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Microbiology 147 (2001), 1805-1813
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Microbiology (2001), 147, 1805-1813.
© 2001 Society for General Microbiology

Genetics and Molecular Biology

Identification of genes involved in the activation of the Bacillus thuringiensis inhA metalloprotease gene at the onset of sporulation

Cosette Grandvaleta,1, Myriam Gominet1 and Didier Lereclus1,2

Unité de Biochimie Microbienne, Centre National de la Recherche Scientifique URA 2172, Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris Cedex 15, France1
Station de Recherche de Lutte Biologique, Institut National de la Recherche Agronomique, La Minière, 78285 Guyancourt, France2

Author for correspondence: Didier Lereclus. Tel: +33 1 45 68 88 13. Fax: +33 1 45 68 89 38. e-mail: lereclus{at}pasteur.fr

The immune inhibitor A (InhA) metalloprotease from Bacillus thuringiensis specifically cleaves antibacterial proteins produced by the insect host, suggesting that it may contribute to the overall virulence of B. thuringiensis. The transcriptional regulation of the inhA gene in both B. thuringiensis and Bacillus subtilis was investigated. Using a transcriptional inhA'–lacZ fusion, it was shown that inhA expression is activated at the onset of sporulation. However, the transcriptional start site of inhA is similar to {sigma}A-dependent promoters, and deletion of the sporulation-specific sigma factors {sigma}F or {sigma}E had no effect on inhA expression in B. subtilis. The DNA region upstream from inhA contains two genes encoding polypeptides similar to the SinI and SinR regulators of B. subtilis. SinR is a DNA-binding protein regulating gene expression and SinI inhibits SinR activity. Overexpression of the sin genes affects the expression of the inhA'–lacZ transcriptional fusion in B. thuringiensis: early induction of inhA expression was observed when sinI was overexpressed, whereas inhA expression was reduced in a strain overexpressing sinR, suggesting that inhA transcription is repressed, directly or indirectly, by SinR. inhA transcription was greatly reduced in B. thuringiensis and B. subtilis spo0A mutants. Analysis of the inhA'–lacZ expression in abrB and abrB–spo0A mutants of B. subtilis indicates that the Spo0A-dependent regulation of inhA expression depends on AbrB, which is known to regulate expression of transition state and sporulation genes in B. subtilis.

Keywords: transcriptional regulation, protease, AbrB, Spo0A, Sin

The GenBank/EMBL/DDBJ accession number for the sequence reported in this paper is AF287346.

a Present address: ENSBANA, Département de Microbiologie, 1 Esplanade Erasme, 21000 Dijon, France.




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