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Closed Access item Effect of mutations in the “extended −10” motif of three Bacillus subtilis σA-RNA polymerase-dependent promoters

Authors:Camacho, Ana
Salas, Margarita
Keywords:Bacillus subtilis, Bacteriophage ø29, Extended −10 motif, Promoters
Issue Date:26-Feb-1999
Citation:Journal of Molecular Biology 286(3): 683-693 (1999)
Abstract:The “extended −10“ motif described originally in Escherichia coli promoters occurs frequently in other bacterial promoters. Most Bacillus subtilis bacteriophage ø29 promoters contain this motif. To analyse the influence of the motif on σA-RNA polymerase transcription, the 5′-TG-3′ dinucleotide was changed to 5′-AC-3′ in three ø29 promoters. This change impaired RNA polymerase binding to the promoters; the yields of closed and open complexes were reduced independently of other differences inherent to each promoter. The mutation abolished transcription in vitro from a promoter lacking the consensus sequence at the −35 hexamer. In contrast, at other promoters with a −35 consensus sequence, the yield of run off transcription was not reduced by the mutation. Indeed an apparent interference phenomenon at high polymerase/DNA ratios was relieved. These results indicate that the extended −10 motif provides contact points for σA-RNA polymerase with a role restricted to the first steps of transcription.
Publisher version (URL):http://dx.doi.org/10.1006/jmbi.1998.2526
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