Isolation and structure of the novel dihydroxamate siderophore alcaligin

Nishio, Takayuki; Tanaka, Nobuo; Hiratake, Jun; Katsube, Yukiteru; Ishida, Yūzaburō; Oda, Jun’ichi

doi:10.1021/ja00234a045

Cited by 69 publications

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“…The siderophores of B. pertussis and B. bronchiseptica have been characterized structurally, and each has been identified as the macrocyclic dihydroxamate siderophore known as alcaligin (11,37), previously isolated from the taxonomically related bacterial species Alcaligenes denitrificans subsp. xylosoxydans (41,42). Characterization of B. bronchiseptica manganese-resistant mutants exhibiting deregulated expression of alcaligin and iron-repressed proteins and restoration of iron repressibility by the B. pertussis fur gene confirmed the role of the Fur repressor in regulation of these traits in both Bordetella species (9).…”

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confidence: 71%

Identification and characterization of iron-regulated Bordetella pertussis alcaligin siderophore biosynthesis genes

et al. 1996

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Bordetella bronchiseptica mutants BRM1, BRM6, and BRM9 fail to produce the native dihydroxamate siderophore alcaligin. A 4.5-kb BamHI-SmaI Bordetella pertussis genomic DNA fragment carried multiple genes required to restore alcaligin production to these siderophore-deficient mutants. Phenotypic complementation analysis using subclones of the 4.5-kb genomic region demonstrated that the closely linked BRM1 and BRM9 mutations were genetically separable from the BRM6 mutation, and both insertions exerted strong polar effects on expression of the downstream gene defined by the BRM6 mutation, suggesting a polycistronic transcriptional organization of these alcaligin biosynthesis genes. Subcloning and complementation experiments localized the putative Bordetella promoter to a 0.7-kb BamHI-SphI subregion of the cloned genomic DNA fragment. Nucleotide sequencing, phenotypic analysis of mutants, and protein expression by the 4.5-kb DNA fragment in Escherichia coli suggested the presence of three alcaligin system genes, namely, alcA, alcB, and alcC. The deduced protein products of alcA, alcB, and alcC have significant primary amino acid sequence similarities with known microbial siderophore biosynthesis enzymes. Primer extension analysis mapped the transcriptional start site of the putative alcaligin biosynthesis operon containing alcABC to a promoter region overlapping a proposed Fur repressor-binding site and demonstrated iron regulation at the transcriptional level.Iron is a fundamental nutritional requirement for virtually all cells, and its assimilation is considered essential for invading pathogenic bacteria to establish infection in the iron-limiting environment of the host (13, 56). Additionally, iron serves as an environmental modulator of the production of certain virulence factors in a number of bacteria (14,24,31,32,46). Despite host iron sequestration, mediated primarily by the glycoprotein family of iron-binding transferrins, pathogens multiply successfully in vivo because they express efficient ironscavenging systems in response to decreased iron availability. These iron retrieval systems utilize two general strategies: one involving high-affinity iron-chelating soluble siderophores (30,40) and the other using siderophore-independent cell surface receptor mechanisms allowing iron uptake directly from host sources such as transferrin, lactoferrin, and heme compounds (7,35,38,54).Bordetella pertussis, the causative agent of human whooping cough or pertussis, and Bordetella bronchiseptica, the agent of swine atrophic rhinitis and kennel cough in dogs, are bacterial pathogens that infect the respiratory epithelial mucosae of their hosts. Early reports described the production of putative siderophores by both B. pertussis and B. bronchiseptica in response to iron deficiency (1, 23). Armstrong and Clements isolated and characterized B. bronchiseptica transposon-induced siderophore-deficient mutants; DNA hybridization studies using sequences flanking those transposon insertions confirmed the existence of homologs o...

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confidence: 71%

Identification and characterization of iron-regulated Bordetella pertussis alcaligin siderophore biosynthesis genes

et al. 1996

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“…The siderophore alcaligin was first purified from the taxonomically related species A. denitrificans subsp. xylosoxydans, isolated from freshwater lake sediment (35,36). Structural analysis found alcaligin to be a symmetric 20-membered ring, 1,8(S),11,18(S)-tetrahydroxy-1,6,11,16-tetraazacycloeicosane-2,5,12,15-tetrone.…”

Section: Discussionmentioning

confidence: 99%

“…xylosoxydans (35,36). Evidence for biological activity of the purified siderophores was gathered with growth stimulation and 55 Fealcaligin transport assays of Bordetella spp.…”

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confidence: 99%

The ornithine decarboxylase gene odc is required for alcaligin siderophore biosynthesis in Bordetella spp.: putrescine is a precursor of alcaligin

