Involvement of AlpV, a New Member of the
 Streptomyces
 Antibiotic Regulatory Protein Family, in Regulation of the Duplicated Type II Polyketide Synthase
 alp
 Gene Cluster in
 Streptomyces ambofaciens

Aigle, Bertrand; Pang, Xiuhua; Decaris, Bernard; Leblond, Pierre

doi:10.1128/jb.187.7.2491-2500.2005

Cited by 36 publications

(33 citation statements)

References 46 publications

(64 reference statements)

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“…4). In the parent strain, the expression of the alpA biosynthetic gene was induced during the late transition phase and persisted until entry into the stationary phase, as previously observed (1,8,41). In the alpW mutant, alpA transcripts were also detected from the transition phase, but in contrast to the wild type, the transcripts were detected 1 h earlier and were strongly maintained through the stationary phase.…”

Section: Vol 193 2011 Regulation Of Kinamycin Biosynthesis In S Amsupporting

confidence: 71%

“…Streptomyces strains were manipulated as described previously (25,41). Pigment and antibiotic production were assessed on or in R2 medium as described previously (1,41). Escherichia coli strains were routinely cultivated in Luria-Bertani (LB) and SOB liquid media (44).…”

Section: Methodsmentioning

confidence: 99%

“…The product of the alpV gene was previously shown to be essential for the production of both the orange pigment and alpomycin by activating the transcription of the structural biosynthetic genes (1). The transcription of alpV is itself under the control of the product of alpZ, a ␥-butyrolactone (GBL)-like receptor homologue from the TetR superfamily of transcription factors (for a review, see reference 43), which acts at the top of the regulatory cascade controlling the activation/repression of the alp pathway (8).…”

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confidence: 99%

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Characterization and Manipulation of the Pathway-Specific Late Regulator AlpW RevealsStreptomyces ambofaciensas a New Producer of Kinamycins

et al. 2011

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The genome sequence of Streptomyces ambofaciens, a species known to produce the congocidine and spiramycin antibiotics, has revealed the presence of numerous gene clusters predicted to be involved in the biosynthesis of secondary metabolites. Among them, the type II polyketide synthase-encoding alp cluster was shown to be responsible for the biosynthesis of a compound with antibacterial activity. Here, by means of a deregulation approach, we gained access to workable amounts of the antibiotics for structure elucidation. These compounds, previously designated as alpomycin, were shown to be known members of kinamycin family of antibiotics. Indeed, a mutant lacking AlpW, a member of the TetR regulator family, was shown to constitutively produce kinamycins. Comparative transcriptional analyses showed that expression of alpV, the essential regulator gene required for activation of the biosynthetic genes, is strongly maintained during the stationary growth phase in the alpW mutant, a stage at which alpV transcripts and thereby transcripts of the biosynthetic genes normally drop off. Recombinant AlpW displayed DNA binding activity toward specific motifs in the promoter region of its own gene and that of alpV and alpZ. These recognition sequences are also targets for AlpZ, the ␥-butyrolactone-like receptor involved in the regulation of the alp cluster. However, unlike that of AlpZ, the AlpW DNA-binding ability seemed to be insensitive to the signaling molecules controlling antibiotic biosynthesis. Together, the results presented in this study reveal S. ambofaciens to be a new producer of kinamycins and AlpW to be a key late repressor of the cellular control of kinamycin biosynthesis.Streptomycetes are filamentous, soil-dwelling bacteria that undergo a complex morphological differentiation correlated with a rich biochemical specialization occurring during the late stages of growth. These prokaryotes are known for their capacity to biosynthesize a vast array of important secondary metabolites used in human activities, including antibiotics, antitumor agents, immunosuppressants, antihelmenthics, and herbicides. In spite of decades of genetic studies and industrial uses, members of the genus Streptomyces have recently revealed, by means of genomic analyses, their hitherto unsuspected ability to produce further novel secondary metabolites with potentially useful activities (5, 22, 37).In Streptomyces ambofaciens ATCC 23877, which was previously known to produce only the antibiotics congocidine (12) and spiramycin (42), the sequencing of the terminal regions of the linear chromosome (over circa 3 Mb; accession no. AM238663 and AM238664, respectively, for the left and right arms) has unveiled 14 novel secondary metabolite gene clusters (http://www.weblgm.scbiol.uhp-nancy.fr/ambofaciens/) (10). Among them, two clusters were experimentally shown to be involved in the biosynthesis of the siderophore coelichelin (3) and the pyrrole-amide congocidine (23), respectively. In our groups, the function of the duplicated type II polyketi...

