Abstract
Host cell cycle genes provide important functions to retroviruses and retroviruslike elements. To define some of these functions, the cell cycle dependence of transposition of the yeast retroviruslike element Ty3 was examined. Ty3 is unique among retroviruslike elements because of the specificity of its integration, which occurs upstream of genes transcribed by RNA polymerase III. A physical assay for Ty3 transposition which takes advantage of this position-specific integration was developed. The assay uses PCR to amplify a product of Ty3 integration into a target plasmid that carries a modified tRNA gene. By using the GAL1 upstream activating sequence to regulate expression of Ty3, transposition was detected within one generation of cell growth after Ty3 transcription was initiated. This physical assay was used to show that Ty3 did not transpose when yeast cells were arrested in G1 during treatment with the mating pheromone alpha-factor. The restriction of transposition was not due to changes in transcription of either Ty3 or tRNA genes or to aspects of the mating pheromone response unrelated to cell cycle arrest. The block of the Ty3 life cycle was reversed when cells were released from G1 arrest. Examination of Ty3 intermediates during G1 arrest indicated that Ty3 viruslike particles were present but that reverse transcription of the Ty3 genomic RNA into double-stranded DNA had not occurred. In G1, the Ty3 life cycle is blocked after particle assembly but before the completion of reverse transcription.
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.
- Adams B. G. Induction of galactokinase in Saccharomyces cerevisiae: kinetics of induction and glucose effects. J Bacteriol. 1972 Aug;111(2):308–315. doi: 10.1128/jb.111.2.308-315.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Alani E., Cao L., Kleckner N. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Genetics. 1987 Aug;116(4):541–545. doi: 10.1534/genetics.112.541.test. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bilanchone V. W., Claypool J. A., Kinsey P. T., Sandmeyer S. B. Positive and negative regulatory elements control expression of the yeast retrotransposon Ty3. Genetics. 1993 Jul;134(3):685–700. doi: 10.1093/genetics/134.3.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boeke J. D., LaCroute F., Fink G. R. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. doi: 10.1007/BF00330984. [DOI] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Bukrinsky M. I., Haggerty S., Dempsey M. P., Sharova N., Adzhubel A., Spitz L., Lewis P., Goldfarb D., Emerman M., Stevenson M. A nuclear localization signal within HIV-1 matrix protein that governs infection of non-dividing cells. Nature. 1993 Oct 14;365(6447):666–669. doi: 10.1038/365666a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bukrinsky M. I., Sharova N., Dempsey M. P., Stanwick T. L., Bukrinskaya A. G., Haggerty S., Stevenson M. Active nuclear import of human immunodeficiency virus type 1 preintegration complexes. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6580–6584. doi: 10.1073/pnas.89.14.6580. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chalker D. L., Sandmeyer S. B. Sites of RNA polymerase III transcription initiation and Ty3 integration at the U6 gene are positioned by the TATA box. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):4927–4931. doi: 10.1073/pnas.90.11.4927. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chalker D. L., Sandmeyer S. B. Transfer RNA genes are genomic targets for de Novo transposition of the yeast retrotransposon Ty3. Genetics. 1990 Dec;126(4):837–850. doi: 10.1093/genetics/126.4.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chalker D. L., Sandmeyer S. B. Ty3 integrates within the region of RNA polymerase III transcription initiation. Genes Dev. 1992 Jan;6(1):117–128. doi: 10.1101/gad.6.1.117. [DOI] [PubMed] [Google Scholar]
- Chang F., Herskowitz I. Identification of a gene necessary for cell cycle arrest by a negative growth factor of yeast: FAR1 is an inhibitor of a G1 cyclin, CLN2. Cell. 1990 Nov 30;63(5):999–1011. doi: 10.1016/0092-8674(90)90503-7. [DOI] [PubMed] [Google Scholar]
- Clark D. J., Bilanchone V. W., Haywood L. J., Dildine S. L., Sandmeyer S. B. A yeast sigma composite element, TY3, has properties of a retrotransposon. J Biol Chem. 1988 Jan 25;263(3):1413–1423. [PubMed] [Google Scholar]
