title: Intramolecular signal transduction within the FixJ transcriptional activator: in vitro evidence for the inhibitory effect of the phosphorylatable regulatory domain.
abstract: FixJ is a phosphorylatable 'response regulator' controlling the transcription of the key nitrogen fixation genes nifA and fixK in Rhizobium meliloti. Sequence and genetic analyses indicated that FixJ comprises an N-terminal phosphorylatable regulatory domain, FixJN, and a C-terminal transcriptional activator domain, FixJC. We have now overexpressed and purified the FixJC protein and show that it is fully active in an in vitro transcription system with purified RNA polymerase. FixJC appeared to act synergistically with RNA polymerase at the nifA promoter. Furthermore FixJC was more active in vitro than the full-length dephosphorylated FixJ protein. Therefore activity of FixJC is inhibited by FixJN within the FixJ protein. This inhibition is relieved by phosphorylation of FixJN. Such a negative mode of intramolecular signal transduction may be generalizable to other response regulators.
authors.affiliation: Laboratoire de Biologie Moléculaire des Relations Plantes-Microorganismes, CNRS-INRA, Castanet-Tolosan, France.
authors.forename: S
authors.initials: S
authors.lastname: Da Re
authors.forename: S
authors.initials: S
authors.lastname: Bertagnoli
authors.forename: J
authors.initials: J
authors.lastname: Fourment
authors.forename: J M
authors.initials: JM
authors.lastname: Reyrat
authors.forename: D
authors.initials: D
authors.lastname: Kahn
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D001665:Binding Sites; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D009586:Nitrogen Fixation / Q000235:genetics; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D015398:Signal Transduction; D014158:Transcription, Genetic
chemical_list: D001426:Bacterial Proteins; C069289:FixJ protein, Bacteria; D012321:DNA-Directed RNA Polymerases