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Abstract:

There are only a few reported nuclear localization signals (NLS) in trypanosomes despite intensive research on nuclear metabolic processes such as mRNA processing and transcription during the recent past. Moreover, there are only two reports for a monopartite (La protein) and bipartite (H2B histone, ESAG8) NLS in Trypanosoma brucei. We decided to investigate a NLS in Trypanosoma cruzi by selecting p14, a small RNA recognition motif (RRM) containing protein involved in the splicing process in the nucleus. Its small size (117 amino acids), and an optimized streamlined workflow for analysis in T. cruzi, allowed us to define a region of basic amino acids (RRKRRR) located at the C-terminus that is necessary for nuclear localization. However, the NLS for p14 appeared to be more complex since the signature RRKRRR alone is necessary but not sufficient to direct heterologous proteins, such as GFP, to the nucleus. Since p14 interacts strongly with splicing factor SF3b155, a much larger protein, we designed a p14 variant unable to interact with it. The results allowed us to discard the notion that p14 is entering the nucleus, or is retained within, as the sole consequence of being part of a larger complex. Extensive mapping showed that all of the information for nuclear import resides within the small p14 protein in a bipartite NLS composed of the signature RRKRRR and a region of the RRM domain. Thus, NLS definition in T. cruzi is more complex than previously described. © 2010 Australian Society for Parasitology Inc.

Registro:

Documento: Artículo
Título:Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi
Autor:Westergaard, G.G.; Bercovich, N.; Reinert, M.D.; Vazquez, M.P.
Filiación:INGEBI-CONICET, Vuelta de Obligado 2490, 2P, Buenos Aires, Argentina
Departamento de Fisiología y Biología Molecular y Celular, Facultad de Ciencias Exactas y Naturales, University of Buenos Aires, Argentina
Max Planck for developmental Biology, Tübingen, Germany
Palabras clave:Gateway vector; NLS; P14; PTREX; Splicing factor; Trypanosoma; cell protein; green fluorescent protein; protein p14; protein SF3b155; unclassified drug; nuclear localization signal; nuclear protein; protozoal protein; recombinant protein; RNA binding protein; amino acid; mapping; protein; RNA; amino acid sequence; article; carboxy terminal sequence; cell nucleus; complex formation; controlled study; genetic transfection; molecular recognition; molecular size; nonhuman; nuclear import; nuclear localization signal; peptide mapping; polymerase chain reaction; protein domain; protein motif; RNA splicing; Trypanosoma cruzi; yeast; enzymology; genetics; metabolism; protein transport; Trypanosoma cruzi; Amino Acid Motifs; Green Fluorescent Proteins; Nuclear Localization Signals; Nuclear Proteins; Protein Transport; Protozoan Proteins; Recombinant Proteins; RNA-Binding Proteins; Trypanosoma cruzi; Trypanosoma; Trypanosoma brucei; Trypanosoma cruzi; Amino Acid Motifs; Green Fluorescent Proteins; Nuclear Localization Signals; Nuclear Proteins; Protein Transport; Protozoan Proteins; Recombinant Proteins; RNA-Binding Proteins; Trypanosoma cruzi
Año:2010
Volumen:40
Número:9
Página de inicio:1029
Página de fin:1035
DOI: http://dx.doi.org/10.1016/j.ijpara.2010.02.011
Título revista:International Journal for Parasitology
Título revista abreviado:Int. J. Parasitol.
ISSN:00207519
CODEN:IJPYB
CAS:Green Fluorescent Proteins, 147336-22-9; Nuclear Localization Signals; Nuclear Proteins; Protozoan Proteins; RNA-Binding Proteins; Recombinant Proteins; Green Fluorescent Proteins; Nuclear Localization Signals; Nuclear Proteins; Protozoan Proteins; Recombinant Proteins; RNA-Binding Proteins
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00207519_v40_n9_p1029_Westergaard

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Citas:

---------- APA ----------
Westergaard, G.G., Bercovich, N., Reinert, M.D. & Vazquez, M.P. (2010) . Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi. International Journal for Parasitology, 40(9), 1029-1035.
http://dx.doi.org/10.1016/j.ijpara.2010.02.011
---------- CHICAGO ----------
Westergaard, G.G., Bercovich, N., Reinert, M.D., Vazquez, M.P. "Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi" . International Journal for Parasitology 40, no. 9 (2010) : 1029-1035.
http://dx.doi.org/10.1016/j.ijpara.2010.02.011
---------- MLA ----------
Westergaard, G.G., Bercovich, N., Reinert, M.D., Vazquez, M.P. "Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi" . International Journal for Parasitology, vol. 40, no. 9, 2010, pp. 1029-1035.
http://dx.doi.org/10.1016/j.ijpara.2010.02.011
---------- VANCOUVER ----------
Westergaard, G.G., Bercovich, N., Reinert, M.D., Vazquez, M.P. Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi. Int. J. Parasitol. 2010;40(9):1029-1035.
http://dx.doi.org/10.1016/j.ijpara.2010.02.011