Identification and transcriptional organization of a gene cluster involved in biosynthesis and transport of acinetobactin, a siderophore produced by Acinetobacter baumannii ATCC 19606T

Kazutoshi Mihara, Tomotaka Tanabe, Yoshiko Yamakawa, Tatsuya Funahashi, Hiroshi Nakao, Shizuo Narimatsu, Shigeo Yamamoto

Research output: Contribution to journalArticle

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Abstract

In order to assimilate iron, Acinetobacter baumannii ATCC 19606T produces a siderophore named acinetobactin (Ab) that is composed of equimolar quantities of 2,3-dihydroxybenzoic acid (DHBA), L-threonine and N-hydroxyhistamine. Application of the Fur titration assay system to A. baumannii genomic libraries, followed by further cloning of the regions surrounding the candidate genes, led to the identification of the Ab cluster, which harbours the genetic determinants necessary for the biosynthesis and transport of the siderophore. However, an entA homologue essential for DHBA biosynthesis was not found in this cluster. Functions of potential biosynthetic genes inferred by homology studies suggested that the precursors, DHBA, L-threonine and N-hydroxyhistamine, are linked in steps resembling those of bacterial non-ribosomal peptide synthesis to form Ab. Genes responsible for the two-step biosynthesis of N-hydroxyhistamine from histidine were also identified in this cluster. Their genetic organization suggests that five genes involved in the transport system of ferric Ab into the cell cytosol form an operon. Construction of disruptants of some selected genes followed by phenotypic analysis supported their predicted biological functions. Interestingly, three additional genes probably involved in the intracellular release of iron from ferric Ab and the secretion of nascent Ab are contained in this cluster. Primer extension and RT-PCR analyses suggested that the Ab cluster, which includes 18 genes, is organized in seven transcriptional units originating from respective Fur-regulated promoter-operator regions.

Original languageEnglish
Pages (from-to)2587-2597
Number of pages11
JournalMicrobiology
Volume150
Issue number8
Publication statusPublished - Aug 2004

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Siderophores
Acinetobacter baumannii
Multigene Family
Genes
Threonine
Iron
Acids
Genomic Library
Operon
acinetobactin
Histidine
Genetic Promoter Regions
Cytosol
Organism Cloning
Polymerase Chain Reaction
Peptides

ASJC Scopus subject areas

  • Microbiology

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Identification and transcriptional organization of a gene cluster involved in biosynthesis and transport of acinetobactin, a siderophore produced by Acinetobacter baumannii ATCC 19606T. / Mihara, Kazutoshi; Tanabe, Tomotaka; Yamakawa, Yoshiko; Funahashi, Tatsuya; Nakao, Hiroshi; Narimatsu, Shizuo; Yamamoto, Shigeo.

In: Microbiology, Vol. 150, No. 8, 08.2004, p. 2587-2597.

Research output: Contribution to journalArticle

Mihara, Kazutoshi ; Tanabe, Tomotaka ; Yamakawa, Yoshiko ; Funahashi, Tatsuya ; Nakao, Hiroshi ; Narimatsu, Shizuo ; Yamamoto, Shigeo. / Identification and transcriptional organization of a gene cluster involved in biosynthesis and transport of acinetobactin, a siderophore produced by Acinetobacter baumannii ATCC 19606T. In: Microbiology. 2004 ; Vol. 150, No. 8. pp. 2587-2597.
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