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Vazquez, M.P.; Mualem, D.; Bercovich, N.; Stern, M.Z.; Nyambega, B.; Barda, O.; Masiga, D.; Gupta, S.K.; Michaeli, S.; Levin, M.J. "Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1" (2009) Molecular and Biochemical Parasitology. 164(2):137-146
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Abstract:

Early in the assembly of the spliceosome of eukaryotes the branch-point binding protein (BBP, also called SF1) recognizes the branch point sequence, whereas the heterodimer U2AF consisting of a 65 and a 35 kDa subunit, contacts the polypyrimidine tract and the AG splice site, respectively. Herein, we identified, cloned and expressed the Trypanosoma cruzi and Trypanosoma brucei U2AF35, U2AF65 and SF1. Trypanosomatid U2AF65 strongly diverged from yeast and human homologues. On the contrary, trypanosomatid SF1 was conserved but lacked the C-terminal sequence present in the mammalian protein. Yeast two hybrid approaches were used to assess their interactions. The interaction between U2AF35 and U2AF65 was very weak or not detectable. However, as in other eukaryotes, the interaction between U2AF65 and SF1 was strong. At the cellular level, these results were confirmed by fractionation and affinity-selection experiments in which SF1 and U2AF65 co-fractionated in a complex of approximately 400 kDa and U2AF65 was affinity-selected with TAP tagged SF1, but not with TAP tagged U2AF35. Silencing of the three factors affected growth and trans-splicing in the first step of this reaction. Trypanosomes are the first described example of eukaryotic cells in which the interaction of two expressed U2AF factors seemed to be very weak, or even undetectable. © 2009 Elsevier B.V. All rights reserved.

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Documento: Artículo
Título:Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1
Autor:Vazquez, M.P.; Mualem, D.; Bercovich, N.; Stern, M.Z.; Nyambega, B.; Barda, O.; Masiga, D.; Gupta, S.K.; Michaeli, S.; Levin, M.J.
Filiación:Laboratorio de Biología Molecular de la Enfermedad de Chagas, INGEBI- CONICET, Vuelta de Obligado 2490 2P, 1428 Buenos Aires, Argentina
Departamento de Fisiología y Biología Molecular y Celular, Facultad de Ciencias Exactas y Naturales, University of Buenos Aires, Argentina
The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, 52900, Israel
International Center for Insect Physiology and Ecology, P.O.Box 30772-00100, Nairobi, Kenya
Institut Cochin, Departement Maladies Infectieuses, INSERM U567, Paris, F-75014, France
Palabras clave:Trans-splicing; Trypanosoma brucei; Trypanosoma cruzi; U2 auxiliary factor; protein Sf1; protein u2af35; protein U2AF65; protozoal protein; unclassified drug; article; cell fractionation; controlled study; molecular cloning; nonhuman; priority journal; protein analysis; protein expression; protein function; protein localization; protein protein interaction; RNA splicing; Trypanosoma brucei; Trypanosoma cruzi; Amino Acid Sequence; Animals; Cell Fractionation; Cloning, Molecular; Conserved Sequence; Gene Expression; Gene Silencing; Models, Molecular; Molecular Sequence Data; Protein Interaction Mapping; Protein Structure, Tertiary; Protozoan Proteins; RNA Splicing; RNA-Binding Proteins; Sequence Homology, Amino Acid; Trypanosoma brucei brucei; Trypanosoma cruzi; Two-Hybrid System Techniques; Eukaryota; Mammalia; Trypanosoma brucei; Trypanosoma cruzi; Trypanosomatidae
Año:2009
Volumen:164
Número:2
Página de inicio:137
Página de fin:146
DOI: http://dx.doi.org/10.1016/j.molbiopara.2008.12.009
Título revista:Molecular and Biochemical Parasitology
Título revista abreviado:Mol. Biochem. Parasitol.
ISSN:01666851
CODEN:MBIPD
CAS:Protozoan Proteins; RNA-Binding Proteins
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v164_n2_p137_Vazquez

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Citas:

---------- APA ----------
Vazquez, M.P., Mualem, D., Bercovich, N., Stern, M.Z., Nyambega, B., Barda, O., Masiga, D.,..., Levin, M.J. (2009) . Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1. Molecular and Biochemical Parasitology, 164(2), 137-146.
http://dx.doi.org/10.1016/j.molbiopara.2008.12.009
---------- CHICAGO ----------
Vazquez, M.P., Mualem, D., Bercovich, N., Stern, M.Z., Nyambega, B., Barda, O., et al. "Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1" . Molecular and Biochemical Parasitology 164, no. 2 (2009) : 137-146.
http://dx.doi.org/10.1016/j.molbiopara.2008.12.009
---------- MLA ----------
Vazquez, M.P., Mualem, D., Bercovich, N., Stern, M.Z., Nyambega, B., Barda, O., et al. "Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1" . Molecular and Biochemical Parasitology, vol. 164, no. 2, 2009, pp. 137-146.
http://dx.doi.org/10.1016/j.molbiopara.2008.12.009
---------- VANCOUVER ----------
Vazquez, M.P., Mualem, D., Bercovich, N., Stern, M.Z., Nyambega, B., Barda, O., et al. Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1. Mol. Biochem. Parasitol. 2009;164(2):137-146.
http://dx.doi.org/10.1016/j.molbiopara.2008.12.009