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J Biol Chem, Vol. 275, Issue 16, 12051-12060, April 21, 2000

Amastin mRNA Abundance in Trypanosoma cruzi Is Controlled by a 3'-Untranslated Region Position-dependent cis-Element and an Untranslated Region-binding Protein

Bridget C. Coughlin, Santuza M. R. Teixeira^§¶, Louis V. Kirchhoff^§, and John E. Donelson^**

From the Departments of  Biochemistry and ^§ Internal Medicine, University of Iowa and the  Department of Veterans Affairs Medical Center, Iowa City, Iowa 52242

The genome of Trypanosoma cruzi contains tandem arrays of alternating genes encoding amastin and tuzin. Amastin is a surface glycoprotein abundantly expressed on the intracellular mammalian amastigote form of the protozoan parasite, and tuzin is a G-like protein. We demonstrated previously that the amastin-tuzin gene cluster is polycistronically transcribed to an equal extent in all parasite life cycle stages. The steady state level of amastin mRNA, however, is 68-fold more abundant in amastigotes than in epimastigotes. Here we show that the half-life of amastin mRNA is 7 times longer in amastigotes than in epimastigotes. Linker replacement experiments demonstrate that the middle one-third of the 630-nucleotide 3'-untranslated region (UTR) is responsible for the amastin mRNA up-regulation. This positive effect is dependent on the distance of the 3'-UTR segment from the stop codon and the polyadenylation site as well as on its orientation. A protein or protein complex more abundant in amastigotes than in epimastigotes binds to this minimally defined 3'-UTR segment and may be involved in its regulatory function.

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