Up-regulation of actin mRNA and reorganization of the cytoskeleton in Entamoeba histolytica trophozoites

Actin mRNA levels were measured in Entamoeba histolytica trophozoites after experimentally inducing changes in the organization of the cytoskeleton. The treatment of trophozoites with forskolin, N-6,2'-O-dibutyryl-adenosine 3':5'-cyclic monophosphate, and phorbol myristate acetate induced the organization of actin into multiple dots and defined structures with a concomitant increase in F-actin content. Cytochalasin D elicited polarization of the structured actin and formation of aggregates, as well as an increment in F-actin. Simultaneously, up-regulation of actin mRNA levels was produced by all the drugs. De novo synthesis of actin mRNA, as measured by nuclear run-ons, showed increased transcription of actin mRNA. On the other hand, treatment of cells with actinomycin D blocked the elevation of actin mRNA synthesis induced by forskolin, dibutyryl cyclic AMP, or cytochalasin D whereas, the increment induced by PMA was not affected. These data indicate a regulatory control of actin mRNA synthesis at the transcriptional level by forskolin, dibutyryl cyclic AMP and cytochalasin D, and transcriptional as well as post-transcriptional controls by phorbol myristate acetate. The experiments presented here suggest the possibility that, regulation of actin mRNA transcription in E. histolytica trophozoites is linked to growth conditions, that are accompanied by reorganization of the actin cytoskeleton and thus, related to the motility and invasiveness of the parasite.