p53 Stabilization and Accumulation Induced by Human Vaccinia-Related Kinase 1

Abstract

Variations in intracellular levels of p53 regulate many cellular functions and determine tumor susceptibility. Major mechanisms modulating p53 levels include phosphorylation and interaction of p53 with specific ubiquitin ligases that promote its degradation. N-terminal phosphorylation regulates the interaction of p53 with several regulatory molecules. Vaccinia-related kinase 1 (VRK1) is the prototype of a new Ser-Thr kinase family in the human kinome. VRK1 is located in the nucleus outside the nucleolus. Overexpression of VRK1 increases the stability of p53 by a posttranslational mechanism leading to its accumulation by a mechanism independent of the Chk2 kinase. Catalytically inactive VRK1 protein (a K179E mutant) does not induce p53 accumulation. VRK1 phosphorylates human p53 in Thr18 and disrupts p53-Mdm2 interaction in vitro, although a significant decrease in p53 ubiquitination by Mdm2 in vivo was not detected. VRK1 kinase does not phosphorylate Mdm2. VRK1-mediated p53 stabilization was also detected in Mdm2^−/− cells. VRK1 also has an additive effect with MdmX or p300 to stabilize p53, and p300 coactivation and acetylation of p53 is enhanced by VRK1. The p53 stabilized by VRK1 is transcriptionally active. Suppression of VRK1 expression by specific small interfering RNA provokes several defects in proliferation, situating the protein in the regulation of this process. VRK1 might function as a switch controlling the proteins that interact with p53 and thus modifying its stability and activity. We propose VRK1 as the first step in a new pathway regulating p53 activity during cell proliferation.

We thank D. Lane, S. Lain, T. Hupp, D. Meek, K. J. Pietenpol, G. Lozano, J. Bartek, S. Berberich, M. Scheffner, A. Zantema, T. Halozanetis, and M. Oren for providing us with reagents used in this work.

F.M.V. and A. S. were supported by fellowships from Fundación Ramón Areces and the I3P program of the Consejo Superior de Investigaciones Científicas. This work was funded by grants from the Ministerio de Educación y Ciencia (SAF2000-0169 and SAF2004-2900), the Fondo de Investigación Sanitaria (FIS-PI02-0585), the Junta de Castilla y León (CSI18-03), the Fundación de Investigación Médica MMA, and the Fundación Memoria Samuel Solórzano Barruso.