Transcriptional control by A-factor of strR, the pathway-specific transcriptional activator for streptomycin biosynthesis in Streptomyces griseus.
A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone) triggers streptomycin production by inducing the transcription of strR, encoding the pathway-specific transcriptional activator, through signal transduction in the A-factor regulatory cascade in Streptomyces griseus. AdpA, one of the key transcriptional activators in the cascade, bound two upstream activation sites, approximately at nucleotide positions -270 and -50 with respect to the transcriptional start point of strR, as determined by gel mobility shift assays and DNase I footprinting. Transcriptional analysis of the strR promoter with mutated AdpA-binding sites showed that both sites were required for full transcriptional activation of strR by AdpA. Potassium permanganate footprinting showed that AdpA assisted RNA polymerase in forming an open complex at an appropriate position for transcriptional initiation of strR. Nine transcriptional units within the streptomycin biosynthesis gene cluster, including the strR-aphD operon, depended on StrR, indicating that StrR is the pathway-specific transcriptional activator for the whole gene cluster. Consistent with this, expression of strR under the control of a constitutively expressed promoter in an adpA null mutant caused the host to produce streptomycin.
Tomono
Ayami
A
Department of Biotechnology, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan.
Tsai
Yisan
Y
Yamazaki
Haruka
H
Ohnishi
Yasuo
Y
Horinouchi
Sueharu
S
D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D003850:Deoxyribonuclease I; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D005810:Multigene Family; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000502:physiology; D013305:Streptomyces griseus / Q000235:genetics* / Q000378:metabolism*; D013307:Streptomycin / Q000096:biosynthesis; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic / Q000502:physiology
C401645:AdpA protein, Streptomyces griseus; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C098255:StrR protein, Streptomyces griseus; D015534:Trans-Activators; D014157:Transcription Factors; C025035:A-factor (Streptomyces); D003850:Deoxyribonuclease I; D015107:4-Butyrolactone; D013307:Streptomycin