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Altering the level and regulation of the major sigma subunit of RNA polymerase affects gene expression and development in Bacillus subtilis

Authors

  • Karen A. Hicks,

    1. Department of Biology, Building 68–530, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
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    • Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403, USA.

  • Alan D. Grossman

    Corresponding author
    1. Department of Biology, Building 68–530, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
    • *For correspondence. E-mail adg@mit.edu; Tel. (617) 253 1515; Fax (617) 253 8699.

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Summary

In Bacillus subtilis, the major sigma factor, sigma-A (rpoD), and the minor sigma factor, sigma-H (spo0H), are present during growth and are required for the initiation of sporulation. Our experiments indicate that sigma-A and sigma-H compete for binding to core RNA polymerase. We used a fusion of rpoD to the Lacl-repressible IPTG-inducible promoter, Pspac, to vary the levels of sigma-A in the cell. Increasing the amount of sigma-A caused a decrease in expression of genes controlled by sigma-H, and a delay in the production of heat-resistant spores. Decreasing the amount of sigma-A, in a strain deleted for the chromosomal rpoD, caused an increase in expression of genes controlled by sigma-H. As rpoD itself is controlled by at least two promoters recognized by RNA polymerase that contains sigma-H, the effect of sigma-A levels on expression of sigma-H-controlled promoters represents a feedback mechanism that might contribute to maintaining appropriate levels of sigma-A. While the level of sigma-A was important for efficient sporulation, our results indicate that the normal transcriptional control of rpoD, in the context of the rpoD operon and the numerous promoters in that operon, is not required for efficient sporulation or germination, provided that the sigma-A level from a heterologous promoter is comparable to that in wild-type cells.

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