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Pleiotropic role of the Sco1/SenC family copper chaperone in the physiology of Streptomyces.

Fujimoto M, Yamada A, Kurosawa J, Kawata A, Beppu T, Takano H, Ueda K - Microb Biotechnol (2011)

Bottom Line: The scoC mutant of S. griseus was also defective in the extracellular activity oxidizing N,N'-dimethyl-p-phenylenediamine sulfate.Addition of 10 µM CuSO(4) repressed the activity of the conserved promoter preceding scoA and caused phenylalanine auxotrophy in some Streptomyces spp. probably because of the repression of pheA; pheA encodes prephenate dehydratase, which is located at the 3' terminus of the putative operon structure.Overall, the evidence indicates that Sco is crucial for the utilization of copper under a low-copper condition and for the activation of the multiple Cu(2+) -containing oxidases that play divergent roles in the complex physiology of Streptomyces.

View Article: PubMed Central - PubMed

Affiliation: Life Science Research Center, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Japan.

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Phenotype of the scoC mutant of S. coelicolor A3(2). A. Macroscopic observation of the wild‐type strain (WT) and the scoC mutant (ΔscoC). Colonies were photographed at days 2 and 5 from the top (T) and bottom (B) to show the occurrence of aerial mycelium and the pigment antibiotic actinorhodin respectively. Strains were grown at 28°C on Bennett's solid medium containing 1% glucose (Glc) and maltose (Mal) without and with 10 µM CuSO4. Colonies forming aerial mycelia appear white, whereas those forming only substrate mycelium appear yellow or light brown. The purple or blue colour is due to the production of the pigmented antibiotic, actinorhodin. Streptomyces coelicolor also produces an intracellular red pigment undecylprodigiosin. B. Scanning electron microscopy observation of the wild type and the scoC mutant grown for 3 days on Bennett's/glucose solid medium. Bar, 2 µm. C. Macroscopic observation of the genetically complemented scoC mutant. The scoC mutants harbouring the empty integration vector (pKU460), and pKU460 containing the intact (pKU460‐scoC) and mutated [pKU460‐scoC (C86S/C90S)]scoC‐coding sequence, were grown on Bennett's/maltose medium for 2 days. The wild type harbouring pKU460 is also shown.
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f2: Phenotype of the scoC mutant of S. coelicolor A3(2). A. Macroscopic observation of the wild‐type strain (WT) and the scoC mutant (ΔscoC). Colonies were photographed at days 2 and 5 from the top (T) and bottom (B) to show the occurrence of aerial mycelium and the pigment antibiotic actinorhodin respectively. Strains were grown at 28°C on Bennett's solid medium containing 1% glucose (Glc) and maltose (Mal) without and with 10 µM CuSO4. Colonies forming aerial mycelia appear white, whereas those forming only substrate mycelium appear yellow or light brown. The purple or blue colour is due to the production of the pigmented antibiotic, actinorhodin. Streptomyces coelicolor also produces an intracellular red pigment undecylprodigiosin. B. Scanning electron microscopy observation of the wild type and the scoC mutant grown for 3 days on Bennett's/glucose solid medium. Bar, 2 µm. C. Macroscopic observation of the genetically complemented scoC mutant. The scoC mutants harbouring the empty integration vector (pKU460), and pKU460 containing the intact (pKU460‐scoC) and mutated [pKU460‐scoC (C86S/C90S)]scoC‐coding sequence, were grown on Bennett's/maltose medium for 2 days. The wild type harbouring pKU460 is also shown.

Mentions: To study the role of the Sco1/SenC family copper chaperone protein, a marker‐less knockout mutant for the corresponding coding sequence (scoC) was generated with respect to S. coelicolor A3(2) (Fig. 2). As shown in Fig. 2A (upper panels), aerial mycelium formation and pigment antibiotic production in the scoC mutant of S. coelicolor was delayed significantly. The wild type formed aerial mycelia and produced pigment antibiotics on day 2 on Bennett's medium supplied with 1% glucose, whereas the mutant was pale brown in colour and formed only vegetative hyphae. Scanning electron microscope observation (Fig. 2B) showed the presence of abundant aerial mycelia and spores in the wild type but only substrate hyphae in the scoC mutant. However, on day 5, the mutant formed aerial mycelia and pigment to the same extent as did the wild type (Fig. 2A). The delay in development of the scoC mutant was recovered by supplying 10 µM CuSO4 to the culture medium (Fig. 2A, lower panels) or by introducing an intact scoC using an integration vector (Fig. 2C). The delay of development was also observed when the mutant was cultured on the medium supplied with maltose (Fig. 2A), indicating that the phenotype is not specific to glucose. Similar delay of development was observed with respect to the scoC mutant of S. griseus (data not shown). These results indicate that the copper utilization by ScoC and subsequent activation of some copper‐dependent function(s) is crucial for the initiation of developmental growth.


