Biosynthesis of the dideoxysugar component of jadomycin B: genes in the jad cluster of Streptomyces venezuelae ISP5230 for l-digitoxose assembly and transfer to the angucycline aglycone
               The GenBank accession number for the sequence reported in this paper is AY026363.

Wang, Liru; White, Robert L.; Vining, L. C.

doi:10.1099/00221287-148-4-1091

Cited by 51 publications

(57 citation statements)

References 68 publications

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“…Because cmlK is located 55 bp upstream of cmlS, and the two genes are transcribed in the same direction, loss of halogenase activity might be attributed to a polar effect of the cmlK disruption preventing expression of cmlS. However, three observations argue against this: (i) the DNA fragment cloned in pJV506 contains a putative transcriptional terminator, consisting of multiple repeat sequences capable of generating hairpin loops, in the region between cmlK and cmlS; (ii) the Am R cassette used to disrupt cmlK contains a promoter from which the resistance marker and downstream genes with the same transcriptional orientation are expressed; the cassette lacks a terminator and has been shown (Wang et al, 2002) to support transcription of genes downstream of the insertion site when it is introduced in the appropriate orientation into polycistronic operons. The Am R gene in the insertionally inactivated cmlK mutant was shown by restriction analyses to have the same transcriptional orientation as cmlS; (iii) when a 3?57 kb EcoRI fragment of pJV502 containing the complete cmlK sequence as well as 2 kb of DNA upstream of the start codon was cloned in the conjugal vector pJV528, and transferred conjugally into S. venezuelae VS1111 to complement in trans the chromosomal cmlK mutation in that strain, selection of transconjugants (VS1115) with an Am R Ts R phenotype yielded cultures in which Cm production was partially restored.…”

Section: Orf11 (Encoding Cmlk An Amp-ligase)mentioning

confidence: 94%

Biosynthesis of the dichloroacetyl component of chloramphenicol in Streptomyces venezuelae ISP5230: genes required for halogenation

2004

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Five ORFs were detected in a fragment from the Streptomyces venezuelae ISP5230 genomic DNA library by hybridization with a PCR product amplified from primers representing a consensus of known halogenase sequences. Sequencing and functional analyses demonstrated that ORFs 11 and 12 (but not ORFs 13-15) extended the partially characterized gene cluster for chloramphenicol (Cm) biosynthesis in the chromosome. Disruption of ORF11 (cmlK) or ORF12 (cmlS) and conjugal transfer of the insertionally inactivated genes to S. venezuelae gave mutant strains VS1111 and VS1112, each producing a similar series of Cm analogues in which unhalogenated acyl groups replaced the dichloroacetyl substituent of Cm. 1 H-NMR established that the principal metabolite in the disrupted strains was the a-N-propionyl analogue. The sequence of CmlK implicated the protein in adenylation, and involvement in halogenation was inferred from biosynthesis of analogues by the cmlK-disrupted mutant. A role in generating the dichloroacetyl substituent was supported by partial restoration of Cm biosynthesis when a cloned copy of cmlK was introduced in trans into VS1111. Complementation of the mutant also indicated that inactivation of cmlK rather than a polar effect of the disruption on cmlS expression had interfered with dichloroacetyl biosynthesis. The deduced CmlS sequence resembled sequences of FADH 2 -dependent halogenases. Conjugal transfer of cmlK or cmlS into S. venezuelae cml-2, a chlorination-deficient strain with a mutation mapped genetically to the Cm biosynthesis gene cluster, did not complement the cml-2 lesion, suggesting that one or more genes in addition to cmlK and cmlS is needed to assemble the dichloroacetyl substituent. Insertional inactivation of ORF13 did not affect Cm production, and the products of ORF14 and ORF15 matched Streptomyces coelicolor A3(2) proteins lacking plausible functions in Cm biosynthesis. Thus cmlS appears to mark the downstream end of the gene cluster.

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Section: Orf11 (Encoding Cmlk An Amp-ligase)mentioning

confidence: 94%

Biosynthesis of the dichloroacetyl component of chloramphenicol in Streptomyces venezuelae ISP5230: genes required for halogenation

2004

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“…To test the importance of jadX for production of Jd, comparative Jd production growths were conducted for S. venezuelae ISP5230 VS1085 (WT) and S. venezuelae VS1085 mutant (ΔjadX::aac(3)IV), a mutant strain in which jadX is disrupted via insertion of an apramycin-resistance cassette. 14 Our analysis of other apramycin-blocked mutants in the dideoxysugar pathway resulted in the characterization of metabolites not initially identified in previous studies, including ad i fferentially glycosylated jadomycin. 27 We therefore reconfirmed by PCR the location of the apramycin-resistance cassette in the jadomycin biosynthetic cluster for the ΔjadX::aac(3)IV mutant.…”

