Regulation of Trypanosoma cruzi invasion of nonphagocytic cells by the endocytically active GTPases dynamin, Rab5, and Rab7.
During invasion of nonphagocytic cells by Trypanosoma cruzi (T. cruzi), host cell lysosomes are recruited to the plasma membrane attachment site followed by lysosomal enzyme secretion. The membrane trafficking events involved in invasion have not been delineated. We demonstrate here that T. cruzi invasion of nonphagocytic cells was completely abolished by overexpression of a dominant negative mutant of dynamin. Likewise, overexpression of a dominant negative mutant of Rab5, the rate-limiting GTPase for endocytosis, resulted in reduced infection rates compared with cells expressing Rab5 wild-type. Moreover, cells expressing the activated mutant of Rab5 experienced higher infection rates. A similar pattern was also observed when Rab7-transfected cells were examined. Confocal microscopy experiments showed that parasites colocalized with green fluorescent protein-Rab5-positive early endosomes after 5 min of invasion. These data clearly indicate that newly forming T. cruzi phagosomes first interact with an early endosomal compartment and subsequently with other late component markers before lysosomal interaction occurs.
Wilkowsky
S E
SE
Departamento de Microbiología, Universidad de Buenos Aires, Buenos Aires, 1121, Argentina.
Barbieri
M A
MA
Stahl
P D
PD
Isola
E L D
EL
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D034281:Dynamins; D004705:Endocytosis; D011992:Endosomes / Q000201:enzymology*; D020558:GTP Phosphohydrolases / Q000502:physiology*; D006367:HeLa Cells; D006790:Host-Parasite Interactions; D006801:Humans; D008247:Lysosomes / Q000737:chemistry; D018613:Microscopy, Confocal; D008954:Models, Biological; D010586:Phagocytes / Q000502:physiology; D014349:Trypanosoma cruzi / Q000472:pathogenicity*; D014617:Vacuoles / Q000201:enzymology / Q000469:parasitology; D020691:rab GTP-Binding Proteins / Q000502:physiology*; D020696:rab5 GTP-Binding Proteins / Q000502:physiology*; D000090802:rab7 GTP-Binding Proteins
D000090802:rab7 GTP-Binding Proteins; C000716463:rab7 GTP-binding proteins, human; D020558:GTP Phosphohydrolases; D020691:rab GTP-Binding Proteins; D020696:rab5 GTP-Binding Proteins; D034281:Dynamins