• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.480-484
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• 2010

AfsR2 is a global regulatory protein that stimulates antibiotic biosynthesis in both Streptomyces lividans and S. coelicolor. Previously, various afsR2-dependent genes including a putative abaA-like regulatory gene, SCO4677, were identified through comparative DNA microarray analysis. To further identify the putative SCO4677-dependent proteins, the comparative proteomics-driven approach was applied to the SCO4677-overexpressing strains of S. lividans and S. coelicolor along with the wild-type strains. The 2D gel electrophoresis gave approximately 277 protein spots for S. lividans and 207 protein spots for S. coelicolor, showing different protein expression patterns between the SCO4677-overexpressing strains and the wild-type strains. Further MALDI-TOF analysis revealed that only 18 proteins exhibited similar expression patterns in both S. lividans and S. coelicolor, suggesting that the SCO4677 could encode an abaA-like regulator that controls a few cross-species common proteins as well as many species-specific proteins in Streptomyces species.

• KSBB Journal
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• v.19 no.6 s.89
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• pp.462-466
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• 2004

Mycothiol (MSH), a low molecular antioxidant thiol compound, was purified and analyzed from Streptomyns coelicolor A[3]2 by the monobromobimane fluorescence detection method modified by this lab. Through HPLC chromatpgram, MSH fraction was obtained following the elution time of standard MSH (donated by Dr. Robert C. Fahey). That MSH showed the highest concentration among the thiol compounds contained in the cell indicated that MSH was the key thiol compound having antioxidant activity. To understand the role of gene of inositol monophosphatase (I-1-Pase) involved in the MSH biosynthesis, it was isolated from S. coelicolor A(3)2 and cloned and overexpressed in the Escherichia coli. The expressed I-1-Pase was purified through Ni-NTA column. The soluble protein consisted of 281 amino acids, and the molecular weight was 32 kDa. I-1-Pase of S. coelicolor A(3)2 had the sequence homology with those of human and E. coli by 24 and $25\%$, respectively, and had two conserved domains (mofif A and motif B) which were typical of I-1-Pase.

• Korean Journal of Microbiology
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• v.32 no.4
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• pp.264-270
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• 1994

A gene coding for a novel putative $\sigma$ factor of RNA polymerase has been identified from Streptomyces coelicolor A3(2) using Escherichia coli rpoS gene fragment as a probe. The 486 bp rpoS gene fragment was amplified from E. coli genomic DNA by PCR with two synthetic oligonucleotides, the sequences of which were deduced from the amino acid sequences in the regions 2.3 and 4.2 conserved among various bacterial factors. When E. coli genomic DNA fragments were hybridized with cloned rpoS probe, only one band corresponding to rpoS gene (3.2 kb PvuII fragment or 2.3 kb KpnI fragment) was detected. In S. coelicolor, however, two bands were detected both in PvuII digested DNA and SalI digested DNA. 3.5 kb PvuII fragment which binds the rpoS gene probe was cloned (pMS1) from the sublibrary, and the nucleotide sequences of 1.0 kb BamH'/HincII subclone (pBH2) was partially determined. The nucleotide sequences revealed extensive similarity to other $\sigma$ factor genes of S. coelicolor (hrdA, hrdB, hrdC, hrdD), S. aureofaciens (hrdA, hrdB, hrdC, hrdD), Synechococcus species, Pseudomonas aeruginosa, Stigmatella aurantiaca, and Anabaena species. The nucleotide sequences in regions 1.2 and 4 were compared with the corresponding regions of 5 known ${\sigma}$ factor genes of S. coelicolor by multiple alignment. It turned out that the cloned gene is most closely related to hrdA showing 88% amino acid similarity in region 1.2 and 75% in region 4.

• Genomics & Informatics
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• v.6 no.1
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• pp.44-49
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• 2008

Protein kinase C (PKC) is a family of kinases involved in the transduction of cellular signals that promote lipid hydrolysis. PKC plays a pivotal role in mediating cellular responses to extracellular stimuli involved in proliferation, differentiation and apoptosis. Comparative analysis of the PKC-${\alpha},{\beta},{\varepsilon}$ isozymes of 200 recently sequenced microbial genomes was carried out using variety of bioinformatics tools. Diversity and evolution of PKC was determined by sequence alignment. The ser/thr protein kinases of Streptomyces coelicolor A3 (2), is the only bacteria to show sequence alignment score greater than 30% with all the three PKC isotypes in the sequence alignment. S.coelicolor is the subject of our interest because it is notable for the production of pharmaceutically useful compounds including anti-tumor agents, immunosupressants and over two-thirds of all natural antibiotics currently available. The comparative analysis of three human isotypes of PKC and Serine/threonine protein kinase of S.coelicolor was carried out and possible mechanism of action of PKC was derived. Our analysis indicates that Serine/ threonine protein kinase from S. coelicolor can be a good candidate for potent anti-tumor agent. The presence of three representative isotypes of the PKC super family in this organism helps us to understand the mechanism of PKC from evolutionary perspective.

• Microbiology and Biotechnology Letters
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• v.23 no.6
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• pp.678-686
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• 1995

SecY is a central component of the protein export machinery that mediate the translocation of secretory proteins across the plasma membrane of Escherichia coli. In order to study the mechanism of protein secretion in Streptomyces, we have done cloning and sequencing of the Streptomyces coelicolor secY gene by using polymerase chain reaction method. The nucleotide sequence of the gene for SecY from S. coelicolor showed over 58% identity to that of M. luteus. The deduced amino acid sequences were highly homologous to those of other known SecY polypeptides, all having the potential to form 10 transmembrane segments, and especially second, fifth, and tenth segments were particularly conserved, sharing greater than 75% identity with W. lute s SecY. We propose that the conserved membrane-spanning segments actively participate in protein export. In B. subtilis and E. coli, the secY gene is a part of the spc operon, is preceded by the gene coding for ribosomal protein L15, and is likety coupled transcriptionally and translationally to the upstream L15 gene. In the other hand, secY gene of S. coelicolor and M. luteus have its own promoter region, are coupled translationally with adk gene and pr sented in adk operon.

