L10 ribosomal protein from Entamoeba histolytica share structural and functional homologies with QM/Jif-1: proteins with extraribosomal functions.

Abstract

In the present work, the complete amino acid sequence of the Entamoeba histolytica ribosomal protein L10 (EhL10) is reported. cDNA of 630bp revealed an open reading frame that encodes a protein of 210 amino acids. Analysis of EhL10 ribosomal protein revealed 75% similarity and 57% identity with QM protein from Homo sapiens and 78 and 60%, respectively, with Arabidopsis thaliana. Western blot analysis of ribosomal proteins from E. histolytica showed that EhL10 protein is part of the ribosomal complex. Immunofluorescence analysis of EhL10 distribution in a transfected E. histolytica strain showed that EhL10 protein was mainly localized in the nucleus of trophozoites. Overexpression of EhL10 ribosomal protein in trophozoites transfected with the pExEhNeo/EhL10 vector exhibited a 60% reduction in cellular growth. DNA mobility-shift assays demonstrated that EhL10 ribosomal protein was able to destabilize the activating protein 1 (AP-1) complex binding specifically to the c-Jun-like protein. It is proposed in this study that the complex formation of EhL10 with c-Jun-like protein interferes with transcriptional activation of genes controlled by Jun (i.e. gene involved in cell growth). It is also being reported identification of a member of the AP-1 complex, the c-Jun-like protein, in nuclear extracts of E. histolytica using human-specific antibodies against this protein. The observations suggest that EhL10 may have an extraribosomal function in E. histolytica involved in suppression of cell proliferation in E. histolytica similar to the QM protein.