Phagocyte meets prey: Uptake, internalization, and killing of bacteria by Dictyostelium amoebae

Abstract

Dictyostelium cells are professional phagocytes that avidly consume bacteria, their natural prey. Fluorescent probes have allowed us to monitor the initial steps in this process in living cells. Using probes that bind to F-actin, we have visualized the assembly and disassembly of actin filaments responsible for extending the phagocytic cup to engulf a bacterium, and, after the phagosome has sealed, the assembly of new actin filaments to propel the phagosome away from the site of uptake. Using bacteria expressing fluorescent proteins that are susceptible to proteolysis, we have monitored the loss of that fluorescent signal and the staining of the bacterial contents with neutral red, indicating permeabilization of the bacterial cell wall and acidification of the cytoplasm. We find that acidification occurs during a period of microtubule-based transport that promotes fusion of the phagosome with microtubule-associated acidic endosomes. Actin-powered phagosome internalization, transport of the phagosome along microtubules, proteolysis and acidification of bacterial contents, all typically occur within the first six or seven minutes after formation of the phagosome. Thus, tracking individual phagosomes has revealed that early steps in phagosome maturation occur much more rapidly than had been inferred from previous population studies.