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Volume 151, Issue 6

NovG, a DNA-binding protein acting as a positive regulator of novobiocin biosynthesis Free

Abstract

The biosynthetic gene cluster of the aminocoumarin antibiotic novobiocin contains two putative regulatory genes, i.e. and . The predicted gene product of shows a putative helix–turn–helix DNA-binding motif and shares sequence similarity with StrR, a well-studied pathway-specific transcriptional activator of streptomycin biosynthesis. Here functional proof is provided, by genetic and biochemical approaches, for the role of NovG as a positive regulator of novobiocin biosynthesis. The entire novobiocin cluster of the producer organism was expressed in the heterologous host M512, and additional strains were produced which lacked the gene within the heterologously expressed cluster. These Δ strains produced only 2 % of the novobiocin formed by the M512 strains carrying the intact novobiocin cluster. The production could be restored by introducing an intact copy of into the mutant. The presence of on a multicopy plasmid in the strain containing the intact cluster led to almost threefold overproduction of the antibiotic, suggesting that novobiocin biosynthesis is limited by the availability of NovG protein. Furthermore, purified N-terminal His-tagged NovG showed specific DNA-binding activity for the – and the – intergenic regions of the novobiocin and clorobiocin biosynthetic gene clusters, respectively. By comparing the DNA sequences of the fragments binding NovG, conserved inverted repeats were identified in both fragments, similar to those identified as the binding sites for StrR. The consensus sequence for the StrR and the putative NovG binding sites was GTTCRACTG(N)CRGTYGAAC. Therefore, NovG and StrR apparently belong to the same family of DNA-binding regulatory proteins.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): DIG, HTH, helix–turn–helix , PHD, Profile network prediction Heidelberg and SARP, Streptomyces antibiotic regulatory protein
SGM
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2005-06-01
2024-04-27
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NovG, a DNA-binding protein acting as a positive regulator of novobiocin biosynthesis
Microbiology 151, 1949 (2005); https://doi.org/10.1099/mic.0.27669-0
/content/journal/micro/10.1099/mic.0.27669-0
/content/journal/micro/10.1099/mic.0.27669-0
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