Abstract
We have studied the binding of the tumor antigen (T-antigen) of simian virus 40 to simian virus 40 chromatin (minichromosomes). The minichromosomes isolated from infected cells by a modification of standard techniques were relatively free of contaminating RNA and cellular DNA and had a ratio (by weight) of protein to DNA of approximately 1; their DNA was 50 to 60% digestible to an acid-soluble form by staphylococcal nuclease. Cleavage of this chromatin with restriction endonucleases indicated that the nuclease-resistant regions were randomly distributed in the population of minichromosomes, but were not randomly distributed within minichromosomes. Only 20 to 35% of these minichromosomes adsorbed nonspecifically to nitrocellulose filters, permitting binding studies between simian virus 40 T-antigen and chromatin to be performed. Approximately two to three times as much T-antigen was required to bind chromatin as to bind an equivalent amount of free DNA. When T-antigen was present in excess, both chromatin and free DNA were quantitatively retained on the filters. On the other hand, when DNA or chromatin was present in excess, only one-third as much chromatin as DNA was retained. We suggest that T-antigen-chromatin complexes may be formed by the cooperative binding of T-antigen to chromatin, whereas T-antigen-DNA complexes may be formed by simple bimolecular interactions.
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Selected References
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- Axel R., Melchior W., Jr, Sollner-Webb B., Felsenfeld G. Specific sites of interaction between histones and DNA in chromatin. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4101–4105. doi: 10.1073/pnas.71.10.4101. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carroll R. B., Hager L., Dulbecco R. Simian virus 40 T antigen binds to DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3754–3757. doi: 10.1073/pnas.71.9.3754. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chou J. Y., Avila J., Martin R. G. Viral DNA synthesis in cells infected by temperature-sensitive mutants of simian virus 40. J Virol. 1974 Jul;14(1):116–124. doi: 10.1128/jvi.14.1.116-124.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Christiansen G., Landers T., Griffith J., Berg P. Characterization of components released by alkali disruption of simian virus 40. J Virol. 1977 Mar;21(3):1079–1084. doi: 10.1128/jvi.21.3.1079-1084.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clark R. J., Felsenfeld G. Association of arginine-rich histones with G-C-rich regions of DNA in chromatin. Nat New Biol. 1972 Dec 20;240(103):226–229. doi: 10.1038/newbio240226a0. [DOI] [PubMed] [Google Scholar]
- Clark R. J., Felsenfeld G. Structure of chromatin. Nat New Biol. 1971 Jan 27;229(4):101–106. doi: 10.1038/newbio229101a0. [DOI] [PubMed] [Google Scholar]
- Compton J. L., Bellard M., Chambon P. Biochemical evidence of variability in the DNA repeat length in the chromatin of higher eukaryotes. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4382–4386. doi: 10.1073/pnas.73.12.4382. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cremisi C., Pignatti P. F., Croissant O., Yaniv M. Chromatin-like structures in polyoma virus and simian virus 10 lytic cycle. J Virol. 1975 Jan;17(1):204–211. doi: 10.1128/jvi.17.1.204-211.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Danna K. J., Sack G. H., Jr, Nathans D. Studies of simian virus 40 DNA. VII. A cleavage map of the SV40 genome. J Mol Biol. 1973 Aug 5;78(2):363–376. doi: 10.1016/0022-2836(73)90122-8. [DOI] [PubMed] [Google Scholar]
- Fareed G. C., Garon G. F., Salzman N. P. Origin and direction of simian virus 40 deoxyribonucleic acid replication. J Virol. 1972 Sep;10(3):484–491. doi: 10.1128/jvi.10.3.484-491.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Griffith J. D. Chromatin structure: deduced from a minichromosome. Science. 1975 Mar 28;187(4182):1202–1203. doi: 10.1126/science.187.4182.1202. [DOI] [PubMed] [Google Scholar]
- Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
- Huang E. S., Estes M. K., Pagano J. S. Structure and function of the polypeptides in simian virus 40. I. Existence of subviral deoxynucleoprotein complexes. J Virol. 1972 Jun;9(6):923–929. doi: 10.1128/jvi.9.6.923-929.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jessel D., Hudson J., Landau T., Tenen D., Livingston D. M. Interaction of partially purified simian virus 40 T antigen with circular viral DNA molecules. Proc Natl Acad Sci U S A. 1975 May;72(5):1960–1964. doi: 10.1073/pnas.72.5.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jessel D., Landau T., Hudson J., Lalor T., Tenen D., Livingston D. M. Identification of regions of the SV40 genome which contain preferred SV40 T antigen-binding sites. Cell. 1976 Aug;8(4):535–545. doi: 10.1016/0092-8674(76)90222-1. [DOI] [PubMed] [Google Scholar]
