J Bacteriol 1998 Jul;180(14):3548-55

Dual promoters are responsible for transcription initiation of the fla/che operon in Bacillus subtilis.

Estacio W, Anna-Arriola SS, Adedipe M, Marquez-Magana LM.

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

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 sigmaD 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 sigmaD activity and is regulated by the flagellum-specific negative regulator, FlgM. In a strain containing a deletion of fla/che PA (PADelta), sigmaD 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 sigmaD protein, the PADelta strain grows as long filaments and does not express a sigmaD-dependent hag::lacZ reporter construct. These phenotypes are indicative of a lack of sigD expression or complete inhibition of sigmaD activity by FlgM. However, sigmaD 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 sigmaD 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.

PMID: 9657996 [PubMed - indexed for MEDLINE]

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Márquez-Magaña Lab.
Department of Biology
SFSU, 1600 Holloway Avenue, San Francisco, CA 94132