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Journal of Bacteriology, February 2004, p. 1078-1083, Vol. 186, No. 4
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.4.1078-1083.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

{alpha}-Helix E of Spo0A Is Required for {sigma}A- but Not for {sigma}H-Dependent Promoter Activation in Bacillus subtilis

Amrita Kumar,1 James A. Brannigan,2 and Charles P. Moran Jr.1*

Department of Microbiology & Immunology, Emory University School of Medicine, Atlanta, Georgia 30322,1 Department of Chemistry, Structural Biology Laboratory, University of York, Heslington, York, Y010 5YW, United Kingdom2

Received 2 July 2003/ Accepted 14 August 2003

At the onset of endospore formation in Bacillus subtilis, the DNA binding protein Spo0A activates transcription from two types of promoters. The first type includes the spoIIG and spoIIE promoters, which are used by {sigma}A-RNA polymerase, whereas the second type includes the spoIIA promoter, which is used by RNA polymerase containing the secondary sigma factor {sigma}H. Previous genetic analyses have identified specific amino acids in {alpha}-helix E of Spo0A that are important for activation of Spo0A-dependent, {sigma}A-dependent promoters. However, these amino acids are not required for activation of the {sigma}H-dependent spoIIA promoter. We now report the effects of additional single-amino-acid substitutions and the effects of deletions in {alpha}-helix E. The effects of alanine substitutions revealed one new position (239) in Spo0A that appears to be specifically required for activation of the {sigma}A-dependent promoters. Based on the effects of a deletion mutation, we suggest that {alpha}-helix E in Spo0A is not directly involved in interaction with {sigma}H-RNA polymerase.

* Corresponding author. Mailing address: Department of Microbiology & Immunology, Emory University School of Medicine, Atlanta, GA 30322. Phone: (404) 727-5969. Fax: (404) 727-3659. E-mail: moran{at}microbio.emory.edu.

Journal of Bacteriology, February 2004, p. 1078-1083, Vol. 186, No. 4
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.4.1078-1083.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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