The −16 region of Bacillus subtilis and other gram-positive bacterial promoters

• Published in: Nucleic acids research Vol. 26; no. 15; pp. 3584 - 3590
• Main Authors: Voskuil, Martin I., Chambliss, Glenn H.
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01-08-1998
• Subjects: alpha-Amylases - genetics; Bacillus subtilis; Bacillus subtilis - enzymology; Bacillus subtilis - genetics; Bacterial Proteins - metabolism; Base Sequence; Corynebacterium - genetics; DNA-Directed RNA Polymerases - metabolism; Escherichia coli; Gram-Positive Bacteria - genetics; Lactobacillus - genetics; Lactococcus lactis - genetics; Molecular Sequence Data; Mutagenesis; Promoter Regions, Genetic; Sigma Factor - metabolism; Streptococcus pneumoniae - genetics; Transcription, Genetic
• ISSN: 0305-1048; 1362-4962; 1362-4962
• DOI: 10.1093/nar/26.15.3584

The Bacillus subtilis α-amylase promoter amyP contains an essential TGTG motif (−16 region) upstream of the −10 region. Mutations of this region significantly reduced in vitro promoter strength. A −15 G→C transversion eliminated transcription from amyP by both B.subtilis and Escherichia coli RNA polymerase (RNAP). A second α-amylase promoter (amyP2) also required the −16 region for function. To determine conserved sequences in promoters containing −16 region elements, sequences of 64 B.subtilis promoters with the second TG motif of the −16 region were aligned and analyzed. Unlike the E.coli class of ‘extended −10 promoters’, with a similar TG motif but lacking a −35 region, the −16 region promoters contain highly conserved −35 regions. They also contain conserved An and Tn tracts upstream of the −35 region. In addition, we analyzed all available gram-positive bacterial promoter compilations to determine the generality of the −16 region. From this analysis, the −16 region TRTG motif (R = purine) appears to be a basic element found in a large portion of gram-positive bacterial promoters and is, in the case of at least the α-amylase promoters, necessary for transcription by the major form of B.subtilis and E.coli RNAP.