title: The dksA promoter is negatively feedback regulated by DksA and ppGpp.
abstract: Escherichia coli DksA is an RNA polymerase (RNAP)-binding protein required for the regulation of a number of promoters, including those for the biosynthesis of rRNA, many ribosomal proteins, components of the flagellum, and several amino acids. Previous work demonstrated that DksA protein levels do not vary greatly in different growth conditions. We show here that DksA is a stable protein whose levels are kept constant by a negative feedback loop in which transcription from the dksA promoter is inhibited by DksA protein in conjunction with its cofactor ppGpp. We map the primary dksA promoter by primer extension, show that its activity increases in a strain lacking DksA, that the DksA protein accumulates when expressed from an exogenous promoter, that inhibition of transcription by DksA is direct since it occurs with purified components in vitro, and that inhibition correlates with effects of DksA on the lifetime of the dksA promoter complex. This work provides a mechanistic basis for the maintenance of constant cellular levels of DksA in E. coli and supports the model that transcription regulation by ppGpp/DksA derives from fluctuations in the concentrations of the small molecule cofactor rather than of DksA itself.
authors.affiliation: Department of Bacteriology, University of Wisconsin-Madison, 1550 Linden Drive, Madison, WI 53706, USA.
authors.forename: Pete
authors.initials: P
authors.lastname: Chandrangsu
authors.forename: Justin J
authors.initials: JJ
authors.lastname: Lemke
authors.forename: Richard L
authors.initials: RL
authors.lastname: Gourse
mesh_terms: D015536:Down-Regulation; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D006150:Guanine Nucleotides / Q000378:metabolism*; D011401:Promoter Regions, Genetic
chemical_list: D029968:Escherichia coli Proteins; D006150:Guanine Nucleotides; C090751:dksA protein, E coli; C018957:guanosine 5'-diphosphate-3'-monophosphate