- Coney L. R., Roeder G. S. Control of yeast gene expression by transposable elements: maximum expression requires a functional Ty activator sequence and a defective Ty promoter. Mol Cell Biol. 1988 Oct;8(10):4009–4017. doi: 10.1128/mcb.8.10.4009. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fritsch E. F., Temin H. M. Inhibition of viral DNA synthesis in stationary chicken embryo fibroblasts infected with avian retroviruses. J Virol. 1977 Nov;24(2):461–469. doi: 10.1128/jvi.24.2.461-469.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fritsch E., Temin H. M. Formation and structure of infectious DNA of spleen necrosis virus. J Virol. 1977 Jan;21(1):119–130. doi: 10.1128/jvi.21.1.119-130.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gartner S., Markovits P., Markovitz D. M., Kaplan M. H., Gallo R. C., Popovic M. The role of mononuclear phagocytes in HTLV-III/LAV infection. Science. 1986 Jul 11;233(4760):215–219. doi: 10.1126/science.3014648. [DOI] [PubMed] [Google Scholar]
- Hansen L. J., Chalker D. L., Orlinsky K. J., Sandmeyer S. B. Ty3 GAG3 and POL3 genes encode the components of intracellular particles. J Virol. 1992 Mar;66(3):1414–1424. doi: 10.1128/jvi.66.3.1414-1424.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hansen L. J., Chalker D. L., Sandmeyer S. B. Ty3, a yeast retrotransposon associated with tRNA genes, has homology to animal retroviruses. Mol Cell Biol. 1988 Dec;8(12):5245–5256. doi: 10.1128/mcb.8.12.5245. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hansen L. J., Sandmeyer S. B. Characterization of a transpositionally active Ty3 element and identification of the Ty3 integrase protein. J Virol. 1990 Jun;64(6):2599–2607. doi: 10.1128/jvi.64.6.2599-2607.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harel J., Rassart E., Jolicoeur P. Cell cycle dependence of synthesis of unintegrated viral DNA in mouse cells newly infected with murine leukemia virus. Virology. 1981 Apr 15;110(1):202–207. doi: 10.1016/0042-6822(81)90022-2. [DOI] [PubMed] [Google Scholar]
- Ho D. D., Rota T. R., Hirsch M. S. Infection of monocyte/macrophages by human T lymphotropic virus type III. J Clin Invest. 1986 May;77(5):1712–1715. doi: 10.1172/JCI112491. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hsu T. W., Taylor J. M. Effect of aphidicolin on avian sarcoma virus replication. J Virol. 1982 Nov;44(2):493–498. doi: 10.1128/jvi.44.2.493-498.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ihle J. N., Kenney F. T., Tennant R. W. Evidence for a stable intermediate in leukemia virus activation in AKR mouse embryo cells. J Virol. 1974 Sep;14(3):451–456. doi: 10.1128/jvi.14.3.451-456.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johnston M. A model fungal gene regulatory mechanism: the GAL genes of Saccharomyces cerevisiae. Microbiol Rev. 1987 Dec;51(4):458–476. doi: 10.1128/mr.51.4.458-476.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johnston M., Davis R. W. Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Aug;4(8):1440–1448. doi: 10.1128/mcb.4.8.1440. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Keil R. L., Roeder G. S. Cis-acting, recombination-stimulating activity in a fragment of the ribosomal DNA of S. cerevisiae. Cell. 1984 Dec;39(2 Pt 1):377–386. doi: 10.1016/0092-8674(84)90016-3. [DOI] [PubMed] [Google Scholar]
- Kirchner J., Sandmeyer S. B., Forrest D. B. Transposition of a Ty3 GAG3-POL3 fusion mutant is limited by availability of capsid protein. J Virol. 1992 Oct;66(10):6081–6092. doi: 10.1128/jvi.66.10.6081-6092.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kirchner J., Sandmeyer S. Proteolytic processing of Ty3 proteins is required for transposition. J Virol. 1993 Jan;67(1):19–28. doi: 10.1128/jvi.67.1.19-28.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kitagawa T., Aikawa T. Enzyme coupled immunoassay of insulin using a novel coupling reagent. J Biochem. 1976 Jan;79(1):233–236. doi: 10.1093/oxfordjournals.jbchem.a131053. [DOI] [PubMed] [Google Scholar]
- Lewis P., Hensel M., Emerman M. Human immunodeficiency virus infection of cells arrested in the cell cycle. EMBO J. 1992 Aug;11(8):3053–3058. doi: 10.1002/j.1460-2075.1992.tb05376.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liu F. T., Zinnecker M., Hamaoka T., Katz D. H. New procedures for preparation and isolation of conjugates of proteins and a synthetic copolymer of D-amino acids and immunochemical characterization of such conjugates. Biochemistry. 1979 Feb 20;18(4):690–693. doi: 10.1021/bi00571a022. [DOI] [PubMed] [Google Scholar]
