The spa-box for transcriptional activation of subtilin biosynthesis and immunity in Bacillus subtilis

Autor:	Karl-Dieter Entian, Bettina Himmel, Torsten Stein, Stefan Heinzmann, Peter Kiesau
Rok vydání:	2003
Předmět:	chemistry.chemical_classification biology Bacillus subtilis biology.organism_classification Microbiology Response regulator chemistry.chemical_compound chemistry Biosynthesis Biochemistry Immunity Gene cluster Nucleotide Spar Molecular Biology DNA
Zdroj:	Molecular Microbiology. 47:1627-1636
ISSN:	1365-2958 0950-382X
DOI:	10.1046/j.1365-2958.2003.03374.x
Popis:	Summary The subtilin gene cluster (spa) of Bacillus subtilis ATCC 6633 is organized in transcriptional units spaBTC, spaS, spaIFEG and spaRK. Specific binding of the response regulator protein SpaR to spaB, spaS and spaI DNA promoter fragments was shown by means of electromobility shift assays. A repeated pentanucleotide sequence spaced by six nucleotides was identified as SpaR binding motif (spa-box). Saturating mutational analysis of the spa-box by single- and multiple-base-pair substitutions revealed the consensus motif (A/T)TGAT for optimal SpaR binding with the second, third and fifth position being absolutely conservative. Variations in the spacer size between the two pentanucleotide repeats revealed a strong conservation of their relative location. Only DNA with a proximal arrangement of two pentanucleotide repeats showed affinity to SpaR. A 2:1 stoichiometry between SpaR and DNA was obtained by optical biosensor analyses, which corresponds to the binding of two SpaR proteins per spa-box.
Databáze:	OpenAIRE
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