RESUMO
Cytoskeleton remodeling can be regulated, among other mechanisms, by lysine acetylation. The role of acetylation on cytoskeletal and other proteins of Entamoeba histolytica has been poorly studied. Dynamic rearrangements of the actin cytoskeleton are crucial for amebic motility and capping formation, processes that may be effective means of evading the host immune response. Here we report the possible effect of acetylation on the actin cytoskeleton dynamics and in vivo virulence of E. histolytica. Using western blot, immunoprecipitation, microscopy assays, and in silico analysis, we show results that strongly suggest that the increase in Aspirin-induced cytoplasm proteins acetylation reduced cell movement and capping formation, likely as a consequence of alterations in the structuration of the actin cytoskeleton. Additionally, intrahepatic inoculation of Aspirin-treated trophozoites in hamsters resulted in severe impairment of the amebic virulence. Taken together, these results suggest an important role for lysine acetylation in amebic invasiveness and virulence.
Assuntos
Citoesqueleto de Actina/metabolismo , Entamoeba histolytica/metabolismo , Entamoeba histolytica/patogenicidade , Lisina/metabolismo , Acetilação/efeitos dos fármacos , Citoesqueleto de Actina/efeitos dos fármacos , Actinas/metabolismo , Sequência de Aminoácidos , Animais , Aspirina/farmacologia , Sítios de Ligação , Cricetinae , Citocalasina D/farmacologia , Entamoeba histolytica/crescimento & desenvolvimento , Entamoeba histolytica/ultraestrutura , Masculino , Simulação de Acoplamento Molecular , Dados de Sequência Molecular , Movimento/efeitos dos fármacos , Parasitos/efeitos dos fármacos , Parasitos/crescimento & desenvolvimento , Polimerização/efeitos dos fármacos , Trofozoítos/efeitos dos fármacos , Trofozoítos/crescimento & desenvolvimento , Trofozoítos/ultraestrutura , VirulênciaRESUMO
Protein kinases (PKs) of parasitic protozoa are being evaluated as drug targets. A large number of protein kinases within the protein kinome of Entamoeba histolytica strongly suggest that protein phosphorylation is a key component of pathogenesis regulation by this parasite. PI3 K and Src are kinases previously described in this parasite, but their role is poorly understood. Here, the effect of Src-1-inhibitor and PI3 K inhibitor (Wortmannin) on the virulence factors of E. histolytica was evaluated. Results show that both inhibitors affect the actin cytoskeleton and the amoebic movement. Also, the proteolytic activity is diminished by Wortmannin, but not by Src-inhibitor-1; however, the phagocytic capacity is diminished by Wortmannin and Src-1-inhibitor. Finally, we found that the virulence in vivo of E. histolytica is affected by Wortmannin but not by Src-1-inhibitor. This study opens the way for the design of anti-amoebic drugs based on kinase inhibition.