Characterization of an Iron-responsive Promoter in the Protozoan Pathogen Trichomonas vaginalis

Iron has been shown to regulate transcription in the protozoan pathogen Trichomonas vaginalis. In this study, a DNA transfection system was developed to monitor ap65-1 promoter activity in response to changing iron supply. In conjunction with electrophoretic mobility shift assay, iron-induced transcription of the ap65-1 gene was shown to be regulated by multiple closely spaced DNA elements spanning an iron-responsive region (-110/-54), including an iron-responsive DNA element ((-98)AGATAACGA(-90)), which overlaps with a 3'-MYB-like protein binding sequence ((-95)TAACGATAT(-87)), and three nearby T-rich sequences ((-110)ATTTTT(-105), (-78)ATTATT(-73), and (-59)ATTTTT(-54)). 5'- and 3'-flanking sequences of the iron-responsive region were shown to regulate basal transcription. A distal DNA regulatory region was shown to enhance both basal and iron-induced transcription. These findings delineate the DNA regulatory elements and nuclear proteins involving in iron-induced transcription of the ap65-1 gene, which provide useful tools for the future study of transcriptional regulation in T. vaginalis.