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Science 14 September 1990:
Vol. 249. no. 4974, pp. 1261 - 1265
DOI: 10.1126/science.2399463
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Articles

Science, Vol 249, Issue 4974, 1261-1265
Copyright © 1990 by American Association for the Advancement of Science

articles

Template supercoiling by a chimera of yeast GAL4 protein and phage T7 RNA polymerase

EA Ostrander, P Benedetti, and JC Wang

Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.

Fusion of the DNA-binding domain of yeast GAL4 protein to the amino terminus of bacteriophage T7 RNA polymerase yields a chimera that retains the characteristics of its components. The presence of the GAL4 peptide allows the chimeric enzyme to anchor itself on the DNA template, and this anchoring in turn drives the formation of a supercoiled DNA loop, in linear or circular templates, when RNA synthesis at the polymerase site forces a translocation of the DNA relative to the site. Nonspecific interaction between the chimeric enzyme and DNA appears to be sufficient to effect supercoiling during transcription. Transcription by the chimeric polymerase is strictly dependent on the presence of a T7 promoter; thus it provides a tool in vitro and in vivo for specifically supercoiling DNA segments containing T7 promoter sequences.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Multiple Roles for the T7 Promoter Nontemplate Strand during Transcription Initiation and Polymerase Release.
Q. Guo and R. Sousa (2005)
J. Biol. Chem. 280, 3474-3482
   Abstract »    Full Text »    PDF »
DNA communications by Type III restriction endonucleases--confirmation of 1D translocation over 3D looping.
L. J. Peakman and M. D. Szczelkun (2004)
Nucleic Acids Res. 32, 4166-4174
   Abstract »    Full Text »    PDF »
How do site-specific DNA-binding proteins find their targets?.
S. E. Halford and J. F. Marko (2004)
Nucleic Acids Res. 32, 3040-3052
   Abstract »    Full Text »    PDF »
Exposure of T7 RNA Polymerase to the Isolated Binding Region of the Promoter Allows Transcription from a Single-stranded Template.
A. Kukarin, M. Rong, and W. T. McAllister (2003)
J. Biol. Chem. 278, 2419-2424
   Abstract »    Full Text »    PDF »
In vitro transcription of a torsionally constrained template.
T. Bentin and P. E. Nielsen (2002)
Nucleic Acids Res. 30, 803-809
   Abstract »    Full Text »    PDF »
Role of Bacillus subtilis SpoIIIE in DNA Transport Across the Mother Cell-Prespore Division Septum.
J. Bath, L. J. Wu, J. Errington, and J. C. Wang (2000)
Science 290, 995-997
   Abstract »    Full Text »
In Vitro Studies on the Maintenance of Transcription-induced Stress by Histones and Polyamines.
H. F. Peng and V. Jackson (2000)
J. Biol. Chem. 275, 657-668
   Abstract »    Full Text »    PDF »
Tethering a Type IB Topoisomerase to a DNA Site by Enzyme Fusion to a Heterologous Site-selective DNA-binding Protein Domain.
G. L. Beretta, M. Binaschi, E. Zagni, L. Capuani, and G. Capranico (1999)
Cancer Res. 59, 3689-3697
   Abstract »    Full Text »    PDF »
Chirality of DNA trefoils: Implications in intramolecular synapsis of distant DNA segments.
S. Y. Shaw and J. C. Wang (1997)
PNAS 94, 1692-1697
   Abstract »    Full Text »    PDF »


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Science. ISSN 0036-8075 (print), 1095-9203 (online)