Cell adhesion and Ca2+ signaling activity in stably transfected Trypanosoma cruzi epimastigotes expressing the metacyclic stage-specific surface molecule gp82.
Metacyclic trypomastigotes of Trypanosoma cruzi express a developmentally regulated 82-kDa surface glycoprotein (gp82) that has been implicated in host cell invasion. gp82-mediated interaction of metacyclic forms with target cells induces in both cells activation of the signal transduction pathways, leading to intracellular Ca(2+) mobilization, which is required for parasite internalization. Noninfective epimastigotes do not express detectable levels of gp82 and are unable to induce a Ca(2+) response. We stably transfected epimastigotes with a T. cruzi expression vector carrying the metacyclic stage gp82 cDNA. These transfectants produced a functional gp82, which bound to and triggered a Ca(2+) response in HeLa cells, in the same manner as the metacyclic trypomastigote gp82. Such properties were not found in epimastigotes transfected with the plasmid vector alone. Epimastigotes expressing gp82 on the surface adhered to HeLa cells but were not internalized. Treatment of gp82-expressing epimastigotes with forskolin, an activator of adenylyl cyclase that increases the metacyclic trypomastigote entry into target cells, did not promote parasite internalization. P175, an intracellular tyrosine phosphorylated protein, which appears to play a role in gp82-dependent signaling cascade in metacyclic forms, was undetectable in epimastigotes, either transfected or not with pTEX-gp82. Overall, our results indicate that gp82 is required but not sufficient for target cell invasion.
Manque
Patricio M
PM
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de Sao Paulo, Sao Paulo, Brazil.
Neira
Ivan
I
Atayde
Vanessa D
VD
Cordero
Esteban
E
Ferreira
Alice T
AT
da Silveira
José Franco
JF
Ramirez
Marcel
M
Yoshida
Nobuko
N
D000268:Adhesiveness; D000818:Animals; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling; D006367:HeLa Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000502:physiology*; D051379:Mice; D009893:Opossums; D010766:Phosphorylation; D015800:Protozoan Proteins / Q000502:physiology*; D014162:Transfection; D014349:Trypanosoma cruzi / Q000472:pathogenicity*
D008562:Membrane Glycoproteins; D015800:Protozoan Proteins; D002118:Calcium