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Nucleic Acids Research 2005 33(5):1722-1736; doi:10.1093/nar/gki314
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Published online 22 March 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Article

Transcriptional regulation of two stage-specifically expressed genes in the protozoan parasite Toxoplasma gondii

Michael K. Kibe, Alexandra Coppin, Najoua Dendouga, Gabrielle Oria, Edwige Meurice, Marlène Mortuaire, Edwige Madec and Stanislas Tomavo*

Equipe de Parasitologie Moléculaire UGSF-IFR 118, CNRS UMR 8576 Université des Sciences et Technologies de Lille 59650 Villeneuve d'Ascq, France

*To whom correspondence should be addressed at Equipe de Parasitologie Moléculaire, Laboratoire de Chimie Biologique, CNRS UMR 8576, Bâtiment C9, Université des Sciences et Technologies de Lille, 59650 Villeneuve d'Ascq, France. Tel: +33 03 20 43 69 41; Fax: +33 03 20 65 55; Email: Stan.Tomavo{at}univ-lille1.fr

Received February 9, 2005. Revised March 2, 2005. Accepted March 2, 2005.

The protozoan parasite Toxoplasma gondii differentially expresses two distinct enolase isoenzymes known as ENO1 and ENO2, respectively. To understand differential gene expression during tachyzoite to bradyzoite conversion, we have characterized the two T.gondii enolase promoters. No homology could be found between these sequences and no TATA or CCAAT boxes were evident. The differential activation of the ENO1 and ENO2 promoters during tachyzoite to bradyzoite differentiation was investigated by deletion analysis of 5'-flanking regions fused to the chloramphenicol acetyltransferase reporter followed by transient transfection. Our data indicate that in proliferating tachyzoites, the repression of ENO1 involves a negative distal regulatory region (nucleotides –1245 to –625) in the promoter whereas a proximal regulatory region in the ENO2 promoter directs expression at a low level. In contrast, the promoter activity of ENO1 is highly induced following the conversion of tachyzoites into resting bradyzoites. The ENO2 promoter analysis in bradyzoites showed that there are two upstream repression sites (nucleotides –1929 to –1067 and –456 to –222). Furthermore, electrophoresis mobility shift assays demonstrated the presence of DNA-binding proteins in tachyzoite and bradyzoite nuclear lysates that bound to stress response elements (STRE), heat shock-like elements (HSE) and other cis-regulatory elements in the upstream regulatory regions of ENO1 and ENO2. Mutation of the consensus AGGGG sequence, completely abolished protein binding to an oligonucleotide containing this element. This study defines the first characterization of cis-regulatory elements and putative transcription factors involved in gene regulation of the important pathogen T.gondii.

DDBJ/EMBL/GenBank accession no. AY155668


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