title: Investigating the Role of the Host Multidrug Resistance Associated Protein Transporter Family in Burkholderia cepacia Complex Pathogenicity Using a Caenorhabditis elegans Infection Model.
abstract: This study investigated the relationship between host efflux system of the non-vertebrate nematode Caenorhabditis elegans and Burkholderia cepacia complex (Bcc) strain virulence. This is the first comprehensive effort to profile host-transporters within the context of Bcc infection. With this aim, two different toxicity tests were performed: a slow killing assay that monitors mortality of the host by intestinal colonization and a fast killing assay that assesses production of toxins. A Virulence Ranking scheme was defined, that expressed the toxicity of the Bcc panel members, based on the percentage of surviving worms. According to this ranking the 18 Bcc strains were divided in 4 distinct groups. Only the Cystic Fibrosis isolated strains possessed profound nematode killing ability to accumulate in worms' intestines. For the transporter analysis a complete set of isogenic nematode single Multidrug Resistance associated Protein (MRP) efflux mutants and a number of efflux inhibitors were interrogated in the host toxicity assays. The Bcc pathogenicity profile of the 7 isogenic C. elegans MRP knock-out strains functionality was classified in two distinct groups. Disabling host transporters enhanced nematode mortality more than 50% in 5 out of 7 mutants when compared to wild type. In particular mrp-2 was the most susceptible phenotype with increased mortality for 13 out 18 Bcc strains, whereas mrp-3 and mrp-4 knock-outs had lower mortality rates, suggesting a different role in toxin-substrate recognition. The use of MRP efflux inhibitors in the assays resulted in substantially increased (>40% on average) mortality of wild-type worms.
authors.affiliation: Institute of Protein Biochemistry, National Research Council, Via P. Castellino 111, I-80131, Naples, Italy.|Department of Chemical Sciences and School of Biotechnological Sciences, University of Naples Federico II, Via Cintia, I-80126, Naples, Italy.
authors.forename: Pietro
authors.initials: P
authors.lastname: Tedesco
authors.affiliation: Institute of Protein Biochemistry, National Research Council, Via P. Castellino 111, I-80131, Naples, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Visone
authors.affiliation: Department of Chemical Sciences and School of Biotechnological Sciences, University of Naples Federico II, Via Cintia, I-80126, Naples, Italy.
authors.forename: Ermenegilda
authors.initials: E
authors.lastname: Parrilli
authors.affiliation: Department of Chemical Sciences and School of Biotechnological Sciences, University of Naples Federico II, Via Cintia, I-80126, Naples, Italy.
authors.forename: Maria Luisa
authors.initials: ML
authors.lastname: Tutino
authors.affiliation: Laboratory of Microbial and Molecular Evolution, Department of Biology, University of Florence, Via Madonna del Piano, I-50019, Sesto Fiorentino, Florence, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Perrin
authors.affiliation: Laboratory of Microbial and Molecular Evolution, Department of Biology, University of Florence, Via Madonna del Piano, I-50019, Sesto Fiorentino, Florence, Italy.
authors.forename: Isabel
authors.initials: I
authors.lastname: Maida
authors.affiliation: Laboratory of Microbial and Molecular Evolution, Department of Biology, University of Florence, Via Madonna del Piano, I-50019, Sesto Fiorentino, Florence, Italy.
authors.forename: Renato
authors.initials: R
authors.lastname: Fani
authors.affiliation: School of Biotechnology and Biomolecular Sciences and Centre for Marine Bio-Innovation, University of New South Wales, Sydney, 2052, New South Wales, Australia.
authors.forename: Francesco
authors.initials: F
authors.lastname: Ballestriero
authors.affiliation: Torrey Pines Institute of Molecular Studies, Port St. Lucie, FL, United States of America, and San Diego, CA, United States of America.
authors.forename: Radleigh
authors.initials: R
authors.lastname: Santos
authors.affiliation: Torrey Pines Institute of Molecular Studies, Port St. Lucie, FL, United States of America, and San Diego, CA, United States of America.
authors.forename: Clemencia
authors.initials: C
authors.lastname: Pinilla
authors.affiliation: Institute of Bioscience and BioResources, National Research Council, via P. Castellino 111, I-80131, Naples, Italy.|Institute of Genetics and Biophysics, National Research Council, via P. Castellino 111, I-80131, Naples, Italy.
authors.forename: Elia
authors.initials: E
authors.lastname: Di Schiavi
authors.affiliation: Torrey Pines Institute of Molecular Studies, Port St. Lucie, FL, United States of America, and San Diego, CA, United States of America.|Wellman Centre for Photomedicine, Massachusetts General Hospital, Boston, MA, United States of America.|Department of Dermatology, Harvard Medical School, Boston, MA, United States of America.
authors.forename: George
authors.initials: G
authors.lastname: Tegos
authors.affiliation: Institute of Protein Biochemistry, National Research Council, Via P. Castellino 111, I-80131, Naples, Italy.
authors.forename: Donatella
authors.initials: D
authors.lastname: de Pascale
mesh_terms: D000818:Animals; D019121:Burkholderia Infections / Q000188:drug therapy / Q000235:genetics* / Q000382:microbiology; D016956:Burkholderia cepacia / Q000187:drug effects* / Q000472:pathogenicity; D017173:Caenorhabditis elegans / Q000187:drug effects / Q000235:genetics; D004195:Disease Models, Animal; D018432:Drug Resistance, Multiple / Q000235:genetics*; D055786:Gene Knockout Techniques; D006801:Humans; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C000715087:ABCC2 protein, human