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J Bacteriol, July 1998, p. 3548-3555, Vol. 180, No. 14
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Dual Promoters Are Responsible for Transcription Initiation of the fla/che Operon in Bacillus subtilis

W. Estacio, S. Santa Anna-Arriola, M. Adedipe, and L. M. Márquez-Magaña*

Department of Biology, San Francisco State University, San Francisco, California 94132

Received 30 January 1998/Accepted 11 May 1998

The fla/che region contains more than 30 genes required for flagellar synthesis and chemotaxis in Bacillus subtilis, including the gene for the flagellum-specific sigma D factor, sigD. Sequence and primer extension data demonstrate that a PA promoter immediately upstream of flgB, henceforth referred to as the fla/che PA, and the PD-3 promoter are active in vivo. Transcription from the PD-3 element is dependent on sigma D activity and is regulated by the flagellum-specific negative regulator, FlgM. In a strain containing a deletion of fla/che PA (PADelta ), sigma D protein was not detected, demonstrating that the fla/che PA is necessary for wild-type expression of the sigD gene. Thus, sigD is part of the >26-kb fla/che operon. Consistent with a lack of detectable sigma D protein, the PADelta strain grows as long filaments and does not express a sigma D-dependent hag::lacZ reporter construct. These phenotypes are indicative of a lack of sigD expression or complete inhibition of sigma D activity by FlgM. However, sigma D activity is found in a double mutant containing the PADelta and a null mutation in flgM. The double mutant no longer grows as long filaments, and expression of hag::lacZ is partially restored. These data demonstrate that a low level of sigma D activity does exist in the PADelta mutant but can be detected only in the presence of a null mutation in flgM. Therefore, normal expression of sigD may also involve another promoter(s) within the fla/che operon.

* Corresponding author. Mailing address: Department of Biology, San Francisco State University, 1600 Holloway Ave., San Francisco, CA 94132. Phone: (415) 338-3289. Fax: (415) 338-0927. E-mail: marquez{at}sfsu.edu.

J Bacteriol, July 1998, p. 3548-3555, Vol. 180, No. 14
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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