Hybridization and DNA sequence analyses suggest an early evolutionary divergence of related biosynthetic gene sets encoding polyketide antibiotics and spore pigments in Streptomyces spp.

Blanco, Gloria; Brian, Paul; Pereda, Ana; Méndez, Cármen; Salas, JA; Kf, Chater

doi:10.1016/0378-1119(93)90352-4

Cited by 35 publications

(22 citation statements)

References 25 publications

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“…XSG-9 also hybridized with ORF4 of the gra PKS (CYC-DH). This clone was chosen for further characterization; the others may represent a PKS gene cluster for another polyketide, including perhaps a spore pigment PKS (8,12).…”

Section: Resultsmentioning

confidence: 99%

“…Comparisons of the amino acid sequences of the ORF1 and ORF2 products for the actinomycete aromatic polyketides (including spore pigments), using the PILEUP program of the Genetics Computer Group package, have suggested that a duplication of a progenitor of both ORFs was followed by divergence of the sequences within each ORF (8,24). A phylogenetic tree, involving a larger set of sequences including those of gris ORF1 and ORF2, is shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Sources of data are as follows: Gris, griseusin synthesis (the present paper); Ard, PKS with unknown function from the ardicin producer, Kibdelosporangium aridum (34); Mon, PKS with unknown function from the monensin producer, S. cinnamonensis (1); Tcm, tetracenomycin synthesis in S. glaucescens (4); Otc, oxytetracycline synthesis in S. rimosus (28); Gra, granaticin synthesis in S. violaceoruber (42); Hir, PKS of unknown function from Saccharopolyspora hirsuta (29); Act, actinorhodin synthesis in S. coelicolor (14); Fren, frenolicin synthesis in S. roseofulvus (7); WhiE, spore pigment synthesis in S. coelicolor (12); Sch, spore pigment synthesis in S. halstedii (8); and Cur, possible spore pigment synthesis in S. curacoi (3).…”

Section: Resultsmentioning

confidence: 99%

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Cloning, sequencing, and analysis of the griseusin polyketide synthase gene cluster from Streptomyces griseus

Bibb

Revill

et al. 1994

J Bacteriol

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A fragment of DNA was cloned from the Streptomyces griseus K-63 genome by using genes (act) for the actinorhodin polyketide synthase (PKS) of Streptomyces coelicolor as a probe. Sequencing of a 5.4-kb segment of the cloned DNA revealed a set of five gris open reading frames (ORFs), corresponding to the act PKS genes, in the following order: ORF1 for a ketosynthase, ORF2 for a chain length-determining factor, ORF3 for an acyl carrier protein, ORF5 for a ketoreductase, and ORF4 for a cyclase-dehydrase. Replacement of the gris genes with a marker gene in the S. griseus genome by using a single-stranded suicide vector propagated in Escherichia coli resulted in loss of the ability to produce griseusins A and B, showing that the five gris genes do indeed encode the type II griseusin PKS. These genes, encoding a PKS that is programmed differently from those for other aromatic PKSs so far available, will provide further valuable material for analysis of the programming mechanism by the construction and analysis of strains carrying hybrid PKSs.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Cloning, sequencing, and analysis of the griseusin polyketide synthase gene cluster from Streptomyces griseus

Bibb

Revill

et al. 1994

J Bacteriol

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“…Perhaps this resulted in a comparative excess of the whiE-ORFVIII substrate, mimicking the effect of blocking the reaction catalysed by whiE-ORFVIII. whiE-ORFVIII resembles S. halstedii scb-ORFC, and both gene products in turn resemble FADlinked hydroxylases (Blanco et al, 1993). Mutation of sch-ORFC changed the spore colour of S. balstedii from green to lilac (Blanco e t al., 1993).…”

Section: Roles Of the Whie Orfsmentioning

confidence: 99%

Ectopic expression of the Streptomyces coelicolor whiE genes for polyketide spore pigment synthesis and their interaction with the act genes for actinorhodin biosynthesis

