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Journal of Bacteriology, November 2000, p. 6503-6508, Vol. 182, No. 22
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Transcription Initiation-Defective Forms of sigma 54 That Differ in Ability To Function with a Heteroduplex DNA Template

Mary T. Kelly,dagger John A. Ferguson III, and Timothy R. Hoover*

Department of Microbiology, University of Georgia, Athens, Georgia 30602

Received 14 August 2000/Accepted 6 September 2000

Transcription by sigma 54-RNA polymerase holoenzyme requires an activator that catalyzes isomerization of the closed promoter complex to an open complex. We examined mutant forms of Salmonella enterica serovar Typhimurium sigma 54 that were defective in transcription initiation but retained core RNA polymerase- and promoter-binding activities. Four of the mutant proteins allowed activator-independent transcription from a heteroduplex DNA template. One of these mutant proteins, L124P V148A, had substitutions in a sequence that had not been shown previously to participate in the prevention of activator-independent transcription. The remaining mutants did not allow efficient activator-independent transcription from the heteroduplex DNA template and had substitutions within a conserved 20-amino-acid segment (Leu-179 to Leu-199), suggesting a role for this sequence in transcription initiation.

* Corresponding author. Mailing address: Department of Microbiology, 527 Biological Sciences Building, University of Georgia, Athens, GA 30602. Phone: (706) 542-2675. Fax: (706) 542-2674. E-mail: trhoover{at}arches.uga.edu.

dagger Present address: Department of Biochemistry, Conway Institute of Biomolecular and Biomedical Sciences, University College Dublin, Belfield, Dublin 4, Ireland.

Journal of Bacteriology, November 2000, p. 6503-6508, Vol. 182, No. 22
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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