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Nucleic Acids Research, 1989, Vol. 17, No. 3 999-1017
© 1989

ENZYMOLOGY

Sigma-G RNA polymerise controls forespore-specific expression of the glucose dehaydrogenase operon in Bacillus subtilis

Yoshihiro Nakatani*, Wayne L. Nicholson1, Klaus-Dieter Neitzke, Peter Setlow1 and Ernst Freese

Laboratory of Molecular Biology, National Institute of Neurological Disorders and Stroke, National Institutes of Health Bethesda, MD 20892 1Department of Biochemistry, University of Connecticut Health Center Farmington, CT 06032, USA

*To whom correspondence should be addressed

Received November 1, 1988. Revised January 10, 1989. Accepted January 10, 1989.

The gene encoding glucose dehydrogenase (gdh) is part of an operon whose expression is transcriptionally activated specifically in the developing forespore of Bacillus subtilis at stage lll of sporulation. The in vivo startpoint of gdh transcription was determined using primer extension analysis. Deletion mapping and site-specific mutagenesis experiments indicated that the region responsible for regulated expression of gdh in vivo was limited to the "-35" and "-10" regions preceding the transcriptional start site. RNA polymerase containing {sigma}G (E{sigma}G) transcribed gdh in vitro with a start site identical to that found in vivo, and transcription of gdh by E{sigma}G in vitro also did not require any specific sequences upstream from "-35" region. These results suggest that the appearance of E{sigma}G in the forespore at stage III of sporulation is sufficient to cause temporal and compartment-specific expression of the gdh operon.


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