Pleiotropic role of the Sco1/SenC family copper chaperone in the physiology of Streptomyces.

Fujimoto M, Yamada A, Kurosawa J, Kawata A, Beppu T, Takano H, Ueda K - Microb Biotechnol (2011)

Phenotype of the scoC mutant of S. coelicolor A3(2). A. Macroscopic observation of the wild‐type strain (WT) and the scoC mutant (ΔscoC). Colonies were photographed at days 2 and 5 from the top (T) and bottom (B) to show the occurrence of aerial mycelium and the pigment antibiotic actinorhodin respectively. Strains were grown at 28°C on Bennett's solid medium containing 1% glucose (Glc) and maltose (Mal) without and with 10 µM CuSO4. Colonies forming aerial mycelia appear white, whereas those forming only substrate mycelium appear yellow or light brown. The purple or blue colour is due to the production of the pigmented antibiotic, actinorhodin. Streptomyces coelicolor also produces an intracellular red pigment undecylprodigiosin. B. Scanning electron microscopy observation of the wild type and the scoC mutant grown for 3 days on Bennett's/glucose solid medium. Bar, 2 µm. C. Macroscopic observation of the genetically complemented scoC mutant. The scoC mutants harbouring the empty integration vector (pKU460), and pKU460 containing the intact (pKU460‐scoC) and mutated [pKU460‐scoC (C86S/C90S)]scoC‐coding sequence, were grown on Bennett's/maltose medium for 2 days. The wild type harbouring pKU460 is also shown.
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f2: Phenotype of the scoC mutant of S. coelicolor A3(2). A. Macroscopic observation of the wild‐type strain (WT) and the scoC mutant (ΔscoC). Colonies were photographed at days 2 and 5 from the top (T) and bottom (B) to show the occurrence of aerial mycelium and the pigment antibiotic actinorhodin respectively. Strains were grown at 28°C on Bennett's solid medium containing 1% glucose (Glc) and maltose (Mal) without and with 10 µM CuSO4. Colonies forming aerial mycelia appear white, whereas those forming only substrate mycelium appear yellow or light brown. The purple or blue colour is due to the production of the pigmented antibiotic, actinorhodin. Streptomyces coelicolor also produces an intracellular red pigment undecylprodigiosin. B. Scanning electron microscopy observation of the wild type and the scoC mutant grown for 3 days on Bennett's/glucose solid medium. Bar, 2 µm. C. Macroscopic observation of the genetically complemented scoC mutant. The scoC mutants harbouring the empty integration vector (pKU460), and pKU460 containing the intact (pKU460‐scoC) and mutated [pKU460‐scoC (C86S/C90S)]scoC‐coding sequence, were grown on Bennett's/maltose medium for 2 days. The wild type harbouring pKU460 is also shown.
Mentions: To study the role of the Sco1/SenC family copper chaperone protein, a marker‐less knockout mutant for the corresponding coding sequence (scoC) was generated with respect to S. coelicolor A3(2) (Fig. 2). As shown in Fig. 2A (upper panels), aerial mycelium formation and pigment antibiotic production in the scoC mutant of S. coelicolor was delayed significantly. The wild type formed aerial mycelia and produced pigment antibiotics on day 2 on Bennett's medium supplied with 1% glucose, whereas the mutant was pale brown in colour and formed only vegetative hyphae. Scanning electron microscope observation (Fig. 2B) showed the presence of abundant aerial mycelia and spores in the wild type but only substrate hyphae in the scoC mutant. However, on day 5, the mutant formed aerial mycelia and pigment to the same extent as did the wild type (Fig. 2A). The delay in development of the scoC mutant was recovered by supplying 10 µM CuSO4 to the culture medium (Fig. 2A, lower panels) or by introducing an intact scoC using an integration vector (Fig. 2C). The delay of development was also observed when the mutant was cultured on the medium supplied with maltose (Fig. 2A), indicating that the phenotype is not specific to glucose. Similar delay of development was observed with respect to the scoC mutant of S. griseus (data not shown). These results indicate that the copper utilization by ScoC and subsequent activation of some copper‐dependent function(s) is crucial for the initiation of developmental growth.

Bottom Line: The scoC mutant of S. griseus was also defective in the extracellular activity oxidizing N,N'-dimethyl-p-phenylenediamine sulfate.Addition of 10 µM CuSO(4) repressed the activity of the conserved promoter preceding scoA and caused phenylalanine auxotrophy in some Streptomyces spp. probably because of the repression of pheA; pheA encodes prephenate dehydratase, which is located at the 3' terminus of the putative operon structure.Overall, the evidence indicates that Sco is crucial for the utilization of copper under a low-copper condition and for the activation of the multiple Cu(2+) -containing oxidases that play divergent roles in the complex physiology of Streptomyces.

View Article: PubMed Central - PubMed

Affiliation: Life Science Research Center, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Japan.

Show MeSH
Related in: MedlinePlus