title: Autorepression of AdpA of the AraC/XylS family, a key transcriptional activator in the A-factor regulatory cascade in Streptomyces griseus.
abstract: AdpA belonging to the AraC/XylS family is a key transcriptional activator in the A-factor regulatory cascade in Streptomyces griseus, activating a number of genes required for physiological and morphological differentiation. On the other hand, AdpA repressed its own transcription by cooperative binding to the promoter region containing multiple operator sites. AdpA contained three operator sites, site 1 approximately at nucleotide position -100, site 2 at the promoter elements, and site 3 at position +80. AdpA bound to a strong binding site 1 increased the affinity for AdpA of a weak site 2, probably by forming a DNA loop via the two molecules of AdpA dimer, thus preventing RNA polymerase from access to the promoter. AdpA bound to site 3 with rather weak affinity repressed the AdpA promoter activity independently of sites 1 and 2, perhaps preventing RNA polymerase from chain elongation. Consistent with this model, the in vivo transcription of AdpA containing mutated site 1 or site 3 was greatly increased. In addition, streptomycin production, one of the phenotypes controlled positively by AdpA, was greatly increased in the mutants containing AdpA with a mutation at site 1 and site 3. The in vitro transcription of AdpA containing mutated site 1 was also increased. Thus, the transcription of AdpA, encoding an important transcriptional factor for ordered physiological and morphological development, is self-controlled.
authors.affiliation: Department of Biotechnology, Graduate School of Agriculture and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Jun-Ya
authors.initials: JY
authors.lastname: Kato
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Ohnishi
authors.forename: Sueharu
authors.initials: S
authors.lastname: Horinouchi
mesh_terms: D001426:Bacterial Proteins / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000145:classification / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009875:Operator Regions, Genetic; D011401:Promoter Regions, Genetic / Q000235:genetics; D012097:Repressor Proteins / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D016415:Sequence Alignment; D013305:Streptomyces griseus / Q000235:genetics*; D013307:Streptomycin / Q000096:biosynthesis; D015534:Trans-Activators / Q000145:classification / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism*
chemical_list: C401645:AdpA protein, Streptomyces griseus; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; D003850:Deoxyribonuclease I; D013307:Streptomycin