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Transcriptional activation of theBacillus subtilis ackA gene, encoding acetate kinase, was previously shown to require catabolite control protein A (CcpA) and sequences upstream of theackA promoter. CcpA, which is responsible for catabolite repression of a number of secondary carbon source utilization genes inB. subtilis and other gram-positive bacteria, recognizes acis -acting consensus sequence, designatedcre (catabolite response element), generally located within or downstream of the promoter of the repressed gene. Two sites resembling this sequence are centered at positions −116.5 and −56.5 of theackA promoter and have been termedcre1 andcre2 , respectively. Synthesis of acetate kinase, which is involved in the conversion of acetyl coenzyme A to acetate, is induced when cells are grown in the presence of an easily metabolized carbon source such as glucose. In this study,cre2 , the site closer to the promoter, and the region upstream ofcre2 were shown to be indispensable for CcpA-dependent transcriptional activation ofackA , whereascre1 was not required. In addition, insertion of 5 bp betweencre2 and the promoter disrupted activation, while 10 bp was tolerated, suggesting face-of-the-helix dependence of the position ofcre2 and/or upstream sequences. DNase footprinting experiments demonstrated binding of CcpA in vitro tocre2 but notcre1 , consistent with the genetic data. Activation ofackA transcription was blocked in aptsH1/crh double mutant, suggesting involvement of this pathway in CcpA-mediated transcriptional activation.

Transcriptional Activation of the Bacillus subtilis ackA Gene Requires Sequences Upstream of the Promoter