• Journal of Microbiology
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• v.42 no.1
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• pp.64-67
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• 2004

The pellets from a culture of Streptomyces coelicolor A3(2) that were submerged shaken were disintegrated into numerous hyphal fragments by DNase treatment. The pellets were increasingly dispersed by hyaluronidase treatment, and mycelial fragments were easily detached from the pellets. The submerged mycelium grew by forming complexes with calcium phosphate precipitates or kaolin, a soil particle. Therefore, the pellet formation of Streptomyces coelicolor A3(2) can be considered a biofilm formation, including the participation of adhesive extracellular polymers and the insoluble substrates.

• Journal of Microbiology and Biotechnology
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• v.22 no.6
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• pp.736-741
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• 2012

In this study, we analyzed the oxidative stress response of wblA ($\underline{w}$hi$\underline{B}$-$\underline{l}$ike gene $\underline{A}$, SCO3579), which was previously shown to be a global antibiotic down-regulator in Streptomyces coelicolor. Ever since a WblA ortholog named WhcA in Corynebacterium glutamicum was found to play a negative role in the oxidative stress response, S. coelicolor wblA has been proposed to have a similar effect. A wblA-deletion mutant exhibited a less sensitive response to oxidative stress induced by diamide present in solid plate culture. Using real-time RT-PCR analysis, we also compared the transcription levels of oxidative stress-related genes, including sodF, sodF2, sodN, trxB, and trxB2, between S. coelicolor wild type and a wblA-deletion mutant in the presence or absence of oxidative stress. Target genes were expressed higher in the wblA-deletion mutant compared with wild type, both in the absence and presence of oxidative stress. Moreover, expression of these target genes in S. coelicolor wild type was stimulated only in the presence of oxidative stress, suggesting that WblA plays a negative role in the oxidative stress response of S. coelicolor, similar to that of C. glutamicum WhcA, through the transcriptional regulation of oxidative stress-related genes.

• Journal of Microbiology and Biotechnology
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• v.31 no.11
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• pp.1591-1600
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• 2021

Streptomyces coelicolor is a filamentous soil bacterium producing several kinds of antibiotics. S. coelicolor abs8752 is an abs (antibiotic synthesis deficient)-type mutation at the absR locus; it is characterized by an incapacity to produce any of the four antibiotics synthesized by its parental strain J1501. A chromosomal DNA fragment from S. coelicolor J1501, capable of complementing the abs- phenotype of the abs8752 mutant, was cloned and analyzed. DNA sequencing revealed that two complete ORFs (SCO6992 and SCO6993) were present in opposite directions in the clone. Introduction of SCO6992 in the mutant strain resulted in a remarkable increase in the production of two pigmented antibiotics, actinorhodin and undecylprodigiosin, in S. coelicolor J1501 and abs8752. However, introduction of SCO6993 did not show any significant difference compared to the control, suggesting that SCO6992 is primarily involved in stimulating the biosynthesis of antibiotics in S. coelicolor. In silico analysis of SCO6992 (359 aa, 39.5 kDa) revealed that sequences homologous to SCO6992 were all annotated as hypothetical proteins. Although a metalloprotease domain with a conserved metal-binding motif was found in SCO6992, the recombinant rSCO6992 did not show any protease activity. Instead, it showed very strong β-glucuronidase activity in an API ZYM assay and toward two artificial substrates, p-nitrophenyl-β-D-glucuronide and AS-BI-β-D-glucuronide. The binding between rSCO6992 and Zn²⁺ was confirmed by circular dichroism spectroscopy. We report for the first time that SCO6992 is a novel protein with β-glucuronidase activity, that has a distinct primary structure and physiological role from those of previously reported β-glucuronidases.

• Journal of Microbiology and Biotechnology
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• v.16 no.5
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• pp.799-803
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• 2006

It was reported that an accumulation of Sadenosyl-L-methionine increases production of actinorhodin in Streptomyces lividans and induces antibiotic biosynthetic genes. We also obtained the same result in Streptomyces coelicolor A3(2). Therefore, in order to identify proteins changed by the addition of S-adenosyl-L-methionine in S. coelicolor A3(2), LC/MS/MS analyses were carried out. Thirteen proteins that were not observed in the control were found.

• Korean Journal of Microbiology
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• v.43 no.1
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• pp.72-75
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• 2007

Using co-immunoprecipitation, we identified proteins interacting with Streptomyces coelicolor RNase ES, an ortholog of Escherichia coli RNase E that plays a major role in RNA decay and processing. Polyphosphate kinase and a homolog of exoribonuclease polynucleotide phosphorylase, guanosine pentaphosphate synthetase I that use inorganic phophate were co-precipitated with RNase E, indicating a possibility of S. coelicolor RNase ES to form a multiprotein complex called degradosome, which has been shown to be formed by RNase E in E. coli. Polynucleotide phophorylase proteins from these two phylogenetically distantly related bacteria species showed similar RNA cleavage action in vitro. These results imply the ability of RNase ES to form a multiprotein complex that has structurally and functionally similar to that of E. coli degradosome.