title: Genome-wide analysis of phosphorylated PhoP binding to chromosomal DNA reveals several novel features of the PhoPR-mediated phosphate limitation response in Bacillus subtilis.
abstract: The PhoPR two-component signal transduction system controls one of three responses activated by Bacillus subtilis to adapt to phosphate-limiting conditions (PHO response). The response involves the production of enzymes and transporters that scavenge for phosphate in the environment and assimilate it into the cell. However, in B. subtilis and some other Firmicutes bacteria, cell wall metabolism is also part of the PHO response due to the high phosphate content of the teichoic acids attached either to peptidoglycan (wall teichoic acid) or to the cytoplasmic membrane (lipoteichoic acid). Prompted by our observation that the phosphorylated WalR (WalR∼P) response regulator binds to more chromosomal loci than are revealed by transcriptome analysis, we established the PhoP∼P bindome in phosphate-limited cells. Here, we show that PhoP∼P binds to the chromosome at 25 loci: 12 are within the promoters of previously identified PhoPR regulon genes, while 13 are newly identified. We extend the role of PhoPR in cell wall metabolism showing that PhoP∼P binds to the promoters of four cell wall-associated operons (ggaAB, yqgS, wapA, and dacA), although none show PhoPR-dependent expression under the conditions of this study. We also show that positive autoregulation of phoPR expression and full induction of the PHO response upon phosphate limitation require PhoP∼P binding to the 3' end of the phoPR operon.
  
  
  OBJECTIVE
  The PhoPR two-component system controls one of three responses mounted by B. subtilis to adapt to phosphate limitation (PHO response). Here, establishment of the phosphorylated PhoP (PhoP∼P) bindome enhances our understanding of the PHO response in two important ways. First, PhoPR plays a more extensive role in adaptation to phosphate-limiting conditions than was deduced from transcriptome analyses. Among 13 newly identified binding sites, 4 are cell wall associated (ggaAB, yqgS, wapA, and dacA), revealing that PhoPR has an extended involvement in cell wall metabolism. Second, amplification of the PHO response must occur by a novel mechanism since positive autoregulation of phoPR expression requires PhoP∼P binding to the 3' end of the operon.
authors: Smurfit Institute of Genetics, Trinity College Dublin, Dublin, Ireland.
authors: Letal I
authors: LI
authors: Salzberg
authors: Smurfit Institute of Genetics, Trinity College Dublin, Dublin, Ireland.
authors: Eric
authors: E
authors: Botella
authors: Smurfit Institute of Genetics, Trinity College Dublin, Dublin, Ireland.
authors: Karsten
authors: K
authors: Hokamp
authors: Institute for Microbiology, Ernst Moritz Arndt University of Greifswald, Greifswald, Germany.
authors: Haike
authors: H
authors: Antelmann
authors: Institute for Microbiology, Ernst Moritz Arndt University of Greifswald, Greifswald, Germany.
authors: Sandra
authors: S
authors: Maaß
authors: Institute for Microbiology, Ernst Moritz Arndt University of Greifswald, Greifswald, Germany.
authors: Dörte
authors: D
authors: Becher
authors: Smurfit Institute of Genetics, Trinity College Dublin, Dublin, Ireland.
authors: David
authors: D
authors: Noone
authors: Smurfit Institute of Genetics, Trinity College Dublin, Dublin, Ireland kdevine@tcd.ie.
authors: Kevin M
authors: KM
authors: Devine
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002876:Chromosomes, Bacterial / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D016680:Genome, Bacterial; D008969:Molecular Sequence Data; D009876:Operon; D010710:Phosphates / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D010710:Phosphates; D012329:RNA, Bacterial; C061064:PhoP protein, Bacteria