RNA synthesis. Journal of Molecular Biology 84:173–183
     [Google Scholar]
  10. Flavell A. J., Kamen R. 1977; Strand-specific transcription of polyoma virus DNA late during productive infection. Journal of Molecular Biology 115:237–242
     [Google Scholar]
  11. Flavell A. J., Cowie A., Legon S., Kamen R. 1979; Multiple 5′ terminal cap structures in late polyoma virus RNA. Cell 16:357–371
     [Google Scholar]
  12. Flavell A. J., Cowie A., Arrand J. R., Kamen R. I. 1980; Localization of three major capped 5′ ends of polyoma virus late mRNAs within a single tetranucleotide sequence in the viral genome. Journal of Virology 33:902–908
     [Google Scholar]
  13. Fraser N. W., Sehgal P. B., Darnell J. E. 1978; DRB-induced premature termination of late adenovirus transcription. Nature, London 272:590–593
     [Google Scholar]
  14. Fraser N. W., Sehgal P. B., darnell J. E. 1979; Multiple discrete sites for premature RNA chain termination late in adenovirus-2 infection: enhancement by 5, 6-dichloro-l-ß-Dribofuranosylbenzimidazole. Proceedings of the National Academy of Sciences of the United States of America 76:2571–2575
     [Google Scholar]
  15. Gidoni D., Kahana C., Canaani D., Groner Y. 1981; Specific in vitro initiation of transcription of simian virus 40 early and late genes occurs at the various cap nucleotides including cytidine. Proceedings of the National Academy of Sciences of the United States of America 78:2174–2178
     [Google Scholar]
  16. Granick D. 1975; Nucleolar necklaces in chick embryo fibroblast cells. 1. Formation of necklaces by dichlororibobenzimidazole and other adenosine analogues that decrease RNA synthesis and degrade preribosomes. Journal of Cell Biology 65:389–417
     [Google Scholar]
  17. Greenberg H., Penman S. 1966; Methylation and processing of ribosomal RNA in HeLa cells. Journal of Molecular Biology 21:527–535
     [Google Scholar]
  18. Handa H., Kaufman R. J., Manley J., Gefter M., Sharp P. 1981; Transcription of simian virus 40 DNA in a HeLa whole cell extract. Journal of Biological Chemistry 256:478–482
     [Google Scholar]
  19. Kamen R., Lindstrom H., Shure H., Old R. W. 1974; Virus-specific RNA in cells productively infected or transformed by polyoma virus. Cold Spring Harbor Symposia on Quantitative Biology 39:187–198
     [Google Scholar]
  20. Laub O., Jakobovits E. B., Aloni Y. 1980; 5, 6-dichloro-l-ß-d-ribofuranosylbenzimidazole enhances premature termination of late transcription of simian virus 40 DNA. Proceedings of the National Academy of Sciences of the United States of America 77:3297–3301
     [Google Scholar]
  21. Mcmaster G. K., Beard P., Engers H. D., Hirt B. 1981; Characterization of an immunosuppressive parvovirus related to the minute virus of mice. Journal of Virology 38:317–326
     [Google Scholar]
  22. Penman S. 1966; RNA metabolism in the HeLa cell nucleus. Journal of Molecular Biology 17:117–130
     [Google Scholar]
  23. Schibler U., Perry R. P. 1976; Characterization of the 5′ termini of hnRNA in mouse L cells: implications for processing and cap formation. Cell 9:121–130
     [Google Scholar]
  24. Sehgal P. B., Darnell J. E., Tamm I. 1976; The inhibition by DRB (5, 6-dichloro-1-ß-d-ribo-furanosylbenzimidazole) of hnRNA and mRNA production in HeLa cells. Cell 9:473–480
     [Google Scholar]
  25. Sehgal P. B., Fraser N. W., Darnell J. E. 1979; Early Ad-2 transcription units: only promoter-proximal RNA continues to be made in the presence of DRB. Virology 94:185–191
     [Google Scholar]
  26. Shmookler R. J., Buss J., Green M. H. 1974; Properties of the polyoma virus transcription complex obtained from mouse nuclei. Virology 57:122–127
     [Google Scholar]
  27. Spencer E., Loring J., Hurwitz J., Monroy G. 1978; unzymatic conversion of 5′-phosphate-terminated RNA to 5′-di- and triphosphate-terminated RNA. Proceedings of the National Academy of Sciences of the United States of America 75:4793–4797
     [Google Scholar]
  28. Tamm I. 1977; Definition of subclasses of nucleoplasmic RNA. Proceedings of the National Academy of Sciences of the United States of America 74:5011–5015
     [Google Scholar]
  29. Tamm I., Kikuchi T. 1979; Early termination of heterogeneous nuclear RNA transcripts in mammalian cells: accentuation by 5, 6-dichloro-l-ß-d-ribofuranosylbenzimidazole. Proceedings of the National Academy of Sciences of the United States of America 76:5750–5754
     [Google Scholar]
  30. Tamm L., Hand R., Caliguiri L. A. 1976; Action of dichlorobenzimidazole riboside on RNA synthesis in L-929 and HeLa cells. Journal of Cell Biology 69:229–240
     [Google Scholar]
  31. Tamm I., Kikuchi T., Darnell J. E., Salditt-Georgieff M. 1980; Short capped hnRNA precursor chains in HeLa cells: continued synthesis in the presence of 5, 6-dichloro-l-ß-d-ribofuranosylbenzimidazole. Biochemistry 19:2743–2748
     [Google Scholar]
  32. Treisman R. H. 1980; Characterization of polyoma late mRNA leader sequences by molecular cloning and DNA sequence analysis. Nucleic Acids Research 8:4867–4888
     [Google Scholar]
  33. Treisman R., Kamen R. 1981; Structure of polyoma virus late nuclear RNA. Journal of Molecular Biology 148:273–301
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-59-2-367
Loading
Synthesis of Prematurely Terminated Late Transcripts of Polyoma Virus DNA is Resistant to Inhibition by 5,6-Dichloro-1-β-d-ribofuranosylbenzimidazole
J. Gen. Virol. 59, 367 (1982); https://doi.org/10.1099/0022-1317-59-2-367
/content/journal/jgv/10.1099/0022-1317-59-2-367
/content/journal/jgv/10.1099/0022-1317-59-2-367
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error