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A γ-butyrolactone autoregulator-receptor system involved in the regulation of auricin production in Streptomyces aureofaciens CCM 3239

  • Applied genetics and molecular biotechnology
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Abstract

The γ-butyrolactone (GBL) autoregulator–receptor systems play a role in controlling secondary metabolism and/or morphological differentiation in many Streptomyces species. We previously identified the aur1 gene cluster, located on the Streptomyces aureofaciens CCM 3239 large linear plasmid pSA3239, which is responsible for the production of the angucycline antibiotic auricin. Here, we describe the characterisation of two genes, sagA and sagR, encoding GBL autoregulatory signalling homologues, which lie in the upstream part of the aur1 cluster. SagA was similar to GBL synthases and SagR to GBL receptors. The expression of each gene is directed by its own promoter, sagAp for sagA and sagRp for sagR. Both genes were active mainly during the exponential phase, and their transcription was interdependent. The disruption of sagA abolished auricin production, while the disruption of sagR resulted in precocious but dramatically reduced auricin production. Transcription from the aur1Pp and aur1Rp promoters, which direct the expression of auricin-specific cluster-situated regulators (CSRs), was also precocious and increased in the sagR mutant strain. In addition, SagR was also shown to specifically bind both promoters in vitro. These results indicated that the SagA–SagR GBL system regulates auricin production. Unlike many other GBL receptors, SagR does not bind its own promoter, but Aur1R, an auricin-specific repressor from the family of pseudo GBL receptors, does bind both sagAp and sagRp promoters. Moreover, the expression of both promoters was deregulated in an aur1R mutant, indicating that the SagA–SagR GBL system is regulated by a feedback mechanism involving the auricin-specific CSR Aur1R, which regulates downstream.

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Acknowledgments

We are grateful to Dr. Mark Buttner (John Innes Centre, Norwich, UK) for the pPM929 plasmid and to Dr. Bertold Gust (John Innes Centre, Norwich, UK) for kindly providing all the plasmids and strains used in the PCR-targeting system; the system itself was supplied by Plant Bioscience Ltd. (Norwich, UK). This work was supported by the Slovak Research and Development Agency under contract No. APVV-0203-11. The research leading to these results received funding from the European Commission’s Seventh Framework Programme (FP7/2007-2013) under the grant agreement STREPSYNTH (project No. 613877).

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  1. Institute of Molecular Biology, Slovak Academy of Sciences, Dubravska Cesta 21, 845 51, Bratislava, Slovak Republic

    Erik Mingyar, Lubomira Feckova, Renata Novakova, Carmen Bekeova & Jan Kormanec

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  1. Erik Mingyar
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  2. Lubomira Feckova
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  3. Renata Novakova
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  4. Carmen Bekeova
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  5. Jan Kormanec
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Correspondence to Jan Kormanec.

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Mingyar, E., Feckova, L., Novakova, R. et al. A γ-butyrolactone autoregulator-receptor system involved in the regulation of auricin production in Streptomyces aureofaciens CCM 3239. Appl Microbiol Biotechnol 99, 309–325 (2015). https://doi.org/10.1007/s00253-014-6057-0

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