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Genes controlled by the essential YycG/YycF two‐component system of Bacillus subtilis revealed through a novel hybrid regulator approach
Author(s) -
Howell Alistair,
Dubrac Sarah,
Andersen Kasper Krogh,
Noone David,
Fert Juliette,
Msadek Tarek,
Devine Kevin
Publication year - 2003
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1046/j.1365-2958.2003.03661.x
Subject(s) - regulon , biology , bacillus subtilis , response regulator , operon , genetics , gene , consensus sequence , regulator , two component regulatory system , regulation of gene expression , peptide sequence , escherichia coli , bacteria , mutant
Summary The YycG/YycF two‐component system, originally identified in Bacillus subtilis , is very highly conserved and appears to be specific to low G + C Gram‐positive bacteria. This system is required for cell viability, although the basis for this and the nature of the YycF regulon remained elusive. Using a combined hybrid regulator/transcriptome approach involving the inducible expression of a PhoP′‐′YycF chimerical protein in B . subtilis , we have shown that expression of yocH , which encodes a potential autolysin, is specifically activated by YycF. Gel mobility shift and DNase I footprinting assays were used to show direct binding in vitro of purified YycF to the regulatory regions of yocH as well as ftsAZ , previously reported to be controlled by YycF. Nucleotide sequence analysis and site‐directed mutagenesis allowed us to define a potential consensus recognition sequence for the YycF response regulator, composed of two direct repeats: 5′‐TGT A/T A A/T/C‐N 5 ‐TGT A/T A A/T/C‐3′. A DNA‐motif analysis indicates that there are potentially up to 10 genes within the B . subtilis YycG/YycF regulon, mainly involved in cell wall metabolism and membrane protein synthesis. Among these, YycF was shown to bind directly to the region upstream from the ykvT gene, encoding a potential cell wall hydrolase, and the intergenic region of the tagAB / tagDEF divergon, encoding essential components of teichoic acid biosynthesis. Definition of a potential YycF recognition sequence allowed us to identify likely members of the YycF regulon in other low G + C Gram‐positive bacteria, including several pathogens such as Listeria monocytogenes , Staphylococcus aureus and Streptococcus pneumoniae .