Abstract
The kinetics of open complex formation were measured by migration retardation assay and DNase I footprinting at the activator-dependent promoters ara P1, lac P1 and gal P1. In each case, the rate of open complex formation was significantly faster if the activator, AraC for ara and CAP for lac and gal, had been added before RNA polymerase. The results indicate that complexes of transcriptional activators, RNA polymerase and promoter can exist in two states, one which can form open complexes rapidly and one which cannot.
Full Text
The Full Text of this article is available as a PDF (65.9 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Donnelly C. E., Reznikoff W. S. Mutations in the lac P2 promoter. J Bacteriol. 1987 May;169(5):1812–1817. doi: 10.1128/jb.169.5.1812-1817.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Galas D. J., Schmitz A. DNAse footprinting: a simple method for the detection of protein-DNA binding specificity. Nucleic Acids Res. 1978 Sep;5(9):3157–3170. doi: 10.1093/nar/5.9.3157. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hahn S., Hendrickson W., Schleif R. Transcription of Escherichia coli ara in vitro. The cyclic AMP receptor protein requirement for PBAD induction that depends on the presence and orientation of the araO2 site. J Mol Biol. 1986 Apr 5;188(3):355–367. doi: 10.1016/0022-2836(86)90160-9. [DOI] [PubMed] [Google Scholar]
- Hendrickson W., Schleif R. F. Regulation of the Escherichia coli L-arabinose operon studied by gel electrophoresis DNA binding assay. J Mol Biol. 1984 Sep 25;178(3):611–628. doi: 10.1016/0022-2836(84)90241-9. [DOI] [PubMed] [Google Scholar]
- Herbert M., Kolb A., Buc H. Overlapping promoters and their control in Escherichia coli: the gal case. Proc Natl Acad Sci U S A. 1986 May;83(9):2807–2811. doi: 10.1073/pnas.83.9.2807. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lavigne M., Kolb A., Buc H. Transcription activation by cAMP receptor protein (CRP) at the Escherichia coli gal P1 promoter. Crucial role for the spacing between the CRP binding site and the -10 region. Biochemistry. 1992 Oct 13;31(40):9647–9656. doi: 10.1021/bi00155a018. [DOI] [PubMed] [Google Scholar]
- Meiklejohn A. L., Gralla J. D. Entry of RNA polymerase at the lac promoter. Cell. 1985 Dec;43(3 Pt 2):769–776. doi: 10.1016/0092-8674(85)90250-8. [DOI] [PubMed] [Google Scholar]
- Reeder T., Schleif R. AraC protein can activate transcription from only one position and when pointed in only one direction. J Mol Biol. 1993 May 20;231(2):205–218. doi: 10.1006/jmbi.1993.1276. [DOI] [PubMed] [Google Scholar]
- Schleif R. F., Favreau M. A. Hyperproduction of araC protein from Escherichia coli. Biochemistry. 1982 Feb 16;21(4):778–782. doi: 10.1021/bi00533a031. [DOI] [PubMed] [Google Scholar]
- Spassky A., Busby S., Buc H. On the action of the cyclic AMP-cyclic AMP receptor protein complex at the Escherichia coli lactose and galactose promoter regions. EMBO J. 1984 Jan;3(1):43–50. doi: 10.1002/j.1460-2075.1984.tb01759.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhang X., Reeder T., Schleif R. Transcription activation parameters at ara pBAD. J Mol Biol. 1996 Apr 26;258(1):14–24. doi: 10.1006/jmbi.1996.0230. [DOI] [PubMed] [Google Scholar]