title: Expression of the Azotobacter vinelandii poly-beta-hydroxybutyrate biosynthetic phbBAC operon is driven by two overlapping promoters and is dependent on the transcriptional activator PhbR.
abstract: The Azotobacter vinelandii phbBAC genes encode the enzymes for poly-beta-hydroxybutyrate (PHB) synthesis. The phbR gene, which is located upstream of and in the opposite direction of phbBAC, encodes PhbR, a transcriptional activator which is a member of the AraC family of activators. Here we report that a mutation in phbR reduced PHB accumulation and transcription of a phbB-lacZ fusion. We also report that phbB is transcribed from two overlapping promoters, p(B)1 and p(B)2. The region corresponding to the -35 region of p(B)1 overlaps the p(B)2 -10 region. In the phbR mutant, expression of phbB from the p(B)1 promoter is significantly reduced, whereas expression from the p(B)2 promoter is slightly increased. Two phbR promoters, p(R)1 and p(R)2, were also identified. Transcription from p(R)2 was shown to be dependent on sigma(S). Six conserved 18-bp sites, designated R1 to R6, are present within the phbR-phbB intergenic region and are proposed to be putative binding targets for PhbR. R1 overlaps the -35 region of the p(B)1 promoter. A model for the regulation of phbB transcription by PhbR is proposed.
authors.affiliation: Departamento de Microbiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca Morelos 62250, Mexico.
authors.forename: Martín
authors.initials: M
authors.lastname: Peralta-Gil
authors.forename: Daniel
authors.initials: D
authors.lastname: Segura
authors.forename: Josefina
authors.initials: J
authors.lastname: Guzmán
authors.forename: Luis
authors.initials: L
authors.lastname: Servín-González
authors.forename: Guadalupe
authors.initials: G
authors.lastname: Espín
mesh_terms: D000102:Acetyl-CoA C-Acyltransferase / Q000235:genetics / Q000378:metabolism; D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism; D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism; D016948:Azotobacter vinelandii / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis; D021901:DNA, Intergenic / Q000032:analysis; D015964:Gene Expression Regulation, Bacterial; D006885:Hydroxybutyrates / Q000378:metabolism*; D008969:Molecular Sequence Data; D009876:Operon; D011091:Polyesters / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D000094122:Polyhydroxybutyrates
chemical_list: D000102:Acetyl-CoA C-Acyltransferase; D000217:Acyltransferases; D000429:Alcohol Oxidoreductases; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D021901:DNA, Intergenic; D006885:Hydroxybutyrates; D011091:Polyesters; D015534:Trans-Activators; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C023744:acetoacetyl-CoA reductase; C060970:poly-beta-hydroxybutyrate polymerase