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Abstract:

The ASR (for ABA/water stress/ripening) protein family, first described in tomato as nuclear and involved in adaptation to dry climates, is widespread in the plant kingdom, including crops of high agronomic relevance. We show both nuclear and cytosolic localization for ASR1 (the most studied member of the family) in histological plant samples by immunodetection, typically found in small proteins readily diffusing through nuclear pores. Indeed, a nuclear localization was expected based on sorting prediction software, which also highlight a monopartite nuclear localization signal (NLS) in the primary sequence. However, here we prove that such an "NLS" of ASR1 from tomato is dispensable and non-functional, being the transport of the protein to the nucleus due to simple diffusion across nuclear pores. We attribute such a targeting deficiency to the misplacing in that cryptic NLS of two conserved contiguous lysine residues. Based on previous in vitro experiments regarding quaternary structure, we also carried out live cell imaging assays through confocal microscopy to explore dimer formation in planta. We found homodimers in both the cytosol and the nucleus and demonstrated that assembly of both subunits together can occur in the cytosol, giving rise to translocation of preformed dimers. The presence of dimers was further corroborated by means of in vivo crosslinking of nuclei followed by SDS-PAGE. © 2012 Ricardi et al.

Registro:

Documento: Artículo
Título:Nuclear import and dimerization of tomato ASR1, a water stress-inducible protein exclusive to plants
Autor:Ricardi, M.M.; Guaimas, F.F.; González, R.M.; Burrieza, H.P.; López-Fernández, M.P.; Jares-Erijman, E.A.; Estévez, J.M.; Iusem, N.D.
Filiación:Instituto de Fisiología, Biología Molecular y Neurociencias (IFIByNE-CONICET), Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
Departamento de Química Orgánica and CIHIDECAR, CONICET, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
Departamento de Biodiversidad y Biología Experimental, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
Laboratorio de Fisiología y Biología Molecular (LFBM), Departamento de Fisiología, Biología Molecular y Celular (FBMC), Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
Palabras clave:chaperone; dimer; homodimer; lysine; protein ASR1; unclassified drug; article; cell assay; cell nucleus; confocal microscopy; controlled study; cytosol; dimerization; immunodetection; in vitro study; in vivo study; nonhuman; nuclear import; nuclear localization signal; nuclear pore; polyacrylamide gel electrophoresis; protein assembly; protein cross linking; protein localization; protein quaternary structure; protein transport; tomato; water stress; Active Transport, Cell Nucleus; Cell Nucleus; Cytosol; Dehydration; Lycopersicon esculentum; Nuclear Localization Signals; Plant Proteins; Protein Multimerization
Año:2012
Volumen:7
Número:8
DOI: http://dx.doi.org/10.1371/journal.pone.0041008
Título revista:PLoS ONE
Título revista abreviado:PLoS ONE
ISSN:19326203
CAS:lysine, 56-87-1, 6899-06-5, 70-54-2; Nuclear Localization Signals; Plant Proteins
PDF:https://bibliotecadigital.exactas.uba.ar/download/paper/paper_19326203_v7_n8_p_Ricardi.pdf
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19326203_v7_n8_p_Ricardi

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Citas:

---------- APA ----------
Ricardi, M.M., Guaimas, F.F., González, R.M., Burrieza, H.P., López-Fernández, M.P., Jares-Erijman, E.A., Estévez, J.M.,..., Iusem, N.D. (2012) . Nuclear import and dimerization of tomato ASR1, a water stress-inducible protein exclusive to plants. PLoS ONE, 7(8).
http://dx.doi.org/10.1371/journal.pone.0041008
---------- CHICAGO ----------
Ricardi, M.M., Guaimas, F.F., González, R.M., Burrieza, H.P., López-Fernández, M.P., Jares-Erijman, E.A., et al. "Nuclear import and dimerization of tomato ASR1, a water stress-inducible protein exclusive to plants" . PLoS ONE 7, no. 8 (2012).
http://dx.doi.org/10.1371/journal.pone.0041008
---------- MLA ----------
Ricardi, M.M., Guaimas, F.F., González, R.M., Burrieza, H.P., López-Fernández, M.P., Jares-Erijman, E.A., et al. "Nuclear import and dimerization of tomato ASR1, a water stress-inducible protein exclusive to plants" . PLoS ONE, vol. 7, no. 8, 2012.
http://dx.doi.org/10.1371/journal.pone.0041008
---------- VANCOUVER ----------
Ricardi, M.M., Guaimas, F.F., González, R.M., Burrieza, H.P., López-Fernández, M.P., Jares-Erijman, E.A., et al. Nuclear import and dimerization of tomato ASR1, a water stress-inducible protein exclusive to plants. PLoS ONE. 2012;7(8).
http://dx.doi.org/10.1371/journal.pone.0041008