Transcriptional regulation of a promoter in the men gene cluster of Bacillus subtilis

The control of men gene expression during growth and sporulation of Bacillus subtilis was examined at the transcriptional level. Two different approaches were used. (i) Steady-state levels of men-specific mRNA were measured directly. (ii) A men'-lacZ gene fusion was constructed. In both cases, it was observed that men promoter activity was maximal at the onset of sporulation and declined soon thereafter. These kinetics were similar to the pattern of menaquinone accumulation previously observed. Expression from the men promoter was independent of the presence of the products of the spo0A and spo0H genes and was enhanced by addition of glucose and glutamine to the culture medium. DNA sequence analysis of the promoter region revealed a potential recognition site for the principal vegetative form of RNA polymerase but not for any of the known minor polymerase forms. The functionality in vivo of the promoter sequence was confirmed by high-resolution S1 nuclease mapping of the transcript start site. An additional sequence element was identified that is shared by the sdhA, citG, and ctaA promoters and may indicate a common regulatory mechanism in the expression of these genes.

[1]  T. Kudo,et al.  Cloning of a developmentally regulated element from alkalophilic Bacillus subtilis DNA , 1985, Journal of bacteriology.

[2]  K. Struhl,et al.  Saturation mutagenesis of the yeast his3 regulatory site: requirements for transcriptional induction and for binding by GCN4 activator protein. , 1986, Science.

[3]  J. S. Miles,et al.  Complete nucleotide sequence of the fumarase gene (citG) of Bacillus subtilis 168 , 1985, Nucleic Acids Res..

[4]  J. Messing New M13 vectors for cloning. , 1983, Methods in enzymology.

[5]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[6]  E. Chen,et al.  Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. , 1985, DNA.

[7]  Jeffrey H. Miller Experiments in molecular genetics , 1972 .

[8]  C. Saunders,et al.  Use of chromosomal integration in the establishment and expression of blaZ, a Staphylococcus aureus beta-lactamase gene, in Bacillus subtilis , 1984, Journal of bacteriology.

[9]  H. Birnboim,et al.  A rapid alkaline extraction procedure for screening recombinant plasmid DNA. , 1979, Nucleic acids research.

[10]  P. Schaeffer,et al.  Catabolic repression of bacterial sporulation. , 1965, Proceedings of the National Academy of Sciences of the United States of America.

[11]  A. Flavell,et al.  Nucleic acid hybridization: A practical approach: B.D. Hames and S.J. Higgins (editors) IRL Press Ltd., Oxford, UK, 256 pp., £14.00/US$25.00 (softbound), £22.00/US$40.00 (hardbound), ISBN 0-947946-23-3 (softbound), ISBN 0-947946-61-6 (hardbound) , 1987 .

[12]  R. Treisman,et al.  Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. , 1980, Methods in enzymology.

[13]  A. Sonenshein,et al.  Bacillus subtilis citB gene is regulated synergistically by glucose and glutamine , 1985, Journal of bacteriology.

[14]  J. Petit,et al.  Influence du taux de croissance sur la constitution du spectre hématinique de B. Subtilis , 1957 .

[15]  P. Fortnagel The regulation of aconitase and isocitrate dehydrogenase in sporulation mutants of Bacillus subtilis. , 1970, Biochimica et biophysica acta.

[16]  M. Arnaud,et al.  Observations on the regulation of the synthesis of the tricarboxylic acid cycle enzymes in Bacillus subtilis, Marburg , 1964 .

[17]  S. Farrand,et al.  Physiological Effects of Menaquinone Deficiency in Bacillus subtilis , 1973, Journal of bacteriology.

[18]  A. Sonenshein,et al.  Relationship between aconitase gene expression and sporulation in Bacillus subtilis , 1987, Journal of bacteriology.

[19]  P. Miller,et al.  Molecular cloning and preliminary genetic analysis of the men gene cluster of Bacillus subtilis , 1988, Journal of bacteriology.

[20]  J. Guest,et al.  Nucleotide sequence of the gene for cytochrome b558 of the Bacillus subtilis succinate dehydrogenase complex , 1986, Journal of bacteriology.

[21]  R. S. Hanson,et al.  Effect of Different Nutritional Conditions on the Synthesis of Tricarboxylic Acid Cycle Enzymes , 1967, Journal of bacteriology.

[22]  A. Sonenshein,et al.  Altered regulation of the glnA gene in glutamine synthetase mutants of Bacillus subtilis , 1986, Journal of bacteriology.

[23]  R. Doi,et al.  Multiple procaryotic ribonucleic acid polymerase sigma factors. , 1986, Microbiological reviews.

[24]  D. Ebbole,et al.  Organization and regulation of genes encoding biosynthetic enzymes in Bacillus subtilis. , 1988, The Journal of biological chemistry.

[25]  S. Farrand,et al.  Changes in Menaquinone Concentration During Growth and Early Sporulation in Bacillus subtilis , 1974, Journal of bacteriology.

[26]  R. Losick,et al.  Regulation of a promoter that is utilized by minor forms of RNA polymerase holoenzyme in Bacillus subtilis. , 1986, Journal of molecular biology.

[27]  R. Losick,et al.  Genetics of endospore formation in Bacillus subtilis. , 1986, Annual review of genetics.

[28]  S. Baumberg,et al.  Sequence analysis of the Bacillus subtilis argC promoter region. , 1986, Gene.