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Published online ahead of print on 21 April 2009 as doi:10.1099/mic.0.028639-0
Microbiology 2009;155:1819.

Microbiology (2009), DOI 10.1099/mic.0.028639-0
© 2009 Society for General Microbiology
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Microbiology 0 (2009), mic.0.028639; DOI  10.1099/mic.0.028639-0
© 2009 Society for General Microbiology

polR, a pathway-specific transcriptional regulatory gene, positively controls polyoxin biosynthesis in Streptomyces cacaoi subsp. asoensis

R. Li1, Z. Xie1, Y. Tian1, H. Yang1, W. Chen2, D. You2, G. Liu3, Z. Deng4 and H. Tan5,6

1 State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences;
2 Laboratory of Microbial Metabolism and School of Life Science & Biotechnology, Shanghai Jiaotong Un;
3 1State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences;
4 Laboratory of Microbial Metabolism and School of Life Science & Biotechnology, Shanghai Jiaotong Unv;
5 State Key Laboratory of Microbial Resources, Institute of Microbiology,Chinese Academy of Sciences

ABSTRACT

The polyoxin (POL) biosynthetic gene cluster (pol) was recently cloned from Streptomyces cacaoi var. asoensis. A 3.3-kb DNA fragment carrying an obvious open reading frame (polR), whose deduced product shows sequence similarity to SanG of Streptomyces ansochromogenes and PimR of Streptomyces natalensis, was revealed within the pol gene cluster. Disruption of polR abolished POL production, which could be complemented in turn by the integration of a single copy polR into the chromosome of the non-producing mutant. The introduction of an extra copy of polR in wild-type strain resulted in increase in the production of POLs. The transcription start point (tsp) of polR was determined by S1 mapping. Reverse transcriptase PCR (RT-PCR) experiment showed that PolR is required for the transcription of 18 structural genes in the pol gene cluster. Furthermore, we showed that polC and polB, the respective first gene of two putative operons consisting of the 18 structural genes, have similar promoter structures. Gel retardation assay indicated that PolR has specific DNA-binding activity for the promoter regions of polC and polB. Our data suggested that PolR acts in a positive manner to regulate the POL production by activating the transcription of at least two putative operons in the pol gene cluster.

6 E-mail: tanhr{at}sun.im.ac.cn






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