Physical identification of a chromosomal locus encoding biosynthetic genes for the lipopeptide calcium-dependent antibiotic (CDA) of Streptomyces coelicolor A3(2)

Chong, Pei Pei; Podmore, Sylvia M.; Kieser, Helen M.; Redenbach, Matthias; Turgay, Kürşad; Marahiel, Mohamed A.; Hopwood, David A.; Smith, Colin P.

doi:10.1099/00221287-144-1-193

Cited by 59 publications

(57 citation statements)

References 28 publications

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“…By combining microarray data with information obtained by mapping, annotation, and sequencing of the S. coelicolor chromosome, we inferred the extent of known antibiotic biosynthetic clusters and also discovered new groupings of physically contiguous and coordinately expressed genes that may have related functions. Additionally, our mutational analysis identified chromosomally dispersed sets of ORFs whose expression was Contiguous genes whose expression is coordinately regulated throughout growth as determined by GABRIEL proband-based and continuity/gap rules are indicated as red and cda clusters, and these are displayed on linear maps (C, red) and (D, cda) constructed from sequence data available from the Sanger Centre and previous studies (Malpartida et al 1990;White and Bibb 1997;Chong et al 1998;Anderson et al 2001). Arrows indicate the extent and direction of putative ORFs.…”

Section: Discussionmentioning

confidence: 99%

“…An ∼35-kb segment of this region has been shown be essential for biosynthesis of CDA (Chong et al 1998); however, the full extent of the CDA locus has not been defined previously. Our analysis of transcription of genes within the CDA contig, which includes four overlapping cosmids sequenced by the Sanger Centre, has identified a continuous stretch of the S. coelicolor chromosome whose expression is temporally coordinated with known CDA genes (Fig.…”

Section: Microarray Analysis Of Streptomyces Transcription Genes and Dementioning

confidence: 99%

“…Genetic studies have mapped the locus encoding the lipopeptide CDA to a region later deduced to correspond to an ∼88 kb region of the S. coelicolor chromosome (Hopwood and Wright 1983;Chong et al 1998). An ∼35-kb segment of this region has been shown be essential for biosynthesis of CDA (Chong et al 1998); however, the full extent of the CDA locus has not been defined previously.…”

Section: Microarray Analysis Of Streptomyces Transcription Genes and Dementioning

confidence: 99%

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Global analysis of growth phase responsive gene expression and regulation of antibiotic biosynthetic pathways inStreptomyces coelicolorusing DNA microarrays

Huang¹,

Lih²,

Pan³

et al. 2001

Genes Dev.

195

188

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The eubacterial species Streptomyces coelicolor proceeds through a complex growth cycle in which morphological differentiation/development is associated with a transition from primary to secondary metabolism and the production of antibiotics. We used DNA microarrays and mutational analysis to investigate the expression of individual genes and multigene antibiotic biosynthetic pathways during these events. We identified expression patterns in biosynthetic, regulatory, and ribosomal protein genes that were associated highly specifically with particular stages of development. A knowledge-based algorithm that correlates temporal changes in expression with chromosomal position identified groups of contiguous genes expressed at discrete stages of morphological development, inferred the boundaries of known antibiotic synthesis gene loci, and revealed novel physical clusters of coordinately regulated genes. Microarray analysis of RNA from cells mutated in genes regulating synthesis of the antibiotics actinorhodin (Act) and undecylprodigiosin (Red) identified proximate and distant sites that contain putative ABC transporter and two-component system genes expressed coordinately with genes of specific biosynthetic pathways and indicated the existence of two functionally and physically discrete regulons in the Red pathway.

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Section: Discussionmentioning

confidence: 99%

Section: Microarray Analysis Of Streptomyces Transcription Genes and Dementioning

confidence: 99%

Section: Microarray Analysis Of Streptomyces Transcription Genes and Dementioning

confidence: 99%

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Global analysis of growth phase responsive gene expression and regulation of antibiotic biosynthetic pathways inStreptomyces coelicolorusing DNA microarrays

Huang¹,

Lih²,

Pan³

et al. 2001

Genes Dev.

