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1.
Cell Microbiol ; 11(7): 1128-50, 2009 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-19388904

RESUMEN

Summary The highly infectious bacterium Francisella tularensis is a facultative intracellular pathogen, whose virulence requires proliferation inside host cells, including macrophages. Here we have performed a global transcriptional profiling of the highly virulent F. tularensis ssp. tularensis Schu S4 strain during its intracellular cycle within primary murine macrophages, to characterize its intracellular biology and identify pathogenic determinants based on their intracellular expression profiles. Phagocytosed bacteria rapidly responded to their intracellular environment and subsequently altered their transcriptional profile. Differential gene expression profiles were revealed that correlated with specific intracellular locale of the bacteria. Upregulation of general and oxidative stress response genes was a hallmark of the early phagosomal and late endosomal stages, while induction of transport and metabolic genes characterized the cytosolic replication stage. Expression of the Francisella Pathogenicity Island (FPI) genes, which are required for intracellular proliferation, increased during the intracellular cycle. Similarly, 27 chromosomal loci encoding putative hypothetical, secreted, outer membrane proteins or transcriptional regulators were identified as upregulated. Among these, deletion of FTT0383, FTT0369c or FTT1676 abolished the ability of Schu S4 to survive or proliferate intracellularly and cause lethality in mice, therefore identifying novel determinants of Francisella virulence from their intracellular expression profile.


Asunto(s)
Francisella tularensis/fisiología , Perfilación de la Expresión Génica , Macrófagos/microbiología , Análisis de Secuencia por Matrices de Oligonucleótidos , Factores de Virulencia/biosíntesis , Animales , Transporte Biológico , Células Cultivadas , Citosol/microbiología , Endosomas/microbiología , Francisella tularensis/crecimiento & desarrollo , Francisella tularensis/patogenicidad , Genes Bacterianos , Islas Genómicas , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Estrés Oxidativo , Fagosomas/microbiología , Estrés Fisiológico , Virulencia
2.
Infect Immun ; 76(6): 2273-83, 2008 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-18347045

RESUMEN

Chlamydia trachomatis possesses a cryptic 7.5-kb plasmid of unknown function. Here, we describe a comprehensive molecular and biological characterization of the naturally occurring plasmidless human C. trachomatis strain L2(25667R). We found that despite minimal chromosomal polymorphisms, the LGV strain L2(25667R) was indistinguishable from plasmid-positive strain L2(434) with regard to its in vitro infectivity characteristics such as growth kinetics, plaquing efficiency, and plaque size. The only in vitro phenotypic differences between L2(434) and L2(25667R) were the accumulation of glycogen granules in the inclusion matrix and the lack of the typical intrainclusion Brownian-like movement characteristic of C. trachomatis strains. Conversely, we observed a marked difference between the two strains in their abilities to colonize and infect the female mouse genital tract. The 50% infective dose of plasmidless strain L2(25667R) was 400-fold greater (4 x 10(6) inclusion-forming units [IFU]) than that of plasmid-bearing strain L2(434) (1 x 10(4) IFU). Transcriptome analysis of the two strains demonstrated a decrease in the transcript levels of a subset of chromosomal genes for strain L2(25667R). Among those genes was glgA, encoding glycogen synthase, a finding consistent with the failure of L2(25667R) to accumulate glycogen granules. These findings support a primary role for the plasmid in in vivo infectivity and suggest that virulence is controlled, at least in part, by the plasmid's ability to regulate the expression of chromosomal genes. Our findings have important implications in understanding a role for the plasmid in the pathogenesis of human infection and disease.


Asunto(s)
Chlamydia trachomatis/fisiología , Cromosomas Bacterianos/genética , Plásmidos/fisiología , Transcripción Genética/fisiología , Factores de Virulencia/fisiología , Animales , Técnicas Bacteriológicas , Infecciones por Chlamydia/microbiología , Chlamydia trachomatis/clasificación , Chlamydia trachomatis/citología , Chlamydia trachomatis/genética , Femenino , Perfilación de la Expresión Génica , Regulación Bacteriana de la Expresión Génica/fisiología , Ratones , Ratones Endogámicos C3H , Plásmidos/genética , Polimorfismo Genético , Análisis por Matrices de Proteínas , Vaginosis Bacteriana/microbiología , Factores de Virulencia/genética
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