The two-component phoR-phoP system of Streptomyces natalensis: Inactivation or deletion of phoP reduces the negative phosphate regulation of pimaricin biosynthesis

Mendes, Marta V.; Tunca, Sedef; Antón, Nuria; Recio, Eliseo; Sola‐Landa, Alberto; Aparicio, Jesús F.; Martı́n, Juan F.

doi:10.1016/j.ymben.2006.10.003

Cited by 94 publications

(80 citation statements)

References 40 publications

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“…Given that GST-tagged proteins have been successfully used in EMSAs (22,32), GST-PimM fusion proteins were used for in vitro experiments. Both the complete PimM protein and truncated forms of the protein lacking the PAS domain (PimM ⌬PAS ) and containing just the DNA-binding domain (PimM DBD ) were found to bind with high affinity to the promoter regions of the same pim genes and to produce the same pattern of shifted bands in each case, thus suggesting that binding specificity is independent of the PAS domain.…”

Section: Discussionmentioning

confidence: 99%

Molecular Control of Polyene Macrolide Biosynthesis

Santos‐Aberturas

Vicente

Guerra

et al. 2011

Journal of Biological Chemistry

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Control of polyene macrolide production in Streptomyces natalensis is mediated by the transcriptional activator PimM. This regulator, which combines an N-terminal PAS domain with a C-terminal helix-turn-helix motif, is highly conserved among polyene biosynthetic gene clusters. PimM, truncated forms of the protein without the PAS domain (PimM ⌬PAS ), and forms containing just the DNA-binding domain (DBD) (PimM DBD ) were overexpressed in Escherichia coli as GSTfused proteins. GST-PimM binds directly to eight promoters of the pimaricin cluster, as demonstrated by electrophoretic mobility shift assays. Assays with truncated forms of the protein revealed that the PAS domain does not mediate specificity or the distinct recognition of target genes, which rely on the DBD domain, but significantly reduces binding affinity up to 500-fold. Transcription start points were identified by 5-rapid amplification of cDNA ends, and the binding regions of PimM DBD were investigated by DNase I protection studies. In all cases, binding took place covering the ؊35 hexamer box of each promoter, suggesting an interaction of PimM and RNA polymerase to cause transcription activation. Information content analysis of the 16 sequences protected in target promoters was used to deduce the structure of the PimM-binding site. This site displays dyad symmetry, spans 14 nucleotides, and adjusts to the consensus TVGGGAWWTCCCBA. Experimental validation of this binding site was performed by using synthetic DNA duplexes. Binding of PimM to the promoter region of one of the polyketide synthase genes from the Streptomyces nodosus amphotericin cluster containing the consensus binding site was also observed, thus proving the applicability of the findings reported here to other antifungal polyketides.

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Section: Discussionmentioning

confidence: 99%

Molecular Control of Polyene Macrolide Biosynthesis

Santos‐Aberturas

Vicente

Guerra

et al. 2011

Journal of Biological Chemistry

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“…2,3 The two-component PhoRPhoP study was later extended to S. coelicolor, 4,5 Streptomyces natalensis, 6 Streptomyces clavuligerus 7 and some other Streptomyces species, including recently S. avermitilis. 8 The Streptomyces PhoR-PhoP system belongs to classIIIA of two-component systems.…”

Section: Genetic Basis Of Phosphate Control Of Metabolismmentioning

confidence: 99%

The master regulator PhoP coordinates phosphate and nitrogen metabolism, respiration, cell differentiation and antibiotic biosynthesis: comparison in Streptomyces coelicolor and Streptomyces avermitilis

2017

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Phosphate limitation is important for production of antibiotics and other secondary metabolites in Streptomyces. Phosphate control is mediated by the two-component system PhoR-PhoP. Following phosphate depletion, PhoP stimulates expression of genes involved in scavenging, transport and mobilization of phosphate, and represses the utilization of nitrogen sources. PhoP reduces expression of genes for aerobic respiration and activates nitrate respiration genes. PhoP activates genes for teichuronic acid formation and reduces expression of genes for phosphate-rich teichoic acid biosynthesis. In Streptomyces coelicolor, PhoP repressed several differentiation and pleiotropic regulatory genes, which affects development and indirectly antibiotic biosynthesis. A new bioinformatics analysis of the putative PhoP-binding sequences in Streptomyces avermitilis was made. Many sequences in S. avermitilis genome showed high weight values and were classified according to the available genetic information. These genes encode phosphate scavenging proteins, phosphate transporters and nitrogen metabolism genes. Among of the genes highlighted in the new studies was aveR, located in the avermectin gene cluster, encoding a LAL-type regulator, and afsS, which is regulated by PhoP and AfsR. The sequence logo for S. avermitilis PHO boxes is similar to that of S. coelicolor, with differences in the weight value for specific nucleotides in the sequence. INTRODUCTIONPhosphate is one of the essential nutrients of all living beings. In early studies, Martín and Demain 1 reported that, following phosphate starvation, there is a change in the metabolism of the antibiotic producing strains, which causes the slowdown of primary metabolism and triggers production of secondary metabolites. This early hypothesis has been largely confirmed by the research developed in the last few decades and is discussed in more detail below taking into account recent developments in the coordination of secondary metabolism. The aim of this article is to present an up-to-date integrative view of the PhoP regulation of metabolism in Streptomyces.

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“…Samples were disrupted with 2 cycles of 30 s at speed 6.5 in a Ribolyser instrument using the lysing matrix B (BIO 101) and RNA was extracted using an RNeasy mini kit (Qiagen) as described in Tunca et al 2007. RNA concentration and quality was checked using NanoDrop ND-1000 (Thermo Fisher Scientific).…”

Section: Reverse Transcriptase Pcr (Rt-pcr)mentioning

confidence: 99%

“…PhoP (Sola-Landa et al 2003;2005;Ghorbel et al 2006;Mendes et al 2007). The global phosphate response regulator PhoP and its target genes have been characterized in S.…”

Section: Introductionmentioning

confidence: 99%