Targets of the Entamoeba histolytica transcription factor URE3-BP

• Published in: PLoS neglected tropical diseases Vol. 2; no. 8; p. e282
• Main Authors: Gilchrist, Carol A, Baba, Duza J, Zhang, Yan, Crasta, Oswald, Evans, Clive, Caler, Elisabet, Sobral, Bruno W S, Bousquet, Christina B, Leo, Megan, Hochreiter, Ameilia, Connell, Sarah K, Mann, Barbara J, Petri, William A
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 27-08-2008; Public Library of Science (PLoS)
• Subjects: 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; Promoter Regions, Genetic; Two-Hybrid System Techniques; Animals; Oligonucleotide Array Sequence Analysis; Regulatory Sequences, Nucleic Acid; Protein Binding; Entamoeba histolytica; Electrophoretic Mobility Shift Assay; Reverse Transcriptase Polymerase Chain Reaction; DNA-Binding Proteins
• ISSN: 1935-2735; 1935-2727; 1935-2735
• DOI: 10.1371/journal.pntd.0000282

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.