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From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes Tü6028
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From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes Tü6028

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PimM, a PAS domain positive regulator of pimaricin biosynthesis in Streptomyces natalensis

Nuria Antón1,2, Javier Santos-Aberturas1,2, Marta V Mendes2

  • 1Area of Microbiology, Faculty of Biology, University of León, 24071 León, Spain.

Microbiology (Reading, England)
|September 5, 2007
PubMed

Insights

PimM is a novel positive regulator of pimaricin biosynthesis in Streptomyces natalensis. Deletion of pimM halts pimaricin production, while its presence enhances it, revealing key regulatory targets.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • Pimaricin is an antifungal polyketide produced by Streptomyces natalensis.
  • The regulation of secondary metabolite biosynthesis in bacteria is complex and often involves pathway-specific regulators.
  • Understanding these regulatory mechanisms is crucial for optimizing antibiotic production.

Purpose of the Study:

  • To identify and characterize novel regulatory elements involved in pimaricin biosynthesis.
  • To elucidate the role of the newly identified gene, pimM, in the pimaricin gene cluster.
  • To investigate the regulatory mechanism of PimM on pimaricin production.

Main Methods:

  • DNA sequencing and gene cloning to identify pimM.
  • Gene replacement and complementation studies to assess pimM function.
  • Reverse transcriptase PCR (RT-PCR) to analyze gene expression of the pimaricin cluster.
  • Bioinformatic analysis to predict protein domains and homologies.

Main Results:

  • A novel gene, pimM, was identified within the pimaricin gene cluster, encoding a putative regulatory protein.
  • PimM possesses an N-terminal PAS domain and a C-terminal helix-turn-helix (HTH) DNA-binding motif.
  • Deletion of pimM abolished pimaricin production, confirming its essential role as a positive regulator.
  • Overexpression of pimM in wild-type S. natalensis significantly increased pimaricin production.
  • RT-PCR analysis revealed that PimM regulates genes involved in the initiation and early elongation of polyketide synthesis.
  • PimM appears to function independently of the previously identified regulator, PimR.

Conclusions:

  • PimM is a critical positive regulator for pimaricin biosynthesis in Streptomyces natalensis.
  • The HTH motif in PimM is essential for its DNA-binding and regulatory function.
  • PimM controls early steps in polyketide chain assembly, a key regulatory point.
  • PimM represents a distinct regulatory pathway, independent of PimR, highlighting the intricate control of pimaricin production.

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