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Characterization of a novel two-component regulatory system involved in the regulation of both actinorhodin and a type I polyketide in Streptomyces coelicolor

  • Applied Genetics and Molecular Biotechnology
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Abstract

To seek more information on function of two-component regulatory systems (TCSs) in Streptomycescoelicolor, a dozen TCS-knockout mutants were generated, and phenotype changes were determined. One TCS (SCO5403/5404)-deleted mutant with phenotype change was obtained. Here, we report the characterization of this novel TCS, designated as RapA1/A2 (regulation of both actinorhodin and a type I polyketide), using genetic and proteomic approaches. Although growth and morphological analyses showed no difference between the knockout mutant and wild-type strain M145, a visible decrease of the production of actinorhodin (Act) was observed in rapA1/A2 mutant. The decrease can be restored by introducing rapA1/A2 genes on an integrative vector. A 2D-gel based proteomic analysis showed that knockout of rapA1/A2 resulted in reduced expression of a putative 3-oxoacyl-[acyl-carrier protein] reductase that is part of a biosynthetic cluster for a cryptic type I polyketide. Further reverse-transcriptase-polymerase chain reaction (RT-PCR) analyses confirmed that expression levels of several biosynthetic genes and the respective pathway-specific regulatory genes actII-ORF4 and kasO for these two clusters were all down-regulated in the rapA1/A2 mutant, compared to M145. Taken together, the results demonstrated that RapA1/A2 may serve as a positive regulator for biosynthesis of both Act and the uncharacterized polyketide in S. coelicolor, and the effects exerted by RapA1/A2 were dependent on the pathway-specific regulatory genes.

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Acknowledgements

We are grateful to the Research Center for Proteome Analysis, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences for helps with 2D-PAGE and MOLDI-TOF analyses, and to Professor Keith Chater for kindly providing S. coelicolor cosmid 8F4, and all of the plasmids and strains used in PCR-targeting system.

This work was supported by National Natural Science Foundation of China (30470029), National Basic Research Program of China (2007CB707803) and the Knowledge Innovation Program of the Chinese Academy of Sciences (KSCX2-YW-G-007).

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Authors and Affiliations

  1. Laboratory of Molecular Microbiology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, People’s Republic of China

    Yinhua Lu, Weihua Wang, Dan Shu, Lei Chen, Zhongjun Qin, Sheng Yang & Weihong Jiang

  2. Center of Ecogenomics of the Biodesign Institute, Arizona State University, Tempe, AZ, 85287-6501, USA

    Weiwen Zhang

  3. Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, 200025, People’s Republic of China

    Weihong Jiang

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  1. Yinhua Lu
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  2. Weihua Wang
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  3. Dan Shu
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  4. Weiwen Zhang
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  5. Lei Chen
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Correspondence to Weihong Jiang.

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Lu, Y., Wang, W., Shu, D. et al. Characterization of a novel two-component regulatory system involved in the regulation of both actinorhodin and a type I polyketide in Streptomyces coelicolor . Appl Microbiol Biotechnol 77, 625–635 (2007). https://doi.org/10.1007/s00253-007-1184-5

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