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The pathway-specific regulator AveR from Streptomyces avermitilis positively regulates avermectin production while it negatively affects oligomycin biosynthesis

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Abstract

The function of the regulatory protein AveR in Streptomyces avermitilis was examined. An aveR deletion mutant abolished avermectin production and produced more oligomycin, and its phenotype was complemented by a single copy of the aveR gene. Removal of the C-terminal HTH domain of AveR abolished avermectin biosynthesis, indicating the importance of HTH domain for AveR function. Promoter titration and promoter probe assays suggested that the transcription of aveA1, encoding polypeptide AVES1 of avermectin PKS, was activated by AveR. Chromatin immunoprecipitation (ChIP) assay showed that the predicted promoter regions of both the ave cluster and the olm cluster were target sites of AveR, and the DNA-binding activity of AveR was dependent on its HTH domain. RT-PCR analysis revealed that the transcriptions of ave structural genes were dependent on AveR, but that of olm structural genes and putative pathway-specific regulatory genes increased in the aveR mutants. Consistent with these observations, overexpression of aveR successfully increased avermectin production. These results indicated that aveR encodes a pathway-specific activator essential for avermectin biosynthesis and it also negatively affects oligomycin biosynthesis.

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References

  • Bibb MJ (2005) Regulation of secondary metabolism in streptomycetes. Curr Opin Microbiol 8:208–215

    Article  PubMed  CAS  Google Scholar 

  • Bierman M, Logan R, O’Brien K, Seno ET, Rao RN, Schoner BE (1992) Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene 116:43–49

    Article  PubMed  CAS  Google Scholar 

  • Burg RW, Miller BM, Baker EE, Birnbaum J, Currie SA, Hartman R, Kong YL, Monaghan RL, Olson G, Putter I, Tunac JB, Wallick H, Stapley EO, Oiwa R, Omura S (1979) Avermectins, new family of potent anthelmintic agents: producing organisms and fermentation. Antimicrob Agents Chemother 15:361–367

    PubMed  CAS  Google Scholar 

  • Chen Z, Wen J, Song Y, Wen Y, Li JL (2007) Enhancement and selective production of avermectin B by recombinants of Streptomyces avermitilis via intraspecific protoplast fusion. Chin Sci Bull 52:616–622

    Article  CAS  Google Scholar 

  • Chen L, Lu Y, Chen J, Zhang W, Shu D, Qin Z, Yang S, Jiang W (2008) Characterization of a negative regulator AveI for avermectin biosynthesis in Streptomyces avermitilis NRRL8165. Appl Microbiol Biotechnol 80:277–286

    Article  PubMed  CAS  Google Scholar 

  • Grainger DC, Overton TW, Reppas N, Wade JT, Tamai E, Hobman JL, Constantinidou C, Struhl K, Church G, Busby SJ (2004) Genomic Studies with Escherichia coli MelR protein: applications of chromatin immunoprecipitation and microarrays. J Bacteriol 186:6938–6943

    Article  PubMed  CAS  Google Scholar 

  • Hwang YS, Kim ES, Biro S, Choi CY (2003) Cloning and analysis of a DNA fragment stimulating avermectin production in various Streptomyces avermitilis strains. Appl Environ Microbiol 69:1263–1269

    Article  PubMed  CAS  Google Scholar 

  • Ikeda H, Omura S (1997) Avermectin biosynthesis. Chem Rev 97:2591–2610

    Article  PubMed  CAS  Google Scholar 

  • Ikeda H, Kotaki H, Tanaka H, Omura S (1988) Involvement of glucose catabolism in avermectin production by Streptomyces avermitilis. Antimicrob Agents Chemother 32:282–284

    PubMed  CAS  Google Scholar 

  • Ikeda H, Takada Y, Pang CH, Tanaka H, Omura S (1993) Transposon mutagenesis by Tn4560 and applications with avermectin-producing Streptomyces avermitilis. J Bacteriol 175:2077–2082

    PubMed  CAS  Google Scholar 

  • Ikeda H, Nonomiya T, Usami M, Ohta T, Omura S (1999) Organization of the biosynthetic gene cluster for the polyketide anthelmintic macrolide avermectin in Streptomyces avermitilis. Proc Natl Acad Sci USA 96:9509–9514

    Article  PubMed  CAS  Google Scholar 

  • Ikeda H, Nonomiya T, Omura S (2001) Organization of biosynthetic gene cluster for avermectin in Streptomyces avermitilis: analysis of enzymatic domains in four polyketide synthases. J Ind Microbiol Biotechnol 27:170–176

