Abstract
The rate of adenovirus RNA synthesis was compared in nuclei isolated from cells infected at 40.5°C in the presence of 1-β-d-arabinofuranosylcytosine with adenovirus 5 or an early temperature-sensitive mutant of adenovirus type 5, H5ts125 (ts125). In nuclei isolated at various times after infection, the maximum amount of virus RNA synthesis occurred at 6 h after infection, after which time virus RNA synthesis declined in nuclei from wild-type infections but remained high in nuclei from ts125 infections. At 12 h after infection, the amount of virus RNA synthesis was 8- to 11-fold higher in nuclei from ts125 infections than in nuclei from wild-type infections. However, the kinetics of virus RNA synthesis in nuclei isolated from both infections were similar. When a ts125-infected culture was shifted to 32°C for 3 h (12 to 15 h after infection) before nucleus isolation, the amount of virus RNA synthesis in the isolated nuclei was reduced to nearly wild-type levels. A pulse-chase experiment showed little difference in degradation rates of virus RNA in isolated nuclei from wild-type and ts125 infections. Hybridization of RNA synthesized in vitro to restriction fragments of adenovirus type 5 DNA was consistent with early virus RNA. These results support the idea that the 72,000-dalton DNA-binding protein encoded by the mutant gene in ts125 can regulate early adenovirus gene expression by inhibiting initiation of transcription of the adenovirus genome.
Full text
PDF







Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Alwine J. C., Reed S. I., Stark G. R. Characterization of the autoregulation of simian virus 40 gene A. J Virol. 1977 Oct;24(1):22–27. doi: 10.1128/jvi.24.1.22-27.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berget S. M., Flint S. J., Williams J. F., Sharp P. A. Adenovirus transcription. IV. Synthesis of viral-specific RNA in human cells infected with temperature-sensitive mutants of adenovirus 5. J Virol. 1976 Sep;19(3):879–889. doi: 10.1128/jvi.19.3.879-889.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berk A. J., Sharp P. A. Ultraviolet mapping of the adenovirus 2 early promoters. Cell. 1977 Sep;12(1):45–55. doi: 10.1016/0092-8674(77)90184-2. [DOI] [PubMed] [Google Scholar]
- Carter T. H., Blanton R. A. Autoregulation of adenovirus type 5 early gene expression II. Effect of temperature-sensitive early mutations on virus RNA accumulation. J Virol. 1978 Nov;28(2):450–456. doi: 10.1128/jvi.28.2.450-456.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carter T. H., Blanton R. A. Possible role of the 72,000 dalton DNA-binding protein in regulation of adenovirus type 5 early gene expression. J Virol. 1978 Feb;25(2):664–674. doi: 10.1128/jvi.25.2.664-674.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carter T. H., Ginsberg H. S. Viral transcription in KB cells infected by temperature-sensitive "early" mutants of adenovirus type 5. J Virol. 1976 Apr;18(1):156–166. doi: 10.1128/jvi.18.1.156-166.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Craig E. A., Raskas H. J. Nuclear transcripts larger than the cytoplasmic mRNAs are specified by segments of the adenovirus genome coding for early functions. Cell. 1976 Jun;8(2):205–213. doi: 10.1016/0092-8674(76)90004-0. [DOI] [PubMed] [Google Scholar]
- Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
- Ensinger M. J., Ginsberg H. S. Selection and preliminary characterization of temperature-sensitive mutants of type 5 adenovirus. J Virol. 1972 Sep;10(3):328–339. doi: 10.1128/jvi.10.3.328-339.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Flint S. J., Berget S. M., Sharp P. A. Adenovirus transcription. III. Mapping of viral RNA sequences in cells productively infected by adenovirus type 5. Virology. 1976 Jul 15;72(2):443–455. doi: 10.1016/0042-6822(76)90173-2. [DOI] [PubMed] [Google Scholar]
- Gilead Z., Sugawara K., Shanmugam G., Green M. Synthesis of the adenovirus-coded DNA binding protein in infected cells. J Virol. 1976 May;18(2):454–460. doi: 10.1128/jvi.18.2.454-460.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ginsberg H. S., Ensinger M. J., Kauffman R. S., Mayer A. J., Lundholm U. Cell transformation: a study of regulation with types 5 and 12 adenovirus temperature-sensitive mutants. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):419–426. doi: 10.1101/sqb.1974.039.01.054. [DOI] [PubMed] [Google Scholar]
