Identification of an N-terminal region of sigma 54 required for enhancer responsiveness.

Abstract:

: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.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Syed A,Gralla JD

doi

10.1128/JB.180.21.5619-5625.1998

subject

Has Abstract

pub_date

1998-11-01 00:00:00

pages

5619-25

issue

21

eissn

0021-9193

issn

1098-5530

journal_volume

180

pub_type

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