195

188

View full text Add to dashboard Cite

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“…Plasmid pUC19 was used for the construction of cosmid subclones; plasmid pOJ260 (3) was used for monooxygenase gene disruption via Campbell-type integration. txtC was cloned into plasmid vector pET15b (Novagen) for production of a six-His [(His) 6 ]-TxtC fusion protein.…”

Section: Methodsmentioning

confidence: 99%

Involvement of a Cytochrome P450 Monooxygenase in Thaxtomin A Biosynthesis by Streptomyces acidiscabies

et al. 2002

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The biosynthesis of the thaxtomin cyclic dipeptide phytotoxins proceeds nonribosomally via the thiotemplate mechanism. Acyladenylation, thioesterification, N-methylation, and cyclization of two amino acid substrates are catalyzed by the txtAB-encoded thaxtomin synthetase. Nucleotide sequence analysis of the region 3 of txtAB in Streptomyces acidiscabies 84.104 identified an open reading frame (ORF) encoding a homolog of the P450 monooxygenase gene family. It was proposed that thaxtomin A phenylalanyl hydroxylation was catalyzed by the monooxygenase homolog. The ORF was mutated in S. acidiscabies 84.104 by using an integrative gene disruption construct, and culture filtrate extracts of the mutant were assayed for the presence of dehydroxy derivatives of thaxtomin A. Reversed-phase high-performance liquid chromatography (HPLC) and HPLC-mass spectrometry indicated that the major component in culture filtrate extracts of the mutant was less polar and smaller than thaxtomin A. Comparisons of electrospray mass spectra as well as 1 H-and 13 C-nuclear magnetic resonance spectra of the purified compound with those previously reported for thaxtomins confirmed the structure of the compound as 12,15-N-dimethylcyclo-(L-4-nitrotryptophyl-L-phenylalanyl), the didehydroxy analog of thaxtomin A. The ORF, designated txtC, was cloned and the recombinant six-His-tagged fusion protein produced in Escherichia coli and purified from cell extracts. TxtC produced in E. coli exhibited spectral properties similar to those of cytochrome P450-type hemoproteins that have undergone conversion to the catalytically inactive P420 form. Based on these properties and the high similarity of TxtC to other wellcharacterized P450 enzymes, we conclude that txtC encodes a cytochrome P450-type monooxygenase required for postcyclization hydroxylation of the cyclic dipeptide.

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“…Several lipopeptide antibiotic biosynthetic gene clusters have been isolated and partially characterized, such as fengycin (Tosato et al, 1997), mycosubtilin (Duitman et al, 1999), lichenysin (Konz et al, 1999), syringomycin (Guenzi et al, 1998), daptomycin (McHenney et al, 1998 and CDA (Chong et al, 1998;Hojati et al, 2002;Ryding et al, 2002). In addition to peptide synthase genes, genes probably involved in the synthesis of the lipid moiety of the antibiotics have been found in the gene clusters.…”

Section: Introductionmentioning

confidence: 99%

An acyl-CoA dehydrogenase is involved in the formation of the Δcis3 double bond in the acyl residue of the lipopeptide antibiotic friulimicin in Actinoplanes friuliensis

et al. 2005

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The lipopeptide antibiotic friulimicin, produced by Actinoplanes friuliensis, is an effective drug against Gram-positive bacteria, such as methicillin-resistant Staphylococcus epidermidis and Staphylococcus aureus strains. Friulimicin consists of a cyclic peptide core of ten amino acids and an acyl residue linked to an exocyclic amino acid. The acyl residue is essential for antibiotic activity, varies in length from C13 to C15, and carries a characteristic double bond at position Dcis3. Sequencing of a DNA fragment adjacent to a previously described fragment encoding some of the friulimicin biosynthetic genes revealed several genes whose gene products resemble enzymes of lipid metabolism. One of these genes, lipB, encodes an acyl-CoA dehydrogenase homologue. To elucidate the function of the LipB protein, a lipB insertion mutant was generated and the friulimicin derivative (FR242) produced by the mutant was purified. FR242 had antibiotic activity lower than friulimicin in a bioassay. Gas chromatography showed that the acyl residue of wild-type friulimicin contains a double bond, whereas a saturated bond was present in FR242. These results were confirmed by the heterologous expression of lipB in Streptomyces lividans T7, which led to the production of unsaturated fatty acids not found in the S. lividans T7 parent strain. These results indicate that the acyl-CoA dehydrogenase LipB is involved in the introduction of the unusual Dcis3 double bond into the acyl residue of friulimicin.

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Physical identification of a chromosomal locus encoding biosynthetic genes for the lipopeptide calcium-dependent antibiotic (CDA) of Streptomyces coelicolor A3(2)

Cited by 59 publications

References 28 publications

Global analysis of growth phase responsive gene expression and regulation of antibiotic biosynthetic pathways inStreptomyces coelicolorusing DNA microarrays

Global analysis of growth phase responsive gene expression and regulation of antibiotic biosynthetic pathways inStreptomyces coelicolorusing DNA microarrays

Involvement of a Cytochrome P450 Monooxygenase in Thaxtomin A Biosynthesis by Streptomyces acidiscabies

An acyl-CoA dehydrogenase is involved in the formation of the Δcis3 double bond in the acyl residue of the lipopeptide antibiotic friulimicin in Actinoplanes friuliensis

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