Cloning of the conserved regulatory operon by its aerial mycelium-inducing activity in an amfR mutant of Streptomyces griseus

Komatsu, Mamoru; Kuwahara, Yoshitaka; Hiroishi, Ayako; Hosono, Kuniaki; Beppu, Teruhiko; Uéda, Kenji

doi:10.1016/s0378-1119(03)00405-0

Cited by 25 publications

(38 citation statements)

References 27 publications

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“…Protected fragments were analyzed on a 6% polyacrylamide gel. The quality of RNA used for low-resolution analysis was checked by a control assay for hrdB, encoding a major sigma factor, using a probe described previously (19).…”

Section: Methodsmentioning

confidence: 99%

Light-Induced Carotenogenesis inStreptomyces coelicolorA3(2): Identification of an Extracytoplasmic Function Sigma Factor That Directs Photodependent Transcription of the Carotenoid Biosynthesis Gene Cluster

et al. 2005

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Carotenoids are produced by a variety of organisms, but the mechanisms that regulate gene expression leading to carotenoid biosynthesis have been characterized for only a few organisms. In this study, we found that Streptomyces coelicolor A3(2), a gram-positive filamentous bacterium, produces carotenoids under blue light induction. The carotenoid fraction isolated from the cell extract contained multiple compounds, including isorenieratene and ␤-carotene. The carotenoid biosynthesis gene cluster of S. coelicolor consists of two convergent operons, crtEIBV and crtYTU, as previously shown for Streptomyces griseus. The crtEIBV null mutant completely lost its ability to produce carotenoids. The crt gene cluster is flanked by a regulatory region that consists of two divergent operons, litRQ and litSAB. The lit (light-induced transcription) genes encode a MerR-type transcriptional regulator (LitR), a possible oxidoreductase (LitQ), an extracytoplasmic function sigma factor ( LitS ), a putative lipoprotein (LitA), and a putative anti-sigma factor (LitB). S1 protection assay revealed that the promoters preceding crtE (PcrtE), crtY (PcrtY), litR (PlitR), and litS (PlitS) are activated upon illumination. A litS mutant lost both the ability to produce carotenoids and the activities of PcrtE, PcrtY, and PlitS, which suggested that LitS directs light-induced transcription from these promoters. An RNA polymerase holocomplex containing purified LitS recombinant protein generated specific PcrtE and PcrtY transcripts in an in vitro runoff transcriptional assay. A litR mutant that had an insertion of the kanamycin resistance gene was defective both in the ability to produce carotenoids and in all of the light-dependent promoter activities. Overexpression of litS resulted in constitutive carotenoid production in both the wild type and the litR mutant. These results indicate that LitS acts as a light-induced sigma factor that directs transcription of the crt biosynthesis gene cluster, whose activity is controlled by an unknown LitR function. This is the first report to describe light-inducible gene expression in Streptomyces.

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Section: Methodsmentioning

confidence: 99%

Light-Induced Carotenogenesis inStreptomyces coelicolorA3(2): Identification of an Extracytoplasmic Function Sigma Factor That Directs Photodependent Transcription of the Carotenoid Biosynthesis Gene Cluster

et al. 2005

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“…Bentley et al (26) noted the presence in S. coelicolor of 13 paralogous sets of four genes, which they called conservons, that encode membrane-bound complexes resembling eukaryotic G protein-coupled regulatory systems (234). In each set, the first gene, cvnA, encodes a sensor kinase-like ATPase, and the fourth, cvnD, encodes a guanine nucleotide-binding GTPase.…”

Section: Comparative Genomics Of Mycelial Actinobacterial Genomesmentioning

confidence: 99%

Genomics ofActinobacteria: Tracing the Evolutionary History of an Ancient Phylum

Ventura

Canchaya

Tauch

et al. 2007

Microbiol Mol Biol Rev

933

638

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SUMMARY Actinobacteria constitute one of the largest phyla among Bacteria and represent gram-positive bacteria with a high G+C content in their DNA. This bacterial group includes microorganisms exhibiting a wide spectrum of morphologies, from coccoid to fragmenting hyphal forms, as well as possessing highly variable physiological and metabolic properties. Furthermore, Actinobacteria members have adopted different lifestyles, and can be pathogens (e.g., Corynebacterium, Mycobacterium, Nocardia, Tropheryma, and Propionibacterium), soil inhabitants (Streptomyces), plant commensals (Leifsonia), or gastrointestinal commensals (Bifidobacterium). The divergence of Actinobacteria from other bacteria is ancient, making it impossible to identify the phylogenetically closest bacterial group to Actinobacteria. Genome sequence analysis has revolutionized every aspect of bacterial biology by enhancing the understanding of the genetics, physiology, and evolutionary development of bacteria. Various actinobacterial genomes have been sequenced, revealing a wide genomic heterogeneity probably as a reflection of their biodiversity. This review provides an account of the recent explosion of actinobacterial genomics data and an attempt to place this in a biological and evolutionary context.

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“…4A). The four genes immediately upstream of, and overlapping, SCO6939 (cvnA8 to cvnD8) have been designated a conservon to reflect the conserved clustering and probable operon structure of these genes and the 12 other paralogous clusters in the genome (4), and the cvn9 cluster has recently been implicated in aerial mycelium development in both S. coelicolor and Streptomyces griseus (22). Analysis of the sequence of the SCO6939 product by a BLAST search (2) indicated that its closest homologs were in Streptomyces, including SCO6798 (SC1A2.07) in S. coelicolor (whose corresponding gene is also located immediately downstream of a conservon, cvnA7 to cvnD7).…”

Section: Mutagenesismentioning

confidence: 99%

Novel Genes That Influence Development inStreptomyces coelicolor

et al. 2004

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Filamentous soil bacteria of the genus Streptomyces carry out complex developmental cycles that result in sporulation and production of numerous secondary metabolites with pharmaceutically important activities. To further characterize the molecular basis of these developmental events, we screened for mutants of Streptomyces coelicolor that exhibit aberrant morphological differentiation and/or secondary metabolite production. On the basis of this screening analysis and the subsequent complementation analysis of the mutants obtained we assigned developmental roles to a gene involved in methionine biosynthesis (metH) and two previously uncharacterized genes (SCO6938 and SCO2525) and we reidentified two previously described developmental genes (bldA and bldM). In contrast to most previously studied genes involved in development, the genes newly identified in the present study all appear to encode biosynthetic enzymes instead of regulatory proteins. The MetH methionine synthase appears to be required for conversion of aerial hyphae into chains of spores, SCO6938 is a probable acyl coenzyme A dehydrogenase that contributes to the proper timing of aerial mycelium formation and antibiotic production, and SCO2525 is a putative methyltransferase that influences various aspects of colony growth and development.

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Cloning of the conserved regulatory operon by its aerial mycelium-inducing activity in an amfR mutant of Streptomyces griseus

Cited by 25 publications

References 27 publications

Light-Induced Carotenogenesis inStreptomyces coelicolorA3(2): Identification of an Extracytoplasmic Function Sigma Factor That Directs Photodependent Transcription of the Carotenoid Biosynthesis Gene Cluster

Light-Induced Carotenogenesis inStreptomyces coelicolorA3(2): Identification of an Extracytoplasmic Function Sigma Factor That Directs Photodependent Transcription of the Carotenoid Biosynthesis Gene Cluster

Genomics ofActinobacteria: Tracing the Evolutionary History of an Ancient Phylum

Novel Genes That Influence Development inStreptomyces coelicolor

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