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  • 1
    Electronic Resource
    Electronic Resource
    Organisation and functions of the actV A region of the actinorhodin biosynthetic gene cluster of Streptomyces coelicolor (1991)
    Springer
    Molecular genetics and genomics 230 (1991), S. 401-412 
    Details
    ISSN: 1617-4623
    Keywords: Antibiotic biosynthesis ; Antibiotic resistance ; Hydroxylases ; Streptomyces
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Sequence analysis of the actVA region of the actinorhodin biosynthetic gene cluster of Streptomyces coelicolor revealed a succession of six open reading frames (ORFs), all running in the same direction and extending over 5.32 kb. The protein product of actVA-ORF1 strongly resembles that of another gene, elsewhere in the act cluster (actII-ORF2), which codes for a trans-membrane protein previously implicated in actinorhodin export from the mycelium. This suggests that the two gene products may co-operate in actinorhodin export, perhaps being sufficient for self-protection of the organism against suicide. At least four of the other five ORFs are implicated in the control of the C-6 and C-8 ring-hydroxylation reactions, lacking in actVA mutants, that occur at middle to late stages in the actinorhodin biosynthetic pathway. This conclusion was reached by genetic mapping of actVA mutants to actVA-ORF3 and-ORF5 (and perhaps -ORF4), and by the finding of strong resemblances between the protein products of actVA-ORF2 and -ORF6 and the products of genes of the oxytetracycline or tetracenomycin gene clusters that have been implicated in ring-hydroxylation reactions in the biosynthesis of these other aromatic polyketide antibiotics.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00280297
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  • 2
    Electronic Resource
    Electronic Resource
    A mutation of Streptomyces lividans which prevents intraplasmid recombination has no effect on chromosomal recombination (1989)
    Springer
    Molecular genetics and genomics 220 (1989), S. 60-64 
    Details
    ISSN: 1617-4623
    Keywords: Plasmid ; Protoplast fusion ; Rec− mutations ; Recombination ; Streptomyces
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A mutation (rec-46) of Streptomyces lividans, previously shown to prevent (or greatly diminish) homologous and illegitimate intraplasmid recombination, was shown to have no effect on generalised chromosomal recombination occurring in matings or in protoplast fusions, nor to affect homologous recombination between a recombinant plasmid and the host chromosome. By comparison with Escherichia coli mutants defective in various aspects of recombination, the rec-46 mutation is similar to those in recF, recJ, recO and topA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00260856
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  • 3
    Electronic Resource
    Electronic Resource
    Transposition of IS117 (the Streptomyces coelicolor A3(2) mini-circle) to and from a cloned target site and into secondary chromosomal sites (1990)
    Springer
    Molecular genetics and genomics 224 (1990), S. 65-71 
    Details
    ISSN: 1617-4623
    Keywords: Streptomyces ; Transposable element ; ϕC31 phage ; Gene replacement
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary IS117, previously known as the 2.6 kb minicircle, is a transposable element found in Streptomyces coelicolor A3(2). It integrates predominantly into one preferred site when introduced into the closely related Streptomyces lividans 66, which lacks IS117. This preferred integration site was deleted from the S. lividans chromosome by replacement with an erythromycin resistance gene delivered by a ϕC31 phage vector. When IS117 was introduced into the resulting strain it integrated into many other sites, with some indication of site preference. By cloning a 200 by fragment centred on the preferred integration site onto a low copy number, self-transmissible Streptomyces plasmid derived from SCP2* it was shown that this sequence is sufficient to define the preferred site: IS117 integrates efficiently into this sequence from its preferred site in the host chromosome and at a lower frequency from the plasmid into the preferred site on the S. lividans chromosome.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00259452
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  • 4
    Electronic Resource
    Electronic Resource
    Transcriptional organization and regulation of an antibiotic export complex in the producing Streptomyces culture (1991)
    Springer
    Molecular genetics and genomics 228 (1991), S. 372-380 
    Details
    ISSN: 1617-4623
    Keywords: Streptomyces ; Antibiotic export ; Repressor ; S1 mapping ; Divergent promoters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Three open reading frames (ORFs) in the actII region of the actinorhodin biosynthetic gene cluster of Streptomyces coelicolor A3(2), which are involved in the export of the antibiotic are carried on two divergent transcripts. A monocistronic transcript carries actII-ORF1, encoding a putative repressor protein, and a bicistronic transcript codes for actII-ORF2 and -ORF3, whose products have been postulated to form an antibiotic export complex. The actll-ORF1 and actll-ORF2/3 transcripts each have a single promoter and the promoters for the two transcripts overlap. Both promoters are most active in cultures that have developed to the stage of actinorhodin production. The promoters resemble consensus promoters of the vegetative class in Escherichia coli and Streptomyces. We also demonstrate that these promoters are expressed in E. coli and use this finding to reveal a regulatory role for the repressor, using the xy/E reporter gene on promoter-probe shuttle vectors and regulated expression of the actII-ORF1 gene under control of Plac. The actII-ORF2/3 promoter is strongly repressed by the ORF1 product and the ORF1 product also represses its own promoter. The finding that the operator/promoter arrangement, and regulatory interconnection, of an antibiotic export/repressor gene pair in Streptomyces strikingly resemble those for tetracycline resistance in bacteria of clinical importance supports the hypothesis of an evolutionary origin of such genes in an ancestral actinomycete.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00260629
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  • 5
