Inhibition of early endosome fusion by Trypanosoma cruzi-infected macrophage cytosol.
Trypanosoma cruzi trypomastigotes survive inside macrophages by promoting fusion between the parasitophorous vacuole and mature host lysosomes upon internalization. Since trypomastigotes can evade the lytic pathway, the earliest steps of endocytosis, such as early endosome fusion, may be affected. To test this hypothesis, we used an in vitro early endosome fusion assay. Our results show that trypomastigote-infected macrophage cytosols cannot promote fusion between early endosomes, compared to mock-infected cytosols (heat-killed trypomastigotes were used in the parasite-macrophage interaction assay). GTP gamma S addition potentiates the fusogenic activity driven by trypomastigote-infected macrophage cytosol-mediated assays, unlike the biphasic fusogenic effect obtained with GTP gamma S treatment of macrophage cytosol controls. Calcium-stimulated early endosome fusogenic processes are not affected in the assays mediated by infected macrophage cytosol. We conclude that GTP-regulated factors, and not calcium-regulated elements, are involved in the inhibition of the early endosome fusogenic process by the trypomastigote-infected macrophage cytosol. This primary impediment to the progress of a normal endocytosis may be a relevant step required for the lysosomal recruitment-fusion of the host lysosomes upon trypomastigote infection and further survival of the parasite within its host.
Ochatt
C M
CM
Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI), Buenos Aires, Argentina. ochattc@rockvax.rockefeller.edu
Mayorga
L S
LS
Isola
E L
EL
Wilkowsky
S
S
Torres
H N
HN
Téllez-Iñón
M T
MT
D000818:Animals; D002118:Calcium / Q000502:physiology; D003600:Cytosol / Q000469:parasitology / Q000502:physiology*; D011992:Endosomes / Q000502:physiology*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000502:physiology; D008264:Macrophages / Q000469:parasitology* / Q000648:ultrastructure; D008561:Membrane Fusion / Q000502:physiology*; D051379:Mice; D014349:Trypanosoma cruzi / Q000502:physiology*
D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D002118:Calcium