    Article  PubMed  CAS  Google Scholar 

  • Ikeda H, Ishikawa J, Hanamoto A, Shinose M, Kikuchi H, Shiba T, Sakaki Y, Hattori M, Omura S (2003) Complete genome sequence and comparative analysis of the industrial microorganism Streptomyces avermitilis. Nat Biotechnol 21:526–531

    Article  PubMed  Google Scholar 

  • Janssen GR, Bibb MJ (1993) Derivatives of pUC18 that have BglII sites flanking a multiple cloning site and that retain ability to identify recombinant clones by visual screening of Escherichia coli colonies. Gene 124:133–134

    Article  PubMed  CAS  Google Scholar 

  • Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA (2000) Practical Streptomyces genetics. The John Innes Foundation, Norwich

  • Kitani S, Ikeda H, Sakamoto T, Noguchi S, Nihira T (2009) Characterization of a regulatory gene, aveR, for the biosynthesis of avermectin in Streptomyces avermitilis. Appl Microbiol Biotechnol 82:1089–1096

    Article  PubMed  CAS  Google Scholar 

  • MacNeil DJ, Klapko LM (1987) Transformation of Streptomyces avermitilis by plasmid DNA. J Ind Microbiol 2:209–218

    Article  CAS  Google Scholar 

  • Omura S, Ikeda H, Ishikawa J, Hanamoto A, Takahashi C, Shinose M, Takahashi Y, Horikawa H, Nakazawa H, Osonoe T, Kikuchi H, Shiba T, Sakaki Y, Hattori M (2001) Genome sequence of an industrial microorganism Streptomyces avermitilis: deducing the ability of producing secondary metabolites. Proc Natl Acad Sci USA 98:12215–12220

    Article  PubMed  CAS  Google Scholar 

  • Pinna LA, Lorini M, Moret V, Siliprandi N (1967) Effect of oligomycin and succinate on mitochondrial metabolism of adenine nucleotides. Biochim Biophys Acta 143:18–25

    Article  PubMed  CAS  Google Scholar 

  • Rajkarnikar A, Kwon HJ, Ryu YW, Suh JW (2006) Catalytic domain of AfsKav modulates both secondary metabolism and morphologic differentiation in Streptomyces avermitilis ATCC 31272. Curr Microbiol 53:204–208

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

  • Stutzman-Engwall KJ, Price BS (2001) Streptomyces avermitilis regulatory genes for increased avermectin production. US Patent 6,197,591

  • Wilson DJ, Xue Y, Reynolds KA, Sherman DH (2001) Characterization and analysis of the PikD regulatory factor in the pikromycin biosynthetic pathway of Streptomyces venezuelae. J Bacteriol 183:3468–3475

    Article  PubMed  CAS  Google Scholar 

  • Zhao JL, Wen Y, Chen Z, Song Y, Li JL (2007) An adpA homologue in Streptomyces avermitilis is involved in regulation of morphogenesis and melanogenesis. Chin Sci Bull 52:623–630

    Article  CAS  Google Scholar 

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Acknowledgments

We thank Prof. Gang Liu (Chinese Academy of Science) and Prof. Linquan Bai (Shanghai Jiaotong University, China) for their critical reading of the manuscript. This work was supported by grants from the National Basic Research Program of China (Grant No. 2009CB118905) and the National High Technology Research and Development Program (Grant No. 2006AA10A209).

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Authors and Affiliations

  1. State Key Laboratory for Agro-biotechnology, College of Biological Sciences, China Agricultural University, 100193, Beijing, China

    Jia Guo, Jinlei Zhao, Lili Li, Zhi Chen, Ying Wen & Jilun Li

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  1. Jia Guo
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  2. Jinlei Zhao
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  3. Lili Li
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  4. Zhi Chen
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  5. Ying Wen
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  6. Jilun Li
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Corresponding author

Correspondence to Ying Wen.

Additional information

Communicated by G. Klug.

J. Guo and J. Zhao contributed equally to this work.

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Guo, J., Zhao, J., Li, L. et al. The pathway-specific regulator AveR from Streptomyces avermitilis positively regulates avermectin production while it negatively affects oligomycin biosynthesis. Mol Genet Genomics 283, 123–133 (2010). https://doi.org/10.1007/s00438-009-0502-2

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