Brickman

Armstrong

1996

J Bacteriol

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Chromosomal insertions defining Bordetella bronchiseptica siderophore phenotypic complementation group III mutants BRM3 and BRM5 were found to reside approximately 200 to 300 bp apart by restriction mapping of cloned genomic regions associated with the insertion markers. DNA hybridization analysis using B. bronchiseptica genomic DNA sequences flanking the cloned BRM3 insertion marker identified homologous Bordetella pertussis UT25 cosmids that complemented the siderophore biosynthesis defect of the group III B. bronchiseptica mutants. Subcloning and complementation analysis localized the complementing activity to a 2.8-kb B. pertussis genomic DNA region. Nucleotide sequencing identified an open reading frame predicted to encode a polypeptide exhibiting strong similarity at the primary amino acid level with several pyridoxal phosphate-dependent amino acid decarboxylases. Alcaligin production was fully restored to group III mutants by supplementation of iron-depleted culture media with putrescine (1,4-diaminobutane), consistent with defects in an ornithine decarboxylase activity required for alcaligin siderophore biosynthesis. Concordantly, the alcaligin biosynthesis defect of BRM3 was functionally complemented by the heterologous Escherichia coli speC gene encoding an ornithine decarboxylase activity. Enzyme assays confirmed that group III B. bronchiseptica siderophore-deficient mutants lack an ornithine decarboxylase activity required for the biosynthesis of alcaligin. Siderophore production by an analogous mutant of B. pertussis constructed by allelic exchange was undetectable. We propose the designation odc for the gene defined by these mutations that abrogate alcaligin siderophore production. Putrescine is an essential precursor of alcaligin in Bordetella spp.Nutritional iron limitation, mediated primarily by specific host iron-binding glycoproteins, is a front-line host defense against disease-causing infectious agents. Strategies aimed at defeating host iron restriction may involve the action of lowmolecular-mass, high-affinity, ferric iron-specific chelators of microbial origin, termed siderophores, that are synthesized coordinately with their cognate surface receptors and transport machinery in response to iron starvation (28). The role of siderophores in microbial pathogenesis is well established (29,53,54).Bordetella pertussis, the causative agent of human whooping cough or pertussis, and Bordetella bronchiseptica, the agent of swine atrophic rhinitis and kennel cough in dogs, are mucosal pathogens that colonize the upper respiratory tracts of their mammalian hosts. In the first reported molecular genetic studies of Bordetella iron acquisition systems, Armstrong and Clements (3) described the isolation of B. bronchiseptica mutants deficient in siderophore activity following transposon mutagenesis. DNA hybridization analysis using DNA probe sequences flanking the transposon insertions established the existence of homologs of B. bronchiseptica siderophore genes in B. pertussis. Reciprocal cross-feeding ex...

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“…The synthesis of siderophores belonging to these two main classes commonly relies on a diverse spectrum of enzymatic activities such as monooxygenases, decarboxylases, aminotransferases, ac(et)yltransferases, amino acid ligases, and aldolases (51). Siderophores synthesized by NRPSindependent pathways are found as virulence factors in several pathogens, e.g., aerobactin in enteric bacteria (114), alcaligin in B. pertussis and B. bronchiseptica (220,234), staphylobactin in Staphylococcus aureus (68), and petrobactin (formerly anthrachelin) in Bacillus anthracis (48,174). Among the NRPSindependently assembled siderophores, petrobactin represents an exception because it is capped with two catecholate moieties for iron coordination (see Fig.…”

Section: General Steps Of Siderophore Pathwaysmentioning

confidence: 99%

Siderophore-Based Iron Acquisition and Pathogen Control

Miethke

Marahiel

2007

Microbiol Mol Biol Rev

1,409

1,302

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SUMMARY High-affinity iron acquisition is mediated by siderophore-dependent pathways in the majority of pathogenic and nonpathogenic bacteria and fungi. Considerable progress has been made in characterizing and understanding mechanisms of siderophore synthesis, secretion, iron scavenging, and siderophore-delivered iron uptake and its release. The regulation of siderophore pathways reveals multilayer networks at the transcriptional and posttranscriptional levels. Due to the key role of many siderophores during virulence, coevolution led to sophisticated strategies of siderophore neutralization by mammals and (re)utilization by bacterial pathogens. Surprisingly, hosts also developed essential siderophore-based iron delivery and cell conversion pathways, which are of interest for diagnostic and therapeutic studies. In the last decades, natural and synthetic compounds have gained attention as potential therapeutics for iron-dependent treatment of infections and further diseases. Promising results for pathogen inhibition were obtained with various siderophore-antibiotic conjugates acting as “Trojan horse” toxins and siderophore pathway inhibitors. In this article, general aspects of siderophore-mediated iron acquisition, recent findings regarding iron-related pathogen-host interactions, and current strategies for iron-dependent pathogen control will be reviewed. Further concepts including the inhibition of novel siderophore pathway targets are discussed.

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Isolation and structure of the novel dihydroxamate siderophore alcaligin

Abstract: for open discussions of these results prior to publication. Supplementary Material Available: Synthetic procedures for amido complexes 1-4 and full analytical data (2 pages). Ordering information is given on any current masthead page.(30) (a) Nugent, W.

Cited by 69 publications

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Identification and characterization of iron-regulated Bordetella pertussis alcaligin siderophore biosynthesis genes

Identification and characterization of iron-regulated Bordetella pertussis alcaligin siderophore biosynthesis genes

The ornithine decarboxylase gene odc is required for alcaligin siderophore biosynthesis in Bordetella spp.: putrescine is a precursor of alcaligin

Siderophore-Based Iron Acquisition and Pathogen Control

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