PAS-LuxR transcriptional control of filipin biosynthesis in S. avermitilis - INSA Toulouse - Institut National des Sciences Appliquées de Toulouse Access content directly
Journal Articles Applied Microbiology and Biotechnology Year : 2014

PAS-LuxR transcriptional control of filipin biosynthesis in S. avermitilis

Cláudia M Vicente
Javier Santos-Aberturas
  • Function : Author
Tamara Payero
  • Function : Author
Eva Barreales
  • Function : Author
Antonio de Pedro
  • Function : Author
Jesús Aparicio

Abstract

The DNA region encoding the filipin gene cluster in Streptomyces avermitilis (pte) contains a PAS-LuxR regulatory gene, pteF, orthologue to pimM, the final pathway specific positive regulatory protein of pimaricin biosynthesis in Streptomyces natalensis. Gene replacement of the gene from S. avermitilis chromosome resulted in a severe loss of filipin production and delayed spore formation in comparison to that of the wild-type strain, suggesting that it acts as a positive regulator of filipin biosynthesis and that it may also have a role in sporulation. Complementation of the mutant with a single copy of the gene integrated into the chromosome restored wild-type phenotypes. Heterologous complementation with the regulatory counterpart from S. natalensis also restored parental phenotypes. Gene expression analyses in S. avermitilis wild-type and the mutant by reverse transcription-quantitative polymerase chain reaction of the filipin gene cluster suggested the targets for the regulatory protein. Transcription start points of all the genes of the cluster were studied by 5′-rapid amplification of complementary DNA ends. Transcription start point analysis of the pteF gene revealed that the annotated sequence in the databases is incorrect. Confirmation of target promoters was performed by in silico search of binding sites among identified promoters and the binding of the orthologous regulator for pimaricin biosynthesis PimM to gene promoters by electrophoretic mobility shift assays. Precise binding regions were investigated by DNAse I protection studies. Our results indicate that PteF activates the transcription from two promoters of polyketide synthase genes directly, and indirectly of other genes of the cluster.
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Dates and versions

hal-02559677 , version 1 (30-04-2020)

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Cláudia M Vicente, Javier Santos-Aberturas, Tamara Payero, Eva Barreales, Antonio de Pedro, et al.. PAS-LuxR transcriptional control of filipin biosynthesis in S. avermitilis. Applied Microbiology and Biotechnology, 2014, 98 (22), pp.9311-9324. ⟨10.1007/s00253-014-5998-7⟩. ⟨hal-02559677⟩
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