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

Eustáquio, Alessandra S.; Li, Shu-Ming; Heide, Lutz

doi:10.1099/mic.0.27669-0

Cited by 35 publications

(21 citation statements)

References 52 publications

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“…Functional studies of StrR like regulators have been limited to StrR [Beyer et al, 1996;Retzlaff and Distler, 1995], KasT from the kasugamycin producer Streptomyces kasugaensis [Ikeno et al, 2002] and NovG from the novobiocin producer Streptomyces spheroides [Eustaquio et al, 2005]. Our DNA-binding studies with Bbr provide the first characterization of a regulator of biosynthesis genes for glycopeptide antibiotics.…”

Section: Bbr Binds Directly To the Promoter Region Of The Balhimycin mentioning

confidence: 99%

The Border Sequence of the Balhimycin Biosynthesis Gene Cluster from Amycolatopsis balhimycina Contains bbr, Encoding a StrR-Like Pathway-Specific Regulator

et al. 2007

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Balhimycin, produced by the actinomycete Amycolatopsis balhimycina DSM5908, is a glycopeptide antibiotic highly similar to vancomycin, the antibiotic of ‘last resort’ used for the treatment of resistant Gram-positive pathogenic bacteria. Partial sequence of the balhimycin biosynthesis gene cluster was previously reported. In this work, cosmids which overlap the region of the characterized gene cluster were isolated and sequenced. At the ‘left’ end of the cluster, genes were identified which are involved in balhimycin biosynthesis, transport, resistance and regulation. The ‘right’ end border is defined by a putative 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase (dahp) gene. The proximate gene is similar to a type I polyketide synthase gene of the rifamycin producer Amycolatopsis mediterranei indicating that another biosynthesis gene cluster might be located directly next to the balhimycin gene cluster. The newly identified StrR-like pathway-specific regulator, Bbr, was characterized to be a DNA-binding protein and may have a role in balhimycin biosynthesis. Purified N-terminally His-tagged Bbr shows specific DNA-binding to five promoter regions within the gene cluster. By in silico analysis and by comparison of the DNA sequences binding Bbr, conserved inverted repeat sequences for the Bbr-binding site are proposed. The putative Bbr consensus sequence differs from that published for StrR.

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Section: Bbr Binds Directly To the Promoter Region Of The Balhimycin mentioning

confidence: 99%

The Border Sequence of the Balhimycin Biosynthesis Gene Cluster from Amycolatopsis balhimycina Contains bbr, Encoding a StrR-Like Pathway-Specific Regulator

et al. 2007

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“…StrR is a well-studied pathway-specific transcriptional regulator that activates the expression of streptomycin biosynthetic genes in Streptomyces griseus and Streptomyces glaucescens (28). Other StrR-like regulators have been characterized, such as NovG from the Streptomyces caeruleus novobiocin cluster (10), CloG from the Streptomyces roseochromogenes clorobiocin cluster (10), and KasT from the Streptomyces kasugaensis kasugamycin cluster (13). Recently, the production of A40926 was found to be influenced by phosphate (12,42).…”

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confidence: 99%

Phosphate-Controlled Regulator for the Biosynthesis of the Dalbavancin Precursor A40926

et al. 2007

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The actinomycete Nonomuraea sp. strain ATCC 39727 produces the glycopeptide A40926, the precursor of the novel antibiotic dalbavancin. Previous studies have shown that phosphate limitation results in enhanced A40926 production. The A40926 biosynthetic gene (dbv) cluster, which consists of 37 genes, encodes two putative regulators, Dbv3 and Dbv4, as well as the response regulator (Dbv6) and the sensor-kinase (Dbv22) of a putative two-component system. Reverse transcription-PCR (RT-PCR) and real-time RT-PCR analysis revealed that the dbv14-dbv8 and the dbv30-dbv35 operons, as well as dbv4, were negatively influenced by phosphate. Dbv4 shows a putative helix-turn-helix DNA-binding motif and shares sequence similarity with StrR, the transcriptional activator of streptomycin biosynthesis in Streptomyces griseus. Dbv4 was expressed in Escherichia coli as an N-terminal His 6 -tagged protein. The purified protein bound the dbv14 and dbv30 upstream regions but not the region preceding dbv4. Bbr, a Dbv4 ortholog from the gene cluster for the synthesis of the glycopeptide balhimycin, also bound to the dbv14 and dbv30 upstream regions, while Dbv4 bound appropriate regions from the balhimycin cluster. Our results provide new insights into the regulation of glycopeptide antibiotics, indicating that the phosphate-controlled regulator Dbv4 governs two key steps in A40926 biosynthesis: the biosynthesis of the nonproteinogenic amino acid 3,5-dihydroxyphenylglycine and critical tailoring reactions on the heptapeptide backbone.

