Caractérisation de la protéine kinase PNek1 et de la 14-3-3 P20-5 chez le peuplier

Abstract

Afin d'obtenir une meilleure compréhension des processus de signalisation cellulaire chez le peuplier, quatre isoformes de protéines de type 14-3-3 ont été isolées. La méthode du double hybride a révélé une interaction protéique entre la 14-3-3 P20-5 et PNek1, une protéine kinase de type NIMA. On a alors noté une augmentation de l'accumulation des transcrits de PNek1 lors de l'apparition des cellules mitotiques, soit autour de la transition G2/M. De plus, l'expression constitutive de la fusion GFP-PNek1 chez Arabidopsis a permis de la localiser dans le noyau (incluant le nucléole) et au niveau du cytoplasme. Enfin, chez ces lignées GFP-PNek1 on a remarqué l'apparition d'anomalies morphologiques des fleurs et des siliques ainsi qu'une diminution sévère de la production de graines. En conclusion, ces travaux suggèrent un rôle pour PNek1 dans le cycle cellulaire et ouvrent la voie à la caractérisation des protéines kinases de type NIMA chez les végétaux.

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To obtain a better understanding of cell signaling processes in poplar, four isoforms of 14-3-3 proteins were isolated. The two-hybrid method revealed a protein interaction between 14-3-3 P20-5 and PNek1, a NIMA-like protein kinase. We then noted an increase in the accumulation of PNek1 transcripts during the appearance of mitotic cells, i.e. around the G2/M transition. Furthermore, the constitutive expression of the GFP-PNek1 fusion in Arabidopsis made it possible to localize it in the nucleus (including the nucleolus) and in the cytoplasm. Finally, in these GFP-PNek1 lines we noticed the appearance of morphological anomalies of flowers and siliques as well as a severe reduction in seed production. In conclusion, this work suggests a role for PNek1 in the cell cycle and opens the way to the characterization of NIMA-type protein kinases in plants.