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Section: Vol 193 2011 Regulation Of Kinamycin Biosynthesis In S Amsupporting

confidence: 71%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Characterization and Manipulation of the Pathway-Specific Late Regulator AlpW RevealsStreptomyces ambofaciensas a New Producer of Kinamycins

et al. 2011

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“…The regulatory effect of pathway-specific regulators are usually confined to genes in the same cluster. The best-studied pathway-specific regulators include ActII-ORF4 (Fernández-Moreno et al, 1991), RedD (Feitelson et al, 1985;Narva & Feitelson, 1990), DnrI (Stutzman-Engwall et al, 1992) and AlpV (Aigle et al, 2005). Multiple pathway-specific regulators often exist in a gene cluster, forming a regulatory cascade (Cundliffe, 2008;Pang et al, 2004).…”

Section: Discussionmentioning

confidence: 99%

Regulation of the biosynthesis of thiopeptide antibiotic cyclothiazomycin by the transcriptional regulator SHJG8833 in Streptomyces hygroscopicus 5008

Zhang

Chen

et al. 2014

Microbiology

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Cyclothiazomycin is a member of the thiopeptide antibiotics, which are usually complicated derivatives of ribosomally synthesized peptides. A gene cluster containing 12 ORFs identical to the clt cluster encoding cyclothiazomycin from Streptomyces hygroscopicus 10-22 was revealed by genome sequencing in S. hygroscopicus 5008. Genes SHJG8833 and SHJG8837 of the cluster and flanking gene SHJG8838 were predicted to encode regulatory proteins from different families. In this study, we showed that the newly identified cluster is functional and we investigated the roles of these regulatory genes in the regulation of cyclothiazomycin biosynthesis. We determined that SHJG8833, but not SHJG8837 or SHJG8838, is critical for cyclothiazomycin biosynthesis. The transcriptional start point of SHJG8833 was located to a thymidine 54 nt upstream of the start codon. Inactivation of SHJG8833 abrogated the production of cyclothiazomycin, and synthesis could be restored by reintroducing SHJG8833 into the mutant strain. Gene expression analyses indicated that SHJG8833 regulates a consecutive set of seven genes from SHJG8826 to SHJG8832, whose products are predicted to be involved in different steps in the construction of the main framework of cyclothiazomycin. Transcriptional analysis indicated that these seven genes may form two operons, SHJG8826-27 and SHJG8828-32. Gel-shift analysis demonstrated that the DNA-binding domain of SHJG8833 binds the promoters of SHJG8826 and SHJG8828 and sequences internal to SHJG8826 and SHJG8829, and a conserved binding sequence was deduced. These results indicate that SHJG8833 is a positive regulator that controls cyclothiazomycin biosynthesis by activating structural genes in the clt cluster.

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“…Among them, alpV was previously shown to be essential for the production of both the orange pigment and alpomycin (1). It has been postulated that the AlpV protein occupies the critical position in the alp regulatory cascade, leading to the activation of transcription of the biosynthetic genes, although direct binding to promoter regions within the cluster has not yet been demonstrated.…”

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confidence: 99%

Regulation of the Synthesis of the Angucyclinone Antibiotic Alpomycin inStreptomyces ambofaciensby the Autoregulator Receptor AlpZ and Its Specific Ligand

Bunet¹,

Mendes²,

Rouhier

et al. 2008

J Bacteriol

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Streptomyces ambofaciens produces an orange pigment and the antibiotic alpomycin, both of which are products of a type II polyketide synthase gene cluster identified in each of the terminal inverted repeats of the linear chromosome. Five regulatory genes encoding Streptomyces antibiotic regulatory proteins (alpV, previously shown to be an essential activator gene; alpT; and alpU) and TetR family receptors (alpZ and alpW) were detected in this cluster. Here, we demonstrate that AlpZ, which shows high similarity to ␥-butyrolactone receptors, is at the top of a pathway-specific regulatory hierarchy that prevents synthesis of the alp polyketide products. Deletion of the two copies of alpZ resulted in the precocious production of both alpomycin and the orange pigment, suggesting a repressor role for AlpZ. Consistent with this, expression of the five alp-located regulatory genes and of two representative biosynthetic structural genes (alpA and alpR) was induced earlier in the alpZ deletion strain. Furthermore, recombinant AlpZ was shown to bind to specific DNA sequences within the promoter regions of alpZ, alpV, and alpXW, suggesting direct transcriptional control of these genes by AlpZ. Analysis of solvent extracts of S. ambofaciens cultures identified the existence of a factor which induces precocious production of alpomycin and pigment in the wild-type strain and which can disrupt the binding of AlpZ to its DNA targets. This activity is reminiscent of ␥-butyrolactone-type molecules. However, the AlpZ-interacting molecule(s) was shown to be resistant to an alkali treatment capable of inactivating ␥-butyrolactones, suggesting that the AlpZ ligand(s) does not possess a lactone functional group.

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Involvement of AlpV, a New Member of the Streptomyces Antibiotic Regulatory Protein Family, in Regulation of the Duplicated Type II Polyketide Synthase alp Gene Cluster in Streptomyces ambofaciens

Cited by 36 publications

References 46 publications

Characterization and Manipulation of the Pathway-Specific Late Regulator AlpW RevealsStreptomyces ambofaciensas a New Producer of Kinamycins

Characterization and Manipulation of the Pathway-Specific Late Regulator AlpW RevealsStreptomyces ambofaciensas a New Producer of Kinamycins

Regulation of the biosynthesis of thiopeptide antibiotic cyclothiazomycin by the transcriptional regulator SHJG8833 in Streptomyces hygroscopicus 5008

Regulation of the Synthesis of the Angucyclinone Antibiotic Alpomycin inStreptomyces ambofaciensby the Autoregulator Receptor AlpZ and Its Specific Ligand

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