Characterization of the ask?asd operon in aminoethoxyvinylglycine-producing Streptomyce s sp. NRRL 5331

Cuadrado, Yolanda; Fernández, Mónica; Recio, Eliseo; Aparicio, Jesús F.; Martı́n, Juan F.

doi:10.1007/s00253-003-1440-2

Cited by 8 publications

(4 citation statements)

References 43 publications

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“…The nested α-and β-subunits of AKs, which were originally discovered in Bacillus subtilis, 22 can be found in most Gram-positive bacteria, including mycobacteria, 33 Corynebacterium bacteria, 23,34 Amycolatopsis mediterranei 35 and Streptomyces bacteria. 36 Recent structural studies 37 have suggested that in the AK from Corynebacterium glutamicum, the nested truncated β-subunit is required for controlling feedback inhibition by the pathway endproducts threonine and lysine, possibly through structural changes in the α 2 β 2 -quaternary structure upon effector molecule binding. The catalytic AK active centre resides on the N-terminal part of the α-subunit, which is missing from the β-isoform, and it has been suggested that in the presence of threonine and lysine, the β-subunit prevents access of the substrate to the active site on the α-subunit.…”

Section: Closing Remarksmentioning

confidence: 99%

A Nested Gene in Streptomyces Bacteria Encodes a Protein Involved in Quaternary Complex Formation

Kallio

Liu

Mäntsälä

et al. 2008

Journal of Molecular Biology

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Section: Closing Remarksmentioning

confidence: 99%

A Nested Gene in Streptomyces Bacteria Encodes a Protein Involved in Quaternary Complex Formation

Kallio

Liu

Mäntsälä

et al. 2008

Journal of Molecular Biology

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“…We previously reported the identification of a 13-kb chromosomal region involved in FVG production based on the analysis of Tn5 insertion libraries in P. fluorescens WH6 (Halgren et al, 2013;Kimbrel et al, 2010). This gene cluster is distinct from those involved in the production of other oxyvinylglycines (Cuadrado et al, 2004;Rojas Murcia et al, 2015;Yasuta et al, 2001), which vary considerably despite conserved structural features of the molecules. The objectives of the current study were to: 1) systematically interrogate each gene within the 13-kb region to define the gvg cluster, and 2) investigate the transcriptional organization of the gene cluster.…”

Section: Discussionmentioning

confidence: 99%

“…Predicting the biosynthetic pathway of FVG using comparative genomics or other tools is difficult because the known biosynthetic pathways of oxyvinylglycines vary considerably (Cuadrado et al, 2004;Rojas Murcia et al, 2015;Yasuta et al, 2001) and because FVG has an unusual aminooxy linkage (Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

Functional analysis of a biosynthetic cluster essential for production of 4-formylaminooxyvinylglycine, a germination-arrest factor fromPseudomonas fluorescensWH6

Okrent

Trippe

Maselko

et al. 2016

Preprint

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Rhizosphere-associated Pseudomonas fluorescens WH6 produces the germination-arrest factor, 4-formylaminooxyvinylglycine (FVG). FVG has previously been shown to both arrest the germination of weedy grasses and to inhibit the growth of the bacterial plant pathogen Erwinia amylovora. Very little is known about the mechanism by which FVG is produced. Although a previous study identified a region of the genome that may be involved in FVG biosynthesis, it has not yet been determined which genes within that region are sufficient and necessary for FVG production. In the current study, we explored the role of each of the putative genes encoded in that region by constructing deletion mutations. Mutant strains were assayed for their ability to produce FVG with a combination of biological assays and thin-layer chromatographic analyses. This work defined the core FVG biosynthetic gene cluster and revealed several interesting characteristics of FVG production. We determined that FVG biosynthesis requires two small open reading frames of less than 150 nucleotides and that multiple transporters have overlapping but distinct functionality. In addition, two genes in the center of the biosynthetic gene cluster are not required for FVG production, suggesting that additional products may be produced from the cluster. Transcriptional analysis indicated that at least three active promoters play a role in the expression of genes within this cluster. The results of this study enrich our knowledge regarding the diversity of mechanisms by which bacteria produce non-proteinogenic amino acids like vinylglycines.

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“…The FVG analog aminoethoxyvinylglycine (AVG) is produced by Streptomyces sp. strain NRRL 5331 (33) and is structurally similar to FVG. Like FVG, AVG also inhibits the germination of grasses and is antagonistic toward E. amylovora (27,28).…”

mentioning

confidence: 98%

Resistance to Two Vinylglycine Antibiotic Analogs Is Conferred by Inactivation of Two Separate Amino Acid Transporters in Erwinia amylovora

et al. 2019

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Erwinia amylovora is the causal agent of fire blight of apple and pear trees. Several bacteria have been shown to produce antibiotics that antagonize E. amylovora, including pantocins, herbicolins, dapdiamides, and the vinylglycines, 4-formylaminooxyvinylglycine (FVG) and 4-aminoethoxyvinylglycine (AVG). Pantoea ananatis BRT175 was previously shown to exhibit antibiotic activity against E. amylovora via the production of Pantoea natural product 1 (PNP-1), later shown to be FVG; however, exposure of E. amylovora to FVG results in spontaneously resistant mutants. To identify the mechanism of resistance, we used genome variant analysis on spontaneous FVG-resistant mutants of E. amylovora and identified null mutations in the l-asparagine permease gene ansP. Heterologous expression of ansP in normally resistant Escherichia coli was sufficient to impart FVG susceptibility, suggesting that FVG is imported through this permease. Because FVG and AVG are structurally similar, we hypothesized that resistance to AVG would also be conferred through inactivation of ansP; however, ansP mutants were not resistant to AVG. We found that spontaneously resistant Ea321 mutants also arise in the presence of AVG, with whole-genome variant analysis revealing that resistance was due to inactivation of the arginine ABC transporter permease subunit gene artQ. Heterologous expression of the predicted lysE-like transporter encoded within the Pantoea ananatis BRT175 FVG biosynthetic cluster, which is likely responsible for antibiotic export, was sufficient to confer resistance to both FVG and AVG. This work highlights the important roles of amino acid transporters in antibiotic import into bacteria and the potential utility of antimicrobial amino acid analogs as antibiotics. IMPORTANCE The related antibiotics formylaminooxyvinylglycine (FVG) and aminoethoxyvinylglycine (AVG) have been shown to have activity against the fire blight pathogen Erwinia amylovora; however, E. amylovora can develop spontaneous resistance to these antibiotics. By comparing the genomes of mutants to those of the wild type, we found that inactivation of the l-asparagine transporter conferred resistance to FVG, while inactivation of the l-arginine transporter conferred resistance to AVG. We also show that the transporter encoded by the FVG biosynthetic cluster can confer resistance to both FVG and AVG. Our work indicates the important role that amino acid transporters play in the import of antibiotics and highlights the possible utility in designer antibiotics that enter the bacterial cell through amino acid transporters.

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Characterization of the ask?asd operon in aminoethoxyvinylglycine-producing Streptomyce s sp. NRRL 5331

Cited by 8 publications

References 43 publications

A Nested Gene in Streptomyces Bacteria Encodes a Protein Involved in Quaternary Complex Formation

A Nested Gene in Streptomyces Bacteria Encodes a Protein Involved in Quaternary Complex Formation

Functional analysis of a biosynthetic cluster essential for production of 4-formylaminooxyvinylglycine, a germination-arrest factor fromPseudomonas fluorescensWH6

Resistance to Two Vinylglycine Antibiotic Analogs Is Conferred by Inactivation of Two Separate Amino Acid Transporters in Erwinia amylovora

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