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Originally published In Press as doi:10.1074/jbc.M209092200 on October 25, 2002

J. Biol. Chem., Vol. 277, Issue 52, 50520-50528, December 27, 2002
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TcUBP-1, an mRNA Destabilizing Factor from Trypanosomes, Homodimerizes and Interacts with Novel AU-rich Element- and Poly(A)-binding Proteins Forming a Ribonucleoprotein Complex*

Iván D'OrsoDagger and Alberto C. C. Frasch§

From the Instituto de Investigaciones Biotecnológicas-Instituto Tecnológico de Chascomús, CONICET-UNSAM, 1650 San Martín, Provincia de Buenos Aires, Argentina

Trypanosomes, protozoan parasites causing worldwide infections in human and animals, mostly regulate protein expression through post-transcriptional mechanisms and not at the transcription initiation level. We have previously identified a Trypanosoma cruzi RNA-binding protein named TcUBP-1. This protein is involved in mRNA destabilization in vivo through binding to AU-rich elements in the 3'-untranslated region of SMUG mucin mRNAs (D'Orso, I., and Frasch, A. C. (2001) J. Biol. Chem. 276, 34801-34809). In this work we show that TcUBP-1 is part of an ~450-kDa ribonucleoprotein complex with a poly(A)-binding protein and a novel 18-kDa RNA-binding protein, named TcUBP-2. Recombinant TcUBP-1 and TcUBP-2 proteins recognize U-rich RNAs with similar specificity and affinity through the ~92-amino acid RNA recognition motif. TcUBPs can homo- and heterodimerize in vitro through the glycine-rich C-terminal region. This interaction was also detected in vivo by co-immunoprecipitation of the ribonucleoprotein complex and using yeast two-hybrid assay. The poly(A)-binding protein identified was shown to disrupt the formation of TcUBP-1, but not TcUBP-2, homodimers in vitro. The possible role of TcUBP-1 ligands in the pathways that govern mRNA-stability and stage-specific expression in trypanosomes is discussed.

* This work was supported in part by grants from the World Bank/UNDP/WHO Special Program for Research and Training in Tropical Diseases (Tropical Disease Research) and by the Agencia Nacional de Promoción Científica y Tecnológica (Argentina).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AF497746.

Dagger Supported in part by an International Research Scholars Grant from the Howard Hughes Medical Institute and a fellowship from the John Simon Guggenheim Memorial Foundation. Researcher from the National Research Council (CONICET), Argentina.

§ Research fellow from the National Research Council (CONICET), Argentina. To whom correspondence should be addressed: IIB-UNSAM, INTI, Av. Gral. Paz s/n, Edificio 24, Casilla de Correo 30, 1650 San Martín, Provincia de Buenos Aires, Argentina. Tel.: 54-11-4580-7255; Fax: 54-11-4752-9639; E-mail: cfrasch@iib.unsam.edu.ar.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.


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