- Jones O. W., Berg P. Studies on the binding of RNA polymerase to polynucleotides. J Mol Biol. 1966 Dec 28;22(2):199–209. doi: 10.1016/0022-2836(66)90126-4. [DOI] [PubMed] [Google Scholar]
- Kuchino T., Yamaguchi N. Characterization of T antigen in cells infected with a temperature-sensitive mutant of simian virus 40. J Virol. 1975 Jun;15(6):1302–1307. doi: 10.1128/jvi.15.6.1302-1307.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lee T. N., Brockman W. W., Nathans D. Evolutionary variants of simian virus 40: cloned substituted variants containing multiple initiation sites for DNA replication. Virology. 1975 Jul;66(1):53–69. doi: 10.1016/0042-6822(75)90178-6. [DOI] [PubMed] [Google Scholar]
- Livingston D. M., Henderson I. C., Hudson J. SV40 T antigen: partial purification and properties. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):283–289. doi: 10.1101/sqb.1974.039.01.037. [DOI] [PubMed] [Google Scholar]
- McCann P. P., Martin R. G. Nuclease digestion of the DNA-protein complex from simian virus 40 virions. Biochim Biophys Acta. 1975 Jul 7;395(3):280–283. doi: 10.1016/0005-2787(75)90198-7. [DOI] [PubMed] [Google Scholar]
- Osborn M., Weber K. SV40: T antigen, the A function and transformation. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):267–276. doi: 10.1101/sqb.1974.039.01.035. [DOI] [PubMed] [Google Scholar]
- Oxman M. N., Takemoto K. K., Eckhart W. Polyoma T antigen synthesis by temperature-sensitive mutants of polyoma virus. Virology. 1972 Sep;49(3):675–682. doi: 10.1016/0042-6822(72)90524-7. [DOI] [PubMed] [Google Scholar]
- Pett D. M., Estes M. K., Pagano J. S. Structural proteins of simian virus 40. I. Histone characteristics of low-molecular-weight polypeptides. J Virol. 1975 Feb;15(2):379–385. doi: 10.1128/jvi.15.2.379-385.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Polisky B., McCarthy B. Location of histones on simian virus 40 DNA. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2895–2899. doi: 10.1073/pnas.72.8.2895. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Riggs A. D., Suzuki H., Bourgeois S. Lac repressor-operator interaction. I. Equilibrium studies. J Mol Biol. 1970 Feb 28;48(1):67–83. doi: 10.1016/0022-2836(70)90219-6. [DOI] [PubMed] [Google Scholar]
- Sen A., Hancock R., Levine A. J. The properties and origin of the proteins in the SV40 nucleoprotein complex. Virology. 1974 Sep;61(1):11–21. doi: 10.1016/0042-6822(74)90237-2. [DOI] [PubMed] [Google Scholar]
- Shaw B. R., Herman T. M., Kovacic R. T., Beaudreau G. S., Van Holde K. E. Analysis of subunit organization in chicken erythrocyte chromatin. Proc Natl Acad Sci U S A. 1976 Feb;73(2):505–509. doi: 10.1073/pnas.73.2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sollner-Webb B., Felsenfeld G. A comparison of the digestion of nuclei and chromatin by staphylococcal nuclease. Biochemistry. 1975 Jul;14(13):2915–2920. doi: 10.1021/bi00684a019. [DOI] [PubMed] [Google Scholar]
- Tegtmeyer P. Simian virus 40 deoxyribonucleic acid synthesis: the viral replicon. J Virol. 1972 Oct;10(4):591–598. doi: 10.1128/jvi.10.4.591-598.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tenen D. G., Baygell P., Livingston D. M. Thermolabile T (tumor) antigen from cells transformed by a temperature-sensitive mutant of simian virus 40. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4351–4355. doi: 10.1073/pnas.72.11.4351. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tenen D. G., Martin R. G., Anderson J., Livingston D. M. Biological and biochemical studies of cells transformed by simian virus 40 temperature-sensitive gene A mutants and A mutant revertants. J Virol. 1977 Apr;22(1):210–218. doi: 10.1128/jvi.22.1.210-218.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Varshavsky A. J., Bakayev V. V., Chumackov P. M., Georgiev G. P. Minichromosome of simian virus 40: presence of histone HI. Nucleic Acids Res. 1976 Aug;3(8):2101–2113. doi: 10.1093/nar/3.8.2101. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vogel T., Singer M. The interaction of histones with simian virus 40 supercoiled circular deoxyribonucleic acid in vitro. J Biol Chem. 1975 Jan 25;250(2):796–798. [PubMed] [Google Scholar]
- Whitlock J. P., Jr, Simpson R. T. Removal of histone H1 exposes a fifty base pair DNA segment between nucleosomes. Biochemistry. 1976 Jul 27;15(15):3307–3314. doi: 10.1021/bi00660a022. [DOI] [PubMed] [Google Scholar]