Expression of multiple pili by Legionella gneumophila:: Identification and characterization of a type IV pilin gene and its role in adherence to mammalian and protozoan cells

Legionella pneumophila expresses pill of variable lengths, either long (0.8 to 1.5 mu m) or short (0.1 to 0.6 mu m), that can be observed by transmission electron microscopy. We have identified a gene in L. pneumophila with homology to the type IV pilin genes (pilE(L)). An insertion mutation was constructed in pilE(L) and introduced into the L. pneumophila wild-type strain by allelic exchange. The pilin mutant is defective for expression of long pill. Reintroduction of the pilin locus on a cosmid vector restores expression of the long pill. The L. pneumophila pilE(L) mutant exhibited approximately a 50% decrease in adherence to human epithelial cells (HeLa and WI-26 cells), macrophages (U937 cells), and Acanthamoeba polyphaga but had a wild-type phenotype for intracellular replication within these cells. Southern hybridization analysis shelved that the pilE(L) locus is present in L. pneumophila serogroups 1 through 13 but is variable in 16 other Legionella species. The presence of a type TV pilin gene and its expression by L. pneumophila may provide an advantage for colonization of lung tissues during Legionnaires' disease and invasion of amoebas in the environment.