Abstract
An adenovirus (Ad) DNA replication complex extracted from infected HeLa nuclei could be purified free of the bulk of intracellular DNA polymerase activity by sedimetation in neutral sucrose gradients. However, the replication complex still retained some alpha and gamma DNA-polymerase activity. Since this complex is inhibited by 2', 3' dideoxythymidine-5'-triphosphate (ddTTP), an inhibitor of DNA polymerase gamma, a functional role for this enzyme in Ad DNA replication is suggested. Similar inhibition by ddTTP in intact Ad infected nuclei and comparable inhibition of Ad DNA synthesis in whole cells by dideoxythymidine (ddThy) are consistent with a role for DNA polymerase gamma. Uninfected HeLa nuclei or whole cells are not similarly inhibited by ddTTP or DDThy respectively. Such data does not rule out an additional functional role for other DNA polymerases, and recent experiments from this laboratory (1) suggest that DNA polymerase alpha is also involved in Ad DNA synthesis.
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Selected References
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- Abboud M. M., Sim W. J., Loeb L. A., Mildvan A. S. Apparent suicidal inactivation of DNA polymerase by adenosine 2',3'-riboepoxide 5'-triphosphate. J Biol Chem. 1978 May 25;253(10):3415–3421. [PubMed] [Google Scholar]
- Arens M., Yamashita T., Padmanabhan R., Tsuruo T., Green M. Adenovirus deoxyribonucleic acid replication. Characterization of the enzyme activities of a soluble replication system. J Biol Chem. 1977 Nov 25;252(22):7947–7954. [PubMed] [Google Scholar]
- Bolden A., Noy G. P., Weissbach A. DNA polymerase of mitochondria is a gamma-polymerase. J Biol Chem. 1977 May 25;252(10):3351–3356. [PubMed] [Google Scholar]
- Brison O., Kedinger C., Wilhelm J. Enzymatic properties of viral replication complexes isolated from adenovirus type 2-infected HeLa cell nuclei. J Virol. 1977 Nov;24(2):423–435. doi: 10.1128/jvi.24.2.423-435.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Byars N., Kidson C. DNA chain termination by 2',3'-dideoxythymidine in replicating mammalian cells. Biochemistry. 1975 Jul 15;14(14):3159–3164. doi: 10.1021/bi00685a019. [DOI] [PubMed] [Google Scholar]
- Dube D. K., Seal G., Loeb L. A. Differential heat sensitivity of mammalian DNA polymerases. Biochem Biophys Res Commun. 1976 May 23;76(2):483–487. doi: 10.1016/0006-291x(77)90750-1. [DOI] [PubMed] [Google Scholar]
- Edenberg H. J., Anderson S., DePamphilis M. L. Involvement of DNA polymerase alpha in simian virus 40 DNA replication. J Biol Chem. 1978 May 10;253(9):3273–3280. [PubMed] [Google Scholar]
- Frenkel G. D. Adenovirus DNA synthesis in vitro in an isolated complex. J Virol. 1978 Jan;25(1):459–463. doi: 10.1128/jvi.25.1.459-463.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horwitz M. S. Temperature-sensitive replication of H5ts125 adenovirus DNA in vitro. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4291–4295. doi: 10.1073/pnas.75.9.4291. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ito K., Arens M., Green M. Isolation of DNA polymerase gamma from an adenovirus 2 DNA replication complex. J Virol. 1975 Jun;15(6):1507–1510. doi: 10.1128/jvi.15.6.1507-1510.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaplan L. M., Kelinman R. E., Horwitz M. S. Replication of adenovirus type 2 DNA in vitro. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4425–4429. doi: 10.1073/pnas.74.10.4425. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lechner R. L., Kelly T. J., Jr The structure of replicating adenovirus 2 DNA molecules. Cell. 1977 Dec;12(4):1007–1020. doi: 10.1016/0092-8674(77)90165-9. [DOI] [PubMed] [Google Scholar]
- Maizel J. V., Jr, White D. O., Scharff M. D. The polypeptides of adenovirus. I. Evidence for multiple protein components in the virion and a comparison of types 2, 7A, and 12. Virology. 1968 Sep;36(1):115–125. doi: 10.1016/0042-6822(68)90121-9. [DOI] [PubMed] [Google Scholar]
- Matsukage A., Bohn E. W., Wilson S. H. Differential sensitivity of low molecular weight DNA polymerase to sulfhydryl-blocking reagents. Biochim Biophys Acta. 1975 Mar 21;383(3):338–343. doi: 10.1016/0005-2787(75)90062-3. [DOI] [PubMed] [Google Scholar]
- Matsukage A., Bohn E. W., Wilson S. H. On the DNA polymerase III of mouse myeloma: partial purification and characterization. Biochemistry. 1975 Mar 11;14(5):1006–1020. doi: 10.1021/bi00676a020. [DOI] [PubMed] [Google Scholar]
- Spadari S., Weissbach A. The interrelation between DNA synthesis and various DNA polymerase activities in synchronized HeLa cells. J Mol Biol. 1974 Jun 15;86(1):11–20. doi: 10.1016/s0022-2836(74)80003-3. [DOI] [PubMed] [Google Scholar]
- Tremblay G. Y., Daniels M. J., Schaechter M. Isolation of a cell membrane-DNA-nascent RNA complex from bacteria. J Mol Biol. 1969 Feb 28;40(1):65–76. doi: 10.1016/0022-2836(69)90296-4. [DOI] [PubMed] [Google Scholar]
- Waqar M. A., Evans M. J., Huberman J. A. Effect of 2',3'-dideoxythymidine-5'-triphosphate on HeLa cell in vitro DNA synthesis: evidence that DNA polymerase alpha is the only polymerase required for cellular DNA replication. Nucleic Acids Res. 1978 Jun;5(6):1933–1946. doi: 10.1093/nar/5.6.1933. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weissbach A. Eukaryotic DNA polymerases. Annu Rev Biochem. 1977;46:25–47. doi: 10.1146/annurev.bi.46.070177.000325. [DOI] [PubMed] [Google Scholar]
- Yamashita T., Arens M., Green M. Adenovirus deoxyribonucleic acid replication. Isolation of a soluble replication system and analysis of the in vitro DNA product. J Biol Chem. 1977 Nov 25;252(22):7940–7946. [PubMed] [Google Scholar]
- van der Vliet P. C., Kwant M. M. Role of DNA polymerase gamma in adenovirus DNA replication. Nature. 1978 Nov 30;276(5687):532–534. doi: 10.1038/276532a0. [DOI] [PubMed] [Google Scholar]