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Journal of Bacteriology, June 1999, p. 3351-3357, Vol. 181, No. 11
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Mutant Forms of Salmonella typhimurium sigma 54 Defective in Transcription Initiation but Not Promoter Binding Activity

Mary T. Kelly and Timothy R. Hoover*

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

Received 17 February 1999/Returned for modification 15 March 1999/Accepted 6 April 1999

Transcription initiation with sigma 54-RNA polymerase holoenzyme (sigma 54-holoenzyme) has absolute requirements for an activator protein and ATP hydrolysis. sigma 54's binding to core RNA polymerase and promoter DNA has been well studied, but little is known about its role in the subsequent steps of transcription initiation. Following random mutagenesis, we isolated eight mutant forms of Salmonella typhimurium sigma 54 that were deficient in transcription initiation but still directed sigma 54-holoenzyme to the promoter to form a closed complex. Four of these mutant proteins had amino acid substitutions in region I, which had been shown previously to be required for sigma 54-holoenzyme to respond to the activator. From the remaining mutants, we identified four residues in region III which when altered affect the function of sigma 54 at some point after closed-complex formation. These results suggest that in addition to its role in core and DNA binding, region III participates in one or more steps of transcription initiation that follow closed-complex formation.

* 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.

Journal of Bacteriology, June 1999, p. 3351-3357, Vol. 181, No. 11
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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