Journal Article

Targets of the Entamoeba histolytica Transcription Factor URE3-BP

Bousquet, Christina B., Connell, Sarah K., Baba, Duza J., Caler, Elisabet, Mann, Barbara J., Crasta, Oswald, Sobral, Bruno W. S., Hochreiter, Ameilia, Evans, Clive, Zhang, Yan, Leo, Megan, Gilchrist, Carol A., Petri, William A.

2008

Overview

The Entamoeba histolytica transcription factor Upstream Regulatory Element 3-Binding Protein (URE3-BP) is a calcium-responsive regulator of two E. histolytica virulence genes, hgl5 and fdx1. URE3-BP was previously identified by a yeast one-hybrid screen of E. histolytica proteins capable of binding to the sequence TATTCTATT (Upstream Regulatory Element 3 (URE3)) in the promoter regions of hgl5 and fdx1. In this work, precise definition of the consensus URE3 element was performed by electrophoretic mobility shift assays (EMSA) using base-substituted oligonucleotides, and the consensus motif validated using episomal reporter constructs. Transcriptome profiling of a strain induced to produce a dominant-positive URE3-BP was then used to identify additional genes regulated by URE3-BP. Fifty modulated transcripts were identified, and of these the EMSA defined motif T[atg]T[tc][cg]T[at][tgc][tg] was found in over half of the promoters (54% p<0.0001). Fifteen of the URE3-BP regulated genes were potential membrane proteins, suggesting that one function of URE3-BP is to remodel the surface of E. histolytica in response to a calcium signal. Induction of URE3-BP leads to an increase in tranwell migration, suggesting a possible role in the regulation of cellular motility.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Animals / Biochemistry/Transcription and Translation / Bioinformatics / Calcium / DNA-Binding Proteins - genetics / DNA-Binding Proteins - metabolism / DNA-Binding Proteins - physiology / Electrophoretic Mobility Shift Assay / Entamoeba histolytica - genetics / Entamoeba histolytica - metabolism / Experiments / Gene expression / Infectious Diseases/Gastrointestinal Infections / Infectious Diseases/Neglected Tropical Diseases / Infectious Diseases/Protozoal Infections / Infectious Diseases/Tropical and Travel-Associated Diseases / Molecular Biology/Transcription Initiation and Activation / Motility / Mutation / Oligonucleotide Array Sequence Analysis / Promoter Regions, Genetic / Protein Binding / Proteins / Regulatory Sequences, Nucleic Acid - genetics / Reverse Transcriptase Polymerase Chain Reaction / Tropical diseases / Two-Hybrid System Techniques / Yeast / Yeasts