    Electronic Resource
    Electronic Resource
    A 2.6 kb DNA sequence of Streptomyces coelicolor A3(2) which functions as a transposable element (1986)
    Springer
    Molecular genetics and genomics 203 (1986), S. 79-88 
    Details
    ISSN: 1617-4623
    Keywords: Streptomyces ; Transposon ; Plasmid ; Integration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Streptomyces coelicolor A3(2) contains CCC DNA molecules, 2.6 kb in size, with an average copy number of less than one per ten chromosomes. Southern hybridisation revealed, in addition, two linear, integrated copies (A and B) of this “mini-circle” sequence per chromosome. The two integrated copies have similar (if not identical) ends and are present in the same locations in various S. coelicolor A3(2) derivatives. The mini-circle sequence is absent from S. lividans 66 and S. violaceolatus ISP5438 and from several Streptomyces species less closely related to S. coelicolor A3(2). None of a variety of Streptomyces plasmids tested contained homology to the mini-circle sequence. When a 1.8 kb fragment of the mini-circle lacking the ends of the integrated copies was inserted into KC515 (a derivative of the temperate phage ϕC31 which is unable to lysogenise host strains by the natural route because the phage attachment site has been deleted) the resulting phage lysogenised S. coelicolor A3(2) (integrating into the genome of this host by homologous recombination with resident minicircle sequences) but not S. lividans or a variety of other ϕC31 hosts. In contrast, a KC515 derivative (KC591) carrying the entire 2.6 kb mini-circle sequence linearised at its single BclI site (and therefore containing the integration site of the free mini-circle) lysogenised not only S. coelicolor A3(2) but also S. lividans 66 and most other strains normally lysogenised by ϕC31. The KC591 lysogens of the eight Streptomyces species tested contained a linear, integrated prophage with termini apparently identical to those of the linear mini-cricle copies of S. coelicolor. In S. lividans, KC591 integrated preferentially at a site apparently homologous to the site occupied by mini-circle sequence A in S. coelicolor A3(2) strains, but integration into secondary sites also occurred.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00330387
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  • 6
    Electronic Resource
    Electronic Resource
    Physical and genetic characterisation of the gene cluster for the antibiotic actinorhodin inStreptomyces coelicolor A3(2) (1986)
    Springer
    Molecular genetics and genomics 205 (1986), S. 66-73 
    Details
    ISSN: 1617-4623
    Keywords: Streptomyces ; Polyketide antibiotic ; Mutational cloning ; ϕC31 phage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We determined the physical and transcriptional organisation of the set of previously cloned biosynthetic genes involved in the production of the polyketide antibiotic actinorhodin byStreptomyces coelicolor A3(2). Complementation and mutational cloning analyses (in part using new ϕC31 phage vectors incorporating a transcriptional terminator to block transcription from vector promoters into the cloned DNA) indicate that all the biosynthetic genes, including at least one regulatory (activator) gene, are clustered in a chromosomal region of about 26 kb. The genes are organised in at least four separate transcription units, ranging in size from 1 kb for the class III gene, to a polycistronic transcript of at least 5 kb for the class I, VII and IV genes. Indirect evidence shows that resistance to actinorhodin is also determined by the cloned DNA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02428033
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  • 7
    Electronic Resource
    Electronic Resource
    Mutation and cloning of clustered Streptomyces genes essential for sulphate metabolism (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 415-423 
    Details
    ISSN: 1617-4623
    Keywords: Streptomyces ; Selenate resistance ; Cysteine auxotrophy ; Sulphate permease ; ATP sulphurylase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A range of mutants auxotrophic for cysteine (cys) and resistant to selenate (sel) were isolated from many Streptomyces strains but chiefly from S. coelicolor A3(2) and S. lividans 66. Two of the classes of sel/cys mutants probably contained simple biochemical lesions of sulphate permease (selC) and ATP sulphurylase (selA) activities, while a further two classes (selD and selE) were pleiotropic and possibly regulatory. Most classes of sel mutations were clustered around the cysD locus of S. coelicolor. Segments of chromosomal DNA cloned from S. coelicolor, S. cattleya and S. clavuligerus and able to complement various sel/cys mutations allowed the relative positions of these mutations and the cysC and cysD mutations of S. coelicolor to be determined. The sel/cys DNA can be used for two-way selection: Cys+Sels↔Cys-Selr.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00425694
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  • 8
    Electronic Resource
    Electronic Resource
    afsB stimulates transcription of the actinorhodin biosynthetic pathway in Streptomyces coelicolor A3(2) and Streptomyces lividans (1989)
    Springer
    Molecular genetics and genomics 215 (1989), S. 355-357 
    Details
    ISSN: 1617-4623
    Keywords: Streptomyces ; Secondary metabolism ; Transcriptional stimulation ; afsB ; Pleiotropic regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The pleiotropic regulatory gene, afsB, from Streptomyces coelicolor A3(2), possibly encoding a DNA-binding protein, is required for actinorhodin production in this organism. Northern blot hybridization using a DNA fragment covering part of the set of cloned actinorhodin biosynthetic gene cluster (act) as the probe showed lack of the act transcripts in an afsB-negative mutant of S. coelicolor A3(2); the transcripts were restored on introduction of a cloned afsB gene. Introduction of the cloned afsB gene into Streptomyces lividans stimulated transcription of the act genes under conditions in which they are normally silent in this strain, leading to production of actinorhodin in large quantity. These data show that afsB exerts its positive regulatory effect by means of transcriptional stimulation of its target genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00339742
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