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The human vaccinia-related kinase 1 (VRK1) phosphorylates threonine-18 within the mdm-2 binding site of the p53 tumour suppressor protein

Oncogene volume 19, pages 3656–3664 (2000)Cite this article

Abstract

The tumour suppressor p53 protein integrates multiple signals regulating cell cycle progression and apoptosis. This regulation is mediated by several kinases that phosphorylate specific residues in the different functional domains of the p53 molecule. The human VRK1 protein is a new kinase related to a poxvirus kinase, and more distantly to the casein kinase 1 family. We have characterized the biochemical properties of human VRK1 from HeLa cells. VRK1 has a strong autophosphorylating activity in several Ser and Thr residues. VRK-1 phosphorylates acidic proteins, such as phosvitin and casein, and basic proteins such as histone 2b and myelin basic protein. Because some transcription factors are regulated by phosphorylation, we tested as substrates the N-transactivation domains of p53 and c-Jun fused to GST. Human c-Jun is not phosphorylated by VRK1. VRK1 phosphorylates murine p53 in threonine 18. This threonine is within the p53 hydrophobic loop (residues 13–23) required for the interaction of p53 with the cleft of its inhibitor mdm-2. The VRK1 C-terminus domain (residues 268–396) that contains a nuclear localization signal targets the protein to the nucleus, as determined by using fusion proteins with the green fluorescent protein. We conclude that VRK1 is an upstream regulator of p53 that belongs to a new signalling pathway.

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Abbreviations

VRK1:

vaccinia related kinase 1

CK1:

casein kinase1

GST:

glutathione-S-transferase

EST:

expressed sequence tag

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Acknowledgements

S Lopez-Borges was supported by a fellowship from Comunidad de Madrid. This work was supported by grants to PA Lazo from Fondo de Investigación Sanitaria (FIS98-0313), Comunidad de Madrid (08.1-0004-98), and Comisión Interministerial de Ciencia y Tecnologia (SAF00/0169).

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Authors and Affiliations

  1. Centro de Investigación del Cáncer, Instituto de Biología Molecular y Celular del Cáncer, Consejo Superior de Investigaciones Científicas, Universidad de Salamanca, Campus Miguel de Unamuno, Salamanca, 37007, Spain

    Susana Lopez-Borges & Pedro A Lazo

  2. Unidad de Genética y Medicina Molecular, CSIC-Centro Nacional de Biología Fundamental, Instituto de Salud Carlos III, Majadahonda, 28220, Spain

    Susana Lopez-Borges & Pedro A Lazo

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  1. Susana Lopez-Borges
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  2. Pedro A Lazo
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Lopez-Borges, S., Lazo, P. The human vaccinia-related kinase 1 (VRK1) phosphorylates threonine-18 within the mdm-2 binding site of the p53 tumour suppressor protein. Oncogene 19, 3656–3664 (2000). https://doi.org/10.1038/sj.onc.1203709

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