title: Iron acquisition functions expressed by the human pathogen Acinetobacter baumannii.
abstract: Acinetobacter baumannii is a gram-negative bacterium that causes serious infections in compromised patients. More recently, it has emerged as the causative agent of severe infections in military personnel wounded in Iraq and Afghanistan. This pathogen grows under a wide range of conditions including iron-limiting conditions imposed by natural and synthetic iron chelators. Initial studies using the type strain 19606 showed that the iron proficiency of this pathogen depends on the expression of the acinetobactin-mediated iron acquisition system. More recently, we have observed that hemin but not human hemoglobin serves as an iron source when 19606 isogenic derivatives affected in acinetobactin transport and biosynthesis were cultured under iron-limiting conditions. This finding is in agreement with the observation that the genome of the strain 17978 has a gene cluster coding for putative hemin-acquisition functions, which include genes coding for putative hemin utilization functions and a TonBExbBD energy transducing system. This system restored enterobactin biosynthesis in an E. coli ExbBD deficient strain but not when introduced into a TonB mutant. PCR and Southern blot analyses showed that this hemin-utilization gene cluster is also present in the 19606 strain. Analysis of the 17978 genome also showed that this strain harbors genes required for acinetobactin synthesis and transport as well as a gene cluster that could code for additional iron acquisition functions. This hypothesis is in agreement with the fact that the inactivation of the basD acinetobactin biosynthetic gene did not affect the growth of A. baumannii 17978 cells under iron-chelated conditions. Interestingly, this second iron uptake gene cluster is flanked by perfect inverted repeats and includes transposase genes that are expressed transcriptionally. Also interesting is the observation that this additional cluster could not be detected in the type strain 19606, an observation that suggests some significant differences in the iron uptake capacity between these two A. baumannii strains. Transposome mutagenesis of the strain 19606 resulted in the isolation of a derivative unable to grow under iron-chelated conditions. Gene mapping and protein analysis together with complementation assays showed that a protein related to SecA, which is a component of the Sec protein secretion system in a wide range of bacteria, is needed at least for the production of the BauA acinetobactin outer membrane receptor. Furthermore, this derivative was unable to use hemin as an iron source under limiting conditions. Taken together, these results indicate that A. baumannii expresses siderophore-mediated and hemin acquisition functions, although different isolates differ in their iron acquisition capacity. Unexpectedly, the ability of this pathogen to acquire iron depends on the expression of a SecA protein secretion function, which has not been associated with iron acquisition in bacteria.
authors.affiliation: Department of Microbiology, Miami University, Oxford, OH 45056, USA.
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Zimbler
authors.forename: William F
authors.initials: WF
authors.lastname: Penwell
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Gaddy
authors.forename: Sharon M
authors.initials: SM
authors.lastname: Menke
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Tomaras
authors.forename: Pamela L
authors.initials: PL
authors.lastname: Connerly
authors.forename: Luis A
authors.initials: LA
authors.lastname: Actis
mesh_terms: D040981:Acinetobacter baumannii / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity; D005708:Gallium / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006427:Hemin / Q000235:genetics / Q000378:metabolism; D006454:Hemoglobins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007501:Iron / Q000378:metabolism*; D005810:Multigene Family; D017262:Siderophores / Q000235:genetics / Q000378:metabolism
chemical_list: D006454:Hemoglobins; D017262:Siderophores; D006427:Hemin; D005708:Gallium; D007501:Iron