title: Genes from pUM505 plasmid contribute to Pseudomonas aeruginosa virulence.
abstract: The pUM505 plasmid was isolated from a clinical strain of Pseudomonas aeruginosa. This plasmid contains a genomic island with sequence similar to islands found in chromosomes of virulent P. aeruginosa clinical isolates. The objective of this work was to determine whether pUM505 increases the virulence of P. aeruginosa and to identify the genes responsible for this property. First, using the lettuce-leaf model, we found that pUM505 significantly increases the virulence of P. aeruginosa reference strain PAO1. pUM505 also increased the PAO1 virulence in a murine model and increased cytotoxicity of this strain toward HeLa cells. Thus, we generated a pUM505 gene library of 103 clones in the pUCP20 binary vector. The library was transferred to Escherichia coli TOP10 and P. aeruginosa PAO1 to identify genes. The lettuce-leaf model allowed us to identify three recombinant plasmids that increased the virulence of both E. coli and P. aeruginosa strains. These recombinant plasmids also increased the virulence of the PAO1 strain in mice and induced a cytotoxic effect in HeLa cells. Eleven genes were identified in the virulent transformants. Of these genes, only the pUM505 ORF 2 has homology with a gene previously implicated in virulence. These results indicate that pUM505 contains several genes that encode virulence factors, suggesting that the plasmid may contribute directly to bacterial virulence.
authors.affiliation: Instituto de Investigaciones Químico-Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Ciudad Universitaria, Edificio B-3, 58030, Morelia, Mich, México.
authors.forename: E
authors.initials: E
authors.lastname: Rodríguez-Andrade
authors.affiliation: Instituto de Investigaciones Químico-Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Ciudad Universitaria, Edificio B-3, 58030, Morelia, Mich, México.
authors.forename: K C
authors.initials: KC
authors.lastname: Hernández-Ramírez
authors.affiliation: Instituto de Investigaciones Químico-Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Ciudad Universitaria, Edificio B-3, 58030, Morelia, Mich, México.
authors.forename: S P
authors.initials: SP
authors.lastname: Díaz-Peréz
authors.affiliation: Instituto de Investigaciones Químico-Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Ciudad Universitaria, Edificio B-3, 58030, Morelia, Mich, México.
authors.forename: A
authors.initials: A
authors.lastname: Díaz-Magaña
authors.affiliation: Instituto de Investigaciones Químico-Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Ciudad Universitaria, Edificio B-3, 58030, Morelia, Mich, México.
authors.forename: M P
authors.initials: MP
authors.lastname: Chávez-Moctezuma
authors.affiliation: Instituto de Investigaciones Químico-Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Ciudad Universitaria, Edificio B-3, 58030, Morelia, Mich, México.
authors.forename: V
authors.initials: V
authors.lastname: Meza-Carmen
authors.affiliation: Facultad de Químico- Farmacobiología, Universidad Michoacana de San Nicolás de Hidalgo, Morelia, México.
authors.forename: R
authors.initials: R
authors.lastname: Ortíz-Alvarado
authors.affiliation: Instituto de Investigaciones Químico-Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Ciudad Universitaria, Edificio B-3, 58030, Morelia, Mich, México.
authors.forename: C
authors.initials: C
authors.lastname: Cervantes
authors.affiliation: Instituto de Investigaciones Químico-Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Ciudad Universitaria, Edificio B-3, 58030, Morelia, Mich, México. marthaisela_ramirez@hotmail.com.
authors.forename: M I
authors.initials: MI
authors.lastname: Ramírez-Díaz
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D004195:Disease Models, Animal; D015723:Gene Library; D005798:Genes, Bacterial; D006801:Humans; D018545:Lactuca / Q000382:microbiology; D008297:Male; D051379:Mice; D010935:Plant Diseases / Q000382:microbiology; D018515:Plant Leaves / Q000382:microbiology; D010957:Plasmids / Q000235:genetics*; D011552:Pseudomonas Infections / Q000382:microbiology / Q000401:mortality; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000472:pathogenicity*; D014774:Virulence / Q000235:genetics
keywords: Citotoxicity; Genomic island; Human cells; Mouse; Pathogenicity