The Legionella pneumophila GacA homolog (LetA) is involved in the regulation of icm virulence genes and is required for intracellular multiplication in Acanthamoeba castellanii

Abstract

Legionella pneumophila, the causative agent of legionnaires' disease, is a broad-host-range facultative intracellular pathogen. Thus far, 24 genes ( icm/ dot genes) required for L. pneumophila intracellular growth, have been discovered. In this study, a deletion substitution was constructed in the L. pneumophila homolog of the gacA response regulator ( letA) and its involvement in L. pneumophila pathogenicity and icm/ dot gene expression was characterized. The letA mutant constructed had no intracellular growth defect when analyzed in HL-60 derived human macrophages, but it was found to be severely attenuated for intracellular growth in the protozoan host Acanthamoeba castellanii. The growth defect in amoebae was fully complemented by introducing the L. pneumophila letA gene on a plasmid. In addition, the LetA regulator was found to be involved in the expression of three icm genes ( icmT, icmP and icmR). The level of expression of the icmT::lacZ and icmR::lacZ fusions was found to be higher, while the level of expression of the icmP::lacZ fusion was found to be lower when analyzed in the letA mutant strain, in comparison to the wild-type strain. We concluded that LetA has a moderate effect on icm/ dot gene expression, but it probably plays a major role in the expression of L. pneumophila genes required for intracellular growth in protozoan hosts. A similar host specific phenotype was previously described for the RpoS sigma factor and the type II general secretion system of L. pneumophila.