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Journal of Bacteriology, August 2005, p. 5595-5604, Vol. 187, No. 16
0021-9193/05/$08.00+0     doi:10.1128/JB.187.16.5595-5604.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Transcriptional Control by A-Factor of strR, the Pathway-Specific Transcriptional Activator for Streptomycin Biosynthesis in Streptomyces griseus

Ayami Tomono, Yisan Tsai, Haruka Yamazaki, Yasuo Ohnishi, and Sueharu Horinouchi^*

Department of Biotechnology, Graduate School of Agriculture and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan

Received 23 March 2005/ Accepted 24 May 2005

A-factor (2-isocapryloyl-3R-hydroxymethyl--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.

Present address: Division of Molecular Neurobiology, Institute of Medical Sciences, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan.

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