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Journal of Bacteriology, May 2001, p. 2995-3003, Vol. 183, No. 10
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.10.2995-3003.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Forespore-Specific Transcription of the lonB Gene during Sporulation in Bacillus subtilis

Monica Serrano,1 Sven Hövel,2 Charles P. Moran Jr.,3 Adriano O. Henriques,1 and Uwe Völker2,*

Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, 2781-901 Oeiras Codex, Portugal1; Laboratorium für Mikrobiologie, Philipps-Universität and Max-Planck-Institut für Terrestrische Mikrobiologie, 35043 Marburg, Germany2; and Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia 303223

Received 27 November 2000/Accepted 8 February 2001

The Bacillus subtilis genome encodes two members of the Lon family of prokaryotic ATP-dependent proteases. One, LonA, is produced in response to temperature, osmotic, and oxidative stress and has also been implicated in preventing sigma G activity under nonsporulation conditions. The second is encoded by the lonB gene, which resides immediately upstream from lonA. Here we report that transcription of lonB occurs during sporulation under sigma F control and thus is restricted to the prespore compartment of sporulating cells. First, expression of a lonB-lacZ transcriptional fusion was abolished in strains unable to produce sigma F but remained unaffected upon disruption of the genes encoding the early and late mother cell regulators sigma E and sigma K or the late forespore regulator sigma G. Second, the fluorescence of strains harboring a lonB-gfp fusion was confined to the prespore compartment and depended on sigma F production. Last, primer extension analysis of the lonB transcript revealed -10 and -35 sequences resembling the consensus sequence recognized by sigma F-containing RNA polymerase. We further show that the lonB message accumulated as a single monocistronic transcript during sporulation, synthesis of which required sigma F activity. Disruption of the lonB gene did not confer any discernible sporulation phenotype to otherwise wild-type cells, nor did expression of lonB from a multicopy plasmid. In contrast, expression of a fusion of the lonB promoter to the lonA gene severely reduced expression of the sigma G-dependent sspE gene and the frequency of sporulation. In confirmation of earlier observations, we found elevated levels of sigma F-dependent activity in a spoIIIE47 mutant, in which the lonB region of the chromosome is not translocated into the prespore. Expression of either lonB or the PlonB-lonA fusion from a plasmid in the spoIIIE47 mutant reduced sigma F -dependent activity to wild-type levels. The results suggest that both LonA and LonB can prevent abnormally high sigma F activity but that only LonA can negatively regulate sigma G.

* Corresponding author. Mailing address: Laboratorium für Mikrobiologie, Philipps-Universität Marburg, Karl-von-Frisch-Str., 35032 Marburg, Germany. Phone: 49-6421-282-3478. Fax: 49-6421-282-8979. E-mail: voelkeru{at}mailer.uni-marburg.de.

Journal of Bacteriology, May 2001, p. 2995-3003, Vol. 183, No. 10
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.10.2995-3003.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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