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J. Biol. Chem., Vol. 277, Issue 7, 5153-5162, February 15, 2002

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

Chu-Dang Tsai^§, Hsing-Wei Liu^§, and Jung-Hsiang Tai^§¶

From the  Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan 114, Republic of China and the ^§ Division of Infectious Diseases, Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan 11529, Republic of China

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 (⁹⁸AGATAACGA⁹⁰), which overlaps with a 3'-MYB-like protein binding sequence (⁹⁵TAACGATAT⁸⁷), and three nearby T-rich sequences (¹¹⁰ATTTTT¹⁰⁵, ⁷⁸ATTATT⁷³, and ⁵⁹ATTTTT⁵⁴). 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.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AF364546.

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