We have studied the formation of open complexes between purified RNA polymerase from Escherichia coli and DNA fragments carrying the galP1 promoter, a promoter with an extended -10 region. Unusually, these complexes are formed readily at low temperatures. This low-temperature opening is unaffected by deletions of either upstream or downstream promoter sequences. We conclude that low-temperature open-complex formation is due to specific base sequences in and just upstream of the extended -10 region. In contrast, open complexes are not formed at low temperatures with DNA fragments carrying the E. coli cysG promoter, which also has an extended -10 region. This demonstrates that an extended -10 sequence alone is not sufficient for low-temperature opening. Additionally, we report the temperature dependence of a hybrid galP1–cysG promoter, the related galP2 and galP3 promoters and a derivative of galP1 with an improved -10 hexamer sequence.
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July 1996
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Research Article|
July 01 1996
Temperature-dependence of open-complex formation at two Escherichia coli promoters with extended −10 sequences
Helen D. BURNS;
Helen D. BURNS
1School of Biochemistry, University of Birmingham, P.O. Box 363, Birmingham B15 2TT, U.K.
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Tamara A. BELYAEVA;
Tamara A. BELYAEVA
1School of Biochemistry, University of Birmingham, P.O. Box 363, Birmingham B15 2TT, U.K.
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Stephen J. W. BUSBY;
Stephen J. W. BUSBY
*
*To whom correspondence should be addressed.
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Stephen D. MINCHIN
Stephen D. MINCHIN
1School of Biochemistry, University of Birmingham, P.O. Box 363, Birmingham B15 2TT, U.K.
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Publisher: Portland Press Ltd
Received:
January 18 1996
Revision Received:
March 13 1996
Accepted:
March 21 1996
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1996
1996
Biochem J (1996) 317 (1): 305–311.
Article history
Received:
January 18 1996
Revision Received:
March 13 1996
Accepted:
March 21 1996
Citation
Helen D. BURNS, Tamara A. BELYAEVA, Stephen J. W. BUSBY, Stephen D. MINCHIN; Temperature-dependence of open-complex formation at two Escherichia coli promoters with extended −10 sequences. Biochem J 1 July 1996; 317 (1): 305–311. doi: https://doi.org/10.1042/bj3170305
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