Abstract
A protein factor with an estimated molecular mass of 50 kDa has been purified to homogeneity from the silk gland of Bombyx mori. In the presence of a molar excess of this factor and eukaryotic DNA topoisomerase II, relaxed circular DNA is converted to the negatively supercoiled form. Eukaryotic DNA topoisomerase I cannot substitute for eukaryotic DNA topoisomerase II in the supercoiling reaction. The reaction is dependent on ATP and is inhibited by VP-16, a specific inhibitor of eukaryotic DNA topoisomerase II. When DNA topoisomerase I is subsequently added to the supercoiling reaction mixture, the supercoiled DNA becomes relaxed. These results suggest that when both the 50-kDa protein and eukaryotic DNA topoisomerase II are present in excess, unconstrained negative supercoils are introduced into DNA.
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.
- Baldi M. I., Benedetti P., Mattoccia E., Tocchini-Valentini G. P. In vitro catenation and decatenation of DNA and a novel eucaryotic ATP-dependent topoisomerase. Cell. 1980 Jun;20(2):461–467. doi: 10.1016/0092-8674(80)90632-7. [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]
- Briggs M. R., Kadonaga J. T., Bell S. P., Tjian R. Purification and biochemical characterization of the promoter-specific transcription factor, Sp1. Science. 1986 Oct 3;234(4772):47–52. doi: 10.1126/science.3529394. [DOI] [PubMed] [Google Scholar]
- Brill S. J., Sternglanz R. Transcription-dependent DNA supercoiling in yeast DNA topoisomerase mutants. Cell. 1988 Jul 29;54(3):403–411. doi: 10.1016/0092-8674(88)90203-6. [DOI] [PubMed] [Google Scholar]
- Gellert M. DNA topoisomerases. Annu Rev Biochem. 1981;50:879–910. doi: 10.1146/annurev.bi.50.070181.004311. [DOI] [PubMed] [Google Scholar]
- Giaever G. N., Wang J. C. Supercoiling of intracellular DNA can occur in eukaryotic cells. Cell. 1988 Dec 2;55(5):849–856. doi: 10.1016/0092-8674(88)90140-7. [DOI] [PubMed] [Google Scholar]
- Hirose S., Suzuki Y. In vitro transcription of eukaryotic genes is affected differently by the degree of DNA supercoiling. Proc Natl Acad Sci U S A. 1988 Feb;85(3):718–722. doi: 10.1073/pnas.85.3.718. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirose S., Tabuchi H., Yoshinaga K. GTP induces knotting, catenation, and relaxation of DNA by stoichiometric amounts of DNA topoisomerase II from Bombyx mori and HeLa cells. J Biol Chem. 1988 Mar 15;263(8):3805–3810. [PubMed] [Google Scholar]
- Hirose S., Tsuda M., Suzuki Y. Enhanced transcription of fibroin gene in vitro on covalently closed circular templates. J Biol Chem. 1985 Sep 5;260(19):10557–10562. [PubMed] [Google Scholar]
- Hsieh T. Knotting of the circular duplex DNA by type II DNA topoisomerase from Drosophila melanogaster. J Biol Chem. 1983 Jul 10;258(13):8413–8420. [PubMed] [Google Scholar]
- Hsieh T. Purification and properties of type II DNA topoisomerase from embryos of Drosophila melanogaster. Methods Enzymol. 1983;100:161–170. doi: 10.1016/0076-6879(83)00052-x. [DOI] [PubMed] [Google Scholar]
- Hübscher U., Stalder H. P. Mammalian DNA helicase. Nucleic Acids Res. 1985 Aug 12;13(15):5471–5483. doi: 10.1093/nar/13.15.5471. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Laskey R. A., Honda B. M., Mills A. D., Finch J. T. Nucleosomes are assembled by an acidic protein which binds histones and transfers them to DNA. Nature. 1978 Oct 5;275(5679):416–420. doi: 10.1038/275416a0. [DOI] [PubMed] [Google Scholar]
- Lee M. P., Sander M., Hsieh T. Nuclease protection by Drosophila DNA topoisomerase II. Enzyme/DNA contacts at the strong topoisomerase II cleavage sites. J Biol Chem. 1989 Dec 25;264(36):21779–21787. [PubMed] [Google Scholar]
- Liu L. F., Wang J. C. DNA-DNA gyrase complex: the wrapping of the DNA duplex outside the enzyme. Cell. 1978 Nov;15(3):979–984. doi: 10.1016/0092-8674(78)90281-7. [DOI] [PubMed] [Google Scholar]
- Liu L. F., Wang J. C. Micrococcus luteus DNA gyrase: active components and a model for its supercoiling of DNA. Proc Natl Acad Sci U S A. 1978 May;75(5):2098–2102. doi: 10.1073/pnas.75.5.2098. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liu L. F., Wang J. C. Supercoiling of the DNA template during transcription. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7024–7027. doi: 10.1073/pnas.84.20.7024. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sinden R. R., Carlson J. O., Pettijohn D. E. Torsional tension in the DNA double helix measured with trimethylpsoralen in living E. coli cells: analogous measurements in insect and human cells. Cell. 1980 Oct;21(3):773–783. doi: 10.1016/0092-8674(80)90440-7. [DOI] [PubMed] [Google Scholar]
- Stahl H., Dröge P., Knippers R. DNA helicase activity of SV40 large tumor antigen. EMBO J. 1986 Aug;5(8):1939–1944. doi: 10.1002/j.1460-2075.1986.tb04447.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tabuchi H., Hirose S. DNA supercoiling facilitates formation of the transcription initiation complex on the fibroin gene promoter. J Biol Chem. 1988 Oct 25;263(30):15282–15287. [PubMed] [Google Scholar]
- Tsuda M., Suzuki Y. Faithful transcription initiation of fibroin gene in a homologous cell-free system reveals an enhancing effect of 5' flanking sequence far upstream. Cell. 1981 Nov;27(1 Pt 2):175–182. doi: 10.1016/0092-8674(81)90371-8. [DOI] [PubMed] [Google Scholar]
- Wang J. C. DNA topoisomerases. Annu Rev Biochem. 1985;54:665–697. doi: 10.1146/annurev.bi.54.070185.003313. [DOI] [PubMed] [Google Scholar]
- Weintraub H. Assembly and propagation of repressed and depressed chromosomal states. Cell. 1985 Oct;42(3):705–711. doi: 10.1016/0092-8674(85)90267-3. [DOI] [PubMed] [Google Scholar]