- Ginsberg H. S., Lundholm U., Linné T. Adenovirus DNA-binding protein in cells infected with wild-type 5 adenovirus and two DNA-minus, temperature-sensitive mutants, H5ts125 and H5ts149. J Virol. 1977 Jul;23(1):142–151. doi: 10.1128/jvi.23.1.142-151.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levine A. J., Van Der Vliet P. C., Rosenwirth B., Rabek J., Frenkel G., Ensinger M. Adenovirus-infected, cell-specific, DNA-binding proteins. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):559–566. doi: 10.1101/sqb.1974.039.01.069. [DOI] [PubMed] [Google Scholar]
- Lewis J. B., Atkins J. F., Baum P. R., Solem R., Gesteland R. F., Anderson C. W. Location and identification of the genes for adenovirus type 2 early polypeptides. Cell. 1976 Jan;7(1):141–151. doi: 10.1016/0092-8674(76)90264-6. [DOI] [PubMed] [Google Scholar]
- Price R., Penman S. Transcription of the adenovirus genome by an -amanitine-sensitive ribonucleic acid polymerase in HeLa cells. J Virol. 1972 Apr;9(4):621–626. doi: 10.1128/jvi.9.4.621-626.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reed S. I., Stark G. R., Alwine J. C. Autoregulation of simian virus 40 gene A by T antigen. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3083–3087. doi: 10.1073/pnas.73.9.3083. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SCHERRER K., DARNELL J. E. Sedimentation characteristics of rapidly labelled RNA from HeLa cells. Biochem Biophys Res Commun. 1962 Jun 4;7:486–490. doi: 10.1016/0006-291x(62)90341-8. [DOI] [PubMed] [Google Scholar]
- Southern E. M. Long range periodicities in mouse satellite DNA. J Mol Biol. 1975 May 5;94(1):51–69. doi: 10.1016/0022-2836(75)90404-0. [DOI] [PubMed] [Google Scholar]
- Sussenbach J. S., Kuijk M. G. Studies on the mechanism of replication of adenovirus DNA. V. The location of termini of replication. Virology. 1977 Mar;77(1):149–157. doi: 10.1016/0042-6822(77)90414-7. [DOI] [PubMed] [Google Scholar]
- Tegtmeyer P. Altered patterns of protein synthesis in infection by SV40 mutants. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):9–15. doi: 10.1101/sqb.1974.039.01.004. [DOI] [PubMed] [Google Scholar]
- Tegtmeyer P., Schwartz M., Collins J. K., Rundell K. Regulation of tumor antigen synthesis by simain virus 40 gene A. J Virol. 1975 Jul;16(1):168–178. doi: 10.1128/jvi.16.1.168-178.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van Der Vliet P. C., Levine A. J., Ensinger M. J., Ginsberg H. S. Thermolabile DNA binding proteins from cells infected with a temperature-sensitive mutant of adenovrius defective in viral DNA synthesis. J Virol. 1975 Feb;15(2):348–354. doi: 10.1128/jvi.15.2.348-354.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vennström B., Pettersson U., Philipson L. Initiation of transcription in nuclei isolated from adenovirus infected cells. Nucleic Acids Res. 1978 Jan;5(1):205–219. doi: 10.1093/nar/5.1.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vennström B., Philipson L. Fidelity of adenovirus RNA transcription in isolated HeLa cell nuclei. J Virol. 1977 May;22(2):290–299. doi: 10.1128/jvi.22.2.290-299.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vliet P. C., Sussenbach J. S. An adenovirus type 5 gene function required for initiation of viral DNA replication. Virology. 1975 Oct;67(2):415–426. doi: 10.1016/0042-6822(75)90443-2. [DOI] [PubMed] [Google Scholar]
- Wallace R. D., Kates J. State of adenovirus 2 deoxyribonucleic acid in the nucleus and its mode of transcription: studies with isolated viral deoxyribonucleic acid-protein complexes and isolated nuclei. J Virol. 1972 Apr;9(4):627–635. doi: 10.1128/jvi.9.4.627-635.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weinmann R., Brendler T. G., Raskas H. J., Roeder R. G. Low molecular weight viral RNAs transcribed by RNA polymerase III during adenovirus 2 infection. Cell. 1976 Apr;7(4):557–566. doi: 10.1016/0092-8674(76)90206-3. [DOI] [PubMed] [Google Scholar]
- Weinmann R., Jaehning J. A., Raskas H. J., Roeder R. G. Viral RNA synthesis and levels of DNA-dependent RNA polymerases during replication of adenovirus 2. J Virol. 1975 Jan;17(1):114–126. doi: 10.1128/jvi.17.1.114-126.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van der Vliet P. C., Levine A. J. DNA-binding proteins specific for cells infected by adenovirus. Nat New Biol. 1973 Dec 12;246(154):170–174. doi: 10.1038/newbio246170a0. [DOI] [PubMed] [Google Scholar]