1887

Abstract

is an opportunistic pathogen causing serious infections in patients with cystic fibrosis. The widespread distribution of this bacterium in the environment suggests that it must adapt to stress to be able to survive. We identified in K56-2 a gene predicted to encode RpoE, the extra-cytoplasmic stress response regulator. The gene is the first gene of a predicted operon encoding proteins homologous to RseA, RseB, MucD and a protein of unknown function. The genomic organization and the co-transcription of these genes were confirmed by PCR and RT-PCR. The and genes were mutated, giving rise to RSF24 and RSF25, respectively. While mutant RSF24 did not demonstrate any growth defects under the conditions tested, RSF25 was compromised for growth under temperature (44 °C) and osmotic stress (426 mM NaCl). Expression of RpoE could complement the osmotic growth defect but exacerbated temperature sensitivity in both RSF25 and wild-type K56-2. Inactivation of altered the bacterial cell surface, as indicated by increased binding of the fluorescent dye calcofluor white and by an altered outer-membrane protein profile. These cell surface changes were restored by complementation with a plasmid encoding . Macrophage infections in which bacterial colocalization with fluorescent dextran was examined demonstrated that the mutant could not delay the fusion of containing vacuoles with lysosomes, in contrast to the parental strain K56-2. These data show that is required for bacterial growth under certain stress conditions and for the ability of intracellular bacteria to delay phagolysosomal fusion in macrophages.

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2008-02-01
2019-10-17
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