The R2R3 Myb protein family in Entamoeba histolytica.

The MYB DNA-binding domain is conserved in vertebrates, plants, and fungi. This domain mediates the DNA-binding activity of proteins (that have transcription factor activity) in a sequence-specific manner and is also used for the protection of telomeric regions. The MYB DNA-binding domain contains three imperfect conserved repeats of 52 amino acids (R1, R2, and R3). Within each repeat, there are three tryptophans that are separated by 18 or 19 amino acids. In order to understand the role of Myb transcription factors in Entamoeba histolytica, we searched for MYB DNA-binding domain containing proteins using the amino acid sequence of human c-Myb as the query. We found 34 putative MYB DNA-binding domain containing proteins, which clustered into three monophyletic groups. Family I members conserve only the R2 and R3 repeats in their MYB DNA-binding domain and were dubbed in this report as EhMybR2R3. Family II includes single-repeat proteins related to human telomeric binding proteins. Family III is predicted to comprise proteins with one single repeat where the region corresponding to the conserved tryptophan of the third alpha helix is replaced by a (S)/(T)HAQK(Y)/(F)F motif; this family was named EhMybSHAQKYF. In this work, we focused on proteins that belong to the EhMybR2R3 family. RT-PCR analysis showed that EhMybR2R3 genes were differentially expressed in trophozoites grown in basal culture conditions. Purified rEhMyb10 protein, belonging to the EhMybR2R3 family, was able to bind a consensus Myb recognition element in vitro. In addition, using nuclear extracts from trophozoites of E. histolytica, we were able to detect Myb DNA-binding activity to this sequence. Our in silico surveys demonstrated that this consensus sequence is present in E. histolytica gene promoters. Interestingly, these promoters include different families of genes that are related to signal transduction, vesicular transport, heat shock response, and virulence. Thus, Myb putative transcription factors in E. histolytica could be involved in the transcriptional regulation of genes participating in several different pathways.