Hopwood

1995

Microbiology

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The whiE gene cluster of Streptomyces coe/ico/or is normally expressed shortly before sporulation in the aerial mycelium, leading to production of the grey polyketide spore pigment. By placing the Mi€ genes under the control of the thiostrepton-inducible tipA promoter, they were artificially expressed on plasmids or in the chromosome during vegetative growth in a strain deleted for the act genes, which control biosynthesis of the polyketide antibiotic actinorhodin. Certain combinations of whiEORFI-VII led to production of mycelial pigments; these were exported into the medium when whi€-ORFI was absent, but poorly in its presence. Combined with comparative sequence data, the results allowed deductions to be made, or confirmed, about the normal roles of the eight known genes, whi€=ORFbVIII, as follows: whiFORFIII, IV, V encode the three components (ketosynthase, chain length factor and acyl carrier protein) of the whi€ 'minimal' polyketide synthase (PKS) needed for assembly of the carbon chain of the spore pigment precursor; whiE-ORFII, VI, VII are likely to be involved in cyclizations of the nascent carbon chain; whiE ORFVlll controls a late step in the spore pigment biosynthetic pathway, probably a hydroxylation; and whi€-ORFl may encode a protein needed for correct targeting or retention of spore pigment at an appropriate cellular location. In other experiments, genes encoding components of the act-PKS and whi€-PKS were artificially co-expressed. Each of the three whi€ minimal PKS subunit genes could complement lesions in the corresponding act-PKS genes to produce actinorhodin or related mycelial pigments, and each of the three act minimal PKS genes could complement lesions in the whi€ minimal PKS genes to cause spore pigmentation. Thus the two sets of PKS subunits, which are encoded by genes that have presumably diverged from a common ancestor, are still capable of biochemical 'crosstalk', but this is nonnally prevented because the gene sets are expressed in different 'tissues' of the differentiated Streptomyces colony. Ectopic expression of sets of whiE-PKS genes presumed to be sufficient to assemble a carbon chain caused inhibition of early growth of the strains, perhaps by causing interference with fatty acid biosynthesis; this yielded circumstantial evidence that the whi€-PKS gene products can also interact with those of the fatty acid synthase(s) of the organism. Abbreviations: ACP, acyl carrier protein; CLF, chain length factor; FAS, fatty acid synthase; KS, ketosynthase; PKS, polyketide synthase. 0001-9889 0 1995 SGM 2779Downloaded from www.microbiologyresearch.org by

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“…tested Blanco et al, 1993), bearing out the reasonable expectation that sporulation in all Streptomyces spp. would involve some common elements.…”

Section: Introductionmentioning

confidence: 68%

Functional and evolutionary implications of a survey of various actinomycetes for homologues of two Streptomyces coelicolor sporulation genes

Soliveri¹,

Vijgenboom²,

Granozzi³

et al. 1993

Journal of General Microbiology

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In Streptomyces coelicolor A3(2) the whiB and whiG genes are essential for sporulation, their deduced products being a possible transcriptional activator and an RNA polymerase sigma factor, respectively. In a survey of DNA from diverse actinomycetes by Southern blotting, all samples tested hybridized with whiB, but only those representing genera capable of producing sporulating aerial mycelium hybridized with whiG. It is postulated that whiB may play a more intimate role in hyphal fragmentation processes (including sporulation) than whiG. The whiB and whiG homologues (whiB-Stv and whiG-Stv) of Streptoverticillium griseocarneum were cloned and sequenced, and subjected to functional tests in S. coelicolor whiB and whiC mutants. The genes were closely similar, but not identical, to their S. coelicolor counterparts at the DNA and deduced protein levels, and both Stv. griseocarnum gene products could function well in S. coelicolor. However, studies with hybrid transcription units suggested that the promoter region of whiB-Stv is somewhat inefficient in S. coelicolor.

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Hybridization and DNA sequence analyses suggest an early evolutionary divergence of related biosynthetic gene sets encoding polyketide antibiotics and spore pigments in Streptomyces spp.

Cited by 35 publications

References 25 publications

Cloning, sequencing, and analysis of the griseusin polyketide synthase gene cluster from Streptomyces griseus

Cloning, sequencing, and analysis of the griseusin polyketide synthase gene cluster from Streptomyces griseus

Ectopic expression of the Streptomyces coelicolor whiE genes for polyketide spore pigment synthesis and their interaction with the act genes for actinorhodin biosynthesis

Functional and evolutionary implications of a survey of various actinomycetes for homologues of two Streptomyces coelicolor sporulation genes

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