- MacKay V. L., Welch S. K., Insley M. Y., Manney T. R., Holly J., Saari G. C., Parker M. L. The Saccharomyces cerevisiae BAR1 gene encodes an exported protein with homology to pepsin. Proc Natl Acad Sci U S A. 1988 Jan;85(1):55–59. doi: 10.1073/pnas.85.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McDougal J. S., Mawle A., Cort S. P., Nicholson J. K., Cross G. D., Scheppler-Campbell J. A., Hicks D., Sligh J. Cellular tropism of the human retrovirus HTLV-III/LAV. I. Role of T cell activation and expression of the T4 antigen. J Immunol. 1985 Nov;135(5):3151–3162. [PubMed] [Google Scholar]
- Mullis K. B., Faloona F. A. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. doi: 10.1016/0076-6879(87)55023-6. [DOI] [PubMed] [Google Scholar]
- Nasmyth K. Regulating the HO endonuclease in yeast. Curr Opin Genet Dev. 1993 Apr;3(2):286–294. doi: 10.1016/0959-437x(93)90036-o. [DOI] [PubMed] [Google Scholar]
- Parent S. A., Fenimore C. M., Bostian K. A. Vector systems for the expression, analysis and cloning of DNA sequences in S. cerevisiae. Yeast. 1985 Dec;1(2):83–138. doi: 10.1002/yea.320010202. [DOI] [PubMed] [Google Scholar]
- RUBIN H., TEMIN H. M. A radiological study of cell-virus interaction in the Rous sarcoma. Virology. 1959 Jan;7(1):75–91. doi: 10.1016/0042-6822(59)90178-3. [DOI] [PubMed] [Google Scholar]
- Richter A., Ozer H. L., DesGroseillers L., Jolicoeur P. An X-linked gene affecting mouse cell DNA synthesis also affects production of unintegrated linear and supercoiled DNA of murine leukemia virus. Mol Cell Biol. 1984 Jan;4(1):151–159. doi: 10.1128/mcb.4.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roe T., Reynolds T. C., Yu G., Brown P. O. Integration of murine leukemia virus DNA depends on mitosis. EMBO J. 1993 May;12(5):2099–2108. doi: 10.1002/j.1460-2075.1993.tb05858.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shah H. C., Carlson G. P. Alteration by phenobarbital and 3-methyl-cholanthrene of functional and structural changes in rat liver due to carbon tetrachloride inhalation. J Pharmacol Exp Ther. 1975 Apr;193(1):281–292. [PubMed] [Google Scholar]
- Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
- St John T. P., Davis R. W. The organization and transcription of the galactose gene cluster of Saccharomyces. J Mol Biol. 1981 Oct 25;152(2):285–315. doi: 10.1016/0022-2836(81)90244-8. [DOI] [PubMed] [Google Scholar]
- Temin H. M., Kassner V. K. Replication of reticuloendotheliosis viruses in cell culture: acute infection. J Virol. 1974 Feb;13(2):291–297. doi: 10.1128/jvi.13.2.291-297.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Torchia T. E., Hopper J. E. Genetic and molecular analysis of the GAL3 gene in the expression of the galactose/melibiose regulon of Saccharomyces cerevisiae. Genetics. 1986 Jun;113(2):229–246. doi: 10.1093/genetics/113.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Trowbridge R. S., Lehmann J., Torchio C., Brophy P. Visna virus synthesized in absence of host-cell division and DNA synthesis. Microbios. 1980;29(116):71–80. [PubMed] [Google Scholar]
- Van Arsdell S. W., Stetler G. L., Thorner J. The yeast repeated element sigma contains a hormone-inducible promoter. Mol Cell Biol. 1987 Feb;7(2):749–759. doi: 10.1128/mcb.7.2.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Varmus H. E., Padgett T., Heasley S., Simon G., Bishop J. M. Cellular functions are required for the synthesis and integration of avian sarcoma virus-specific DNA. Cell. 1977 Jun;11(2):307–319. doi: 10.1016/0092-8674(77)90047-2. [DOI] [PubMed] [Google Scholar]
- Xu H., Boeke J. D. Inhibition of Ty1 transposition by mating pheromones in Saccharomyces cerevisiae. Mol Cell Biol. 1991 May;11(5):2736–2743. doi: 10.1128/mcb.11.5.2736. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yeh C. T., Wong S. W., Fung Y. K., Ou J. H. Cell cycle regulation of nuclear localization of hepatitis B virus core protein. Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6459–6463. doi: 10.1073/pnas.90.14.6459. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zack J. A., Arrigo S. J., Weitsman S. R., Go A. S., Haislip A., Chen I. S. HIV-1 entry into quiescent primary lymphocytes: molecular analysis reveals a labile, latent viral structure. Cell. 1990 Apr 20;61(2):213–222. doi: 10.1016/0092-8674(90)90802-l. [DOI] [PubMed] [Google Scholar]