The -16 region of Bacillus subtilis and other gram-positive bacterial promoters.
The Bacillus subtilis alpha-amylase promoter amy P 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 amy P by both B.subtilis and Escherichia coli RNA polymerase (RNAP). A second alpha-amylase promoter ( amy P2) 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 A n and T n 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 alpha-amylase promoters, necessary for transcription by the major form of B.subtilis and E.coli RNAP.
Department of Bacteriology, University of Wisconsin-Madison, E. B. Fred Hall, Madison, WI 53706, USA.
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Voskuil
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Chambliss
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence; D003352:Corynebacterium / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D006094:Gram-Positive Bacteria / Q000235:genetics; D007778:Lactobacillus / Q000235:genetics; D013294:Lactococcus lactis / Q000235:genetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011401:Promoter Regions, Genetic; D012808:Sigma Factor / Q000378:metabolism; D013296:Streptococcus pneumoniae / Q000235:genetics; D014158:Transcription, Genetic; D000516:alpha-Amylases / Q000235:genetics*
D001426:Bacterial Proteins; D012808:Sigma Factor; C081775:RNA polymerase sigma 70; D012321:DNA-Directed RNA Polymerases; D000516:alpha-Amylases