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Journal of Bacteriology, November 1998, p. 5619-5625, Vol. 180, No. 21
Department of Chemistry and Biochemistry and
Molecular Biology Institute, University of California, Los Angeles,
Los Angeles, California 90095-1569
Received 26 May 1998/Accepted 26 August 1998
Sigma 54 associates with bacterial core RNA polymerase and converts
it into an enhancer-responsive enzyme. Deletion of the N-terminal 40 amino acids is known to result in loss of the ability to respond to
enhancer binding proteins. In this work PCR mutagenesis and genetic
screens were used to identify a small patch, from amino acids 33 to 37, that is required for proper response to activator in vivo.
Site-directed single point mutants within this segment were constructed
and studied. Two of these were defective in responding to the enhancer
binding protein in vitro. The mutants could still direct the polymerase
to bind to DNA and initiate transient melting. However, they failed in
directing activator-dependent formation of a heparin-stable open
complex. Thus, amino acid region 33 to 37 includes critical activation
response determinants. This region overlaps the larger leucine patch
negative-control region, suggesting that anti-inhibition and positive
activation are closely coupled events.
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Identification of an N-Terminal Region of Sigma 54 Required for Enhancer Responsiveness
*
Corresponding author. Mailing address: Department of
Chemistry & Biochemistry and Molecular Biology Institute, University of
California, Los Angeles, P.O. Box 951569, Los Angeles, CA 90095-1569. Phone: (310) 825-1620. Fax: (310) 267-2302. E-mail:
gralla{at}ewald.mbi.ucla.edu.
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