Dynamic properties of Legionella-containing phagosomes in Dictyostelium amoebae.
The natural hosts of the bacterial pathogen Legionella pneumophila are amoebae and protozoa. In these hosts, as in human macrophages, the pathogen enters the cell through phagocytosis, then rapidly modifies the phagosome to create a compartment that supports its replication. We have examined L. pneumophila entry and behaviour during early stages of the infection of Dictyostelium discoideum amoebae. Bacteria were labelled with a red fluorescent marker, and selected proteins and organelles in the host were labelled with GFP, allowing the dynamics and interactions of L. pneumophila -containing phagosomes to be tracked in living cells. These studies demonstrated that entry of L. pneumophila is an actin-mediated process, that the actin-binding protein coronin surrounds the nascent phagosome but dissociates immediately after internalization, that ER membrane is not incorporated into a phagosome during uptake, that the newly internalized phagosome is rapidly transported about the cell on microtubules, that association of ER markers with the phagosome occurs in two steps that correlate with distinct changes in phagosome movement, and that the vacuolar H(+)-ATPase does not associate with mature replication vacuoles. These studies have clarified certain aspects of the infection process and provided new insights into the dynamic interactions between the pathogen and its host.
Lu
Hao
H
Program in Molecular, Cell, and Developmental Biology, Oklahoma Medical Research Foundation, Oklahoma City, OK 73121, USA.
Clarke
Margaret
M
D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000032:analysis; D000199:Actins / Q000378:metabolism; D000818:Animals; D004023:Dictyostelium / Q000382:microbiology* / Q000648:ultrastructure; D004721:Endoplasmic Reticulum / Q000378:metabolism / Q000382:microbiology; D005453:Fluorescence; D007425:Intracellular Membranes / Q000378:metabolism / Q000382:microbiology; D016952:Legionella pneumophila / Q000502:physiology*; D018613:Microscopy, Confocal; D008870:Microtubules / Q000382:microbiology; D010588:Phagosomes / Q000382:microbiology*; D006180:Proton-Translocating ATPases / Q000378:metabolism; D013194:Staining and Labeling; D054378:Acetogenins
D015107:4-Butyrolactone; D000199:Actins; D006180:Proton-Translocating ATPases; C438920:coronin acetogenin; D054378:Acetogenins