Biochemical quantitation of the eIF5A hypusination in Arabidopsis thaliana uncovers ABA-dependent regulation
Belda-Palazón, Borja (Instituto de Biología Molecular y Celular de Plantas)
Nohales, María A. (Instituto de Biología Molecular y Celular de Plantas)
Rambla, José L. (Instituto de Biología Molecular y Celular de Plantas)
Aceña, José L. (Centro de Investigación Príncipe Felipe (València))
Delgado, Oscar (Centro de Investigación Príncipe Felipe (València))
Fustero, Santos (Centro de Investigación Príncipe Felipe (València))
Martínez, M. Carmen (María Carmen) (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Granell, Antonio (Instituto de Biología Molecular y Celular de Plantas)
Carbonell, Juan (Instituto de Biología Molecular y Celular de Plantas)
Ferrando, Alejandro (Instituto de Biología Molecular y Celular de Plantas)

Fecha:	2014
Resumen:	The eukaryotic translation elongation factor eIF5A is the only protein known to contain the unusual amino acid hypusine which is essential for its biological activity. This post-translational modification is achieved by the sequential action of the enzymes deoxyhypusine synthase (DHS) and deoxyhypusine hydroxylase (DOHH). The crucial molecular function of eIF5A during translation has been recently elucidated in yeast and it is expected to be fully conserved in every eukaryotic cell, however the functional description of this pathway in plants is still sparse. The genetic approaches with transgenic plants for either eIF5A overexpression or antisense have revealed some activities related to the control of cell death processes but the molecular details remain to be characterized. One important aspect of fully understanding this pathway is the biochemical description of the hypusine modification system. Here we have used recombinant eIF5A proteins either modified by hypusination or non-modified to establish a bi-dimensional electrophoresis (2D-E) profile for the three eIF5A protein isoforms and their hypusinated or unmodified proteoforms present in Arabidopsis thaliana. The combined use of the recombinant 2D-E profile together with 2D-E/western blot analysis from whole plant extracts has provided a quantitative approach to measure the hypusination status of eIF5A. We have used this information to demonstrate that treatment with the hormone abscisic acid produces an alteration of the hypusine modification system in Arabidopsis thaliana. Overall this study presents the first biochemical description of the post-translational modification of eIF5A by hypusination which will be functionally relevant for future studies related to the characterization of this pathway in Arabidopsis thaliana.
Ayudas:	Ministerio de Ciencia e Innovación MINECO/BIO2009-11818 Ministerio de Ciencia e Innovación MINECO/BIO2011-23828
Nota:	Altres ajuts: GV/ACIF/2010/085
Derechos:	Tots els drets reservats.
Lengua:	Català
Documento:	Article ; recerca ; Versió publicada
Materia:	Spermidine ; Hypusine ; EIF5A ; 2D-electrophoresis ; Abscisic acid
Publicado en:	Frontiers in plant science, Vol. 5 (2014) , ISSN 1664-462X

DOI: 10.3389/fpls.2014.00202
PMID: 24904603

11 p, 2.3 MB

El registro aparece en las colecciones:
Artículos > Artículos de investigación
Artículos > Artículos publicados