Section: ■ Introductionmentioning

confidence: 97%

JadX is a Disparate Natural Product Binding Protein

Robertson¹,

Forget²,

Martinez-Farina³

et al. 2016

J. Am. Chem. Soc.

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Vous avez des questions? Nous pouvons vous aider. Pour communiquer directement avec un auteur, consultez la première page de la revue dans laquelle son article a été publié afin de trouver ses coordonnées. Si vous n'arrivez pas à les repérer, communiquez avec nous à PublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. Questions? Contact the NRC Publications Archive team atPublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. If you wish to email the authors directly, please see the first page of the publication for their contact information. NRC Publications Archive Archives des publications du CNRCThis publication could be one of several versions: author's original, accepted manuscript or the publisher's version. / La version de cette publication peut être l'une des suivantes : la version prépublication de l'auteur, la version acceptée du manuscrit ou la version de l'éditeur. NRC Publications Record / Notice d'Archives des publications de CNRC:http://nparc.cisti-icist.nrc-cnrc.gc.ca/eng/view/object/?id=1cef32c4-4c83-4b35-af83-2602ff6d1af3 http://nparc.cisti-icist.nrc-cnrc.gc.ca/fra/voir/objet/?id=1cef32c4-4c83-4b35-af83-2602ff6d1af3JadX is a Disparate Natural Product Binding Protein ABSTRACT: We report that JadX, a protein of previously undetermined function coded for in the jadomycin biosynthetic gene cluster of Streptomyces venezuelae ISP5230, affects both chloramphenicol and jadomycin production levels in blocked mutants. Characterization of recombinant JadX through protein−ligand interactions by chemical shift perturbation and WaterLOGSY NMR spectroscopy resulted in the observation of binding between JadX and a series of jadomycins and between JadX and chloramphenicol, another natural product produced by S. venezuelae ISP5230. These results suggest JadX to be an unusual class of natural product binding protein involved in binding structurally disparate natural products. The ability for JadX to bind two different natural products in vitro and the ability to affect production of these secondary metabolites in vivo suggest a potential role in regulation or signaling. This is the first example of functional characterization of these JadX-like proteins, and provides insight into a previously unobserved regulatory process.

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“…S12) (39). The putative 4-Omethyldigitoxose biosynthesis proteins are homologous to a similar suite of proteins responsible for digitoxose biosynthesis in the BGC for jadomycin B in Streptomyces venezuelae ISP5230 (40). However, the selvamicin sugar subcluster contains an additional O-methyltransferase gene (selSI) and lacks an NDP-sugar 4-ketoreductase, which would normally be required for digitoxose formation.…”

Section: Chemistrymentioning

confidence: 99%

Selvamicin, an atypical antifungal polyene from two alternative genomic contexts

Arnam

Ruzzini

Sit

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

100

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The bacteria harbored by fungus-growing ants produce a variety of small molecules that help maintain a complex multilateral symbiosis. In a survey of antifungal compounds from these bacteria, we discovered selvamicin, an unusual antifungal polyene macrolide, in bacterial isolates from two neighboring ant nests. Selvamicin resembles the clinically important antifungals nystatin A1 and amphotericin B, but it has several distinctive structural features: a noncationic 6-deoxymannose sugar at the canonical glycosylation site and a second sugar, an unusual 4-O-methyldigitoxose, at the opposite end of selvamicin’s shortened polyene macrolide. It also lacks some of the pharmacokinetic liabilities of the clinical agents and appears to have a different target. Whole genome sequencing revealed the putative type I polyketide gene cluster responsible for selvamicin’s biosynthesis including a subcluster of genes consistent with selvamicin’s 4-O-methyldigitoxose sugar. Although the selvamicin biosynthetic cluster is virtually identical in both bacterial producers, in one it is on the chromosome, in the other it is on a plasmid. These alternative genomic contexts illustrate the biosynthetic gene cluster mobility that underlies the diversity and distribution of chemical defenses by the specialized bacteria in this multilateral symbiosis.

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Biosynthesis of the dideoxysugar component of jadomycin B: genes in the jad cluster of Streptomyces venezuelae ISP5230 for l-digitoxose assembly and transfer to the angucycline aglycone The GenBank accession number for the sequence reported in this paper is AY026363.

Cited by 51 publications

References 68 publications

Biosynthesis of the dichloroacetyl component of chloramphenicol in Streptomyces venezuelae ISP5230: genes required for halogenation

Biosynthesis of the dichloroacetyl component of chloramphenicol in Streptomyces venezuelae ISP5230: genes required for halogenation

JadX is a Disparate Natural Product Binding Protein

Selvamicin, an atypical antifungal polyene from two alternative genomic contexts

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