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“…These observations are not unique to the SARP family. NovG, a member of the StrR family, has been characterized as a transcriptional activator for the novobiocin gene cluster (12). PikD, a member of the LAL family, has been characterized as a transcriptional activator for the pikromycin gene cluster (55), and most recently ThnU, a member of the Lys-R family, was found to be essential for the transcription of genes essential for thienamycin biosynthesis in S. cattleya (39).…”

Section: Discussionmentioning

confidence: 99%

“…In Streptomyces coelicolor A3(2), the pleotropic regulator AfsR has been characterized and found to control the production of actinorhodin, undecylprodigiosin, and calcium-dependent antibiotic (CDA). There are many examples of CSRs that have been characterized and shown to regulate transcription of genes of a single secondary metabolite biosynthetic cluster (12). These late-stage regulators, usually transcriptional activators, are represented by several families, including the Streptomyces antibiotic regulatory protein (SARP) family characterized by an N-terminal OmpR-like domain, the StrR family characterized by a helix-turn-helix motif in its central region, and the LAL family characterized by a LuxR-like C-terminal DNA binding motif (8).…”

mentioning

confidence: 99%

Identification and Characterization of NocR as a Positive Transcriptional Regulator of the β-Lactam Nocardicin A inNocardia uniformis

Davidsen

Townsend

2009

J Bacteriol

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Nocardicin A is a monocyclic ␤-lactam isolated from the actinomycete Nocardia uniformis, which shows moderate activity against a broad spectrum of gram-negative bacteria. Within the biosynthetic gene cluster of nocardicin A, nocR encodes a 583-amino-acid protein with high similarity to a class of transcriptional regulators known as streptomyces antibiotic regulatory proteins. Insertional inactivation of this gene resulted in a mutant showing morphology and growth characteristics similar to the wild type, but one that did not produce detectable levels of nocardicin A or the early precursor p-hydroxybenzoyl formate. Similar disruptions of nocD, nocE, and nocO yielded mutants that maintained production of nocardicin A at levels similar to the wild-type strain. In trans complementation of the nocR::apr mutant partially restored the wild-type phenotype. Transcriptional analysis of the nocR::apr mutant using reverse transcription-PCR found an absence of mRNA transcripts for the early-stage nocardicin A biosynthetic genes. In addition, transcription of the genes responsible for the biosynthesis of the nonproteinogenic p-hydroxyphenylglycine (pHPG) precursor was attenuated on the nocR disruption mutant. NocR was heterologously expressed and purified from Escherichia coli as an N-terminal maltose binding protein-tagged fusion protein. DNA binding assays demonstrated that NocR is a DNA binding protein, targeting the 126-bp intergenic region between nocF and nocA. Within this intergenic region is the likely binding motif, a direct hexameric repeat, TGATAA, with a 5-bp spacer. These experiments establish NocR as a positive transcriptional regulator of the nocardicin A biosynthetic pathway, coordinating the initial steps of nocardicin A biosynthesis to the production of its pHPG precursor.

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

Cited by 35 publications

References 52 publications

The Border Sequence of the Balhimycin Biosynthesis Gene Cluster from <i>Amycolatopsis balhimycina</i> Contains <i>bbr</i>, Encoding a StrR-Like Pathway-Specific Regulator

The Border Sequence of the Balhimycin Biosynthesis Gene Cluster from <i>Amycolatopsis balhimycina</i> Contains <i>bbr</i>, Encoding a StrR-Like Pathway-Specific Regulator

Phosphate-Controlled Regulator for the Biosynthesis of the Dalbavancin Precursor A40926

Identification and Characterization of NocR as a Positive Transcriptional Regulator of the β-Lactam Nocardicin A inNocardia uniformis

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