title: pH-Dependent DNA Distortion and Repression of Gene Expression by Pectobacterium atrosepticum PecS.
abstract: Transcriptional activity is exquisitely sensitive to changes in promoter DNA topology. Transcription factors may therefore control gene activity by modulating the relative positioning of -10 and -35 promoter elements. The plant pathogen Pectobacterium atrosepticum, which causes soft rot in potatoes, must alter gene expression patterns to ensure growth in planta. In the related soft-rot enterobacterium Dickeya dadantii, PecS functions as a master regulator of virulence gene expression. Here, we report that P. atrosepticum PecS controls gene activity by altering promoter DNA topology in response to pH. While PecS binds the pecS promoter with high affinity regardless of pH, it induces significant DNA distortion only at neutral pH, the pH at which the pecS promoter is repressed in vivo. At pH ∼8, DNA distortions are attenuated, and PecS no longer represses the pecS promoter. A specific histidine (H142) located in a crevice between the dimerization- and DNA-binding regions is required for pH-dependent changes in DNA distortion and repression of gene activity, and mutation of this histidine renders the mutant protein incapable of repressing the pecS promoter. We propose that protonated PecS induces a DNA conformation at neutral pH in which -10 and -35 promoter elements are suboptimally positioned for RNA polymerase binding; on deprotonation of PecS, binding is no longer associated with significant changes in DNA conformation, allowing gene expression. We suggest that this mode of gene regulation leads to differential expression of the PecS regulon in response to alkalinization of the plant apoplast.
authors.affiliation: Department of Biological Sciences, Louisiana State University , Baton Rouge, Louisiana 70803, United States.
authors.forename: Dinesh K
authors.initials: DK
authors.lastname: Deochand
authors.affiliation: Department of Biological Sciences, Louisiana State University , Baton Rouge, Louisiana 70803, United States.
authors.forename: Jacob K
authors.initials: JK
authors.lastname: Meariman
authors.affiliation: Department of Biological Sciences, Louisiana State University , Baton Rouge, Louisiana 70803, United States.
authors.forename: Anne
authors.initials: A
authors.lastname: Grove
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D018744:DNA, Plant / Q000737:chemistry* / Q000378:metabolism; D018506:Gene Expression Regulation, Plant / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D009690:Nucleic Acid Conformation; D044043:Pectobacterium / Q000235:genetics / Q000502:physiology*; D011485:Protein Binding; D011198:Solanum tuberosum / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D018744:DNA, Plant