p53 is phosphorylated in vitro and in vivo by the delta and epsilon isoforms of casein kinase 1 and enhances the level of casein kinase 1 delta in response to topoisomerase-directed drugs

U. Knippschild, D. M. Milne, L. E. Campbell, A. J. DeMaggio, E. Christenson, M. F. Hoekstra, D. W. Meek

    Research output: Contribution to journalArticlepeer-review

    137 Citations (Scopus)

    Abstract

    The p53 tumour suppressor protein plays a key role in the integration of stress signals. Multi-site phosphorylation of p53 may play an integral part in the transmission of these signals and is catalysed by many different protein kinases including an unidentified p53-N-terminus-targeted protein kinase (p53NK) which phosphorylates a group of sites at the N-terminus of the protein. In this paper, we present evidence that the delta and epsilon isoforms of casein kinase 1 (CK1d and CK1e) show identical features to p53NK and can phosphorylate p53 both in vitro and in vivo. Recombinant, purified glutathione S-transferase (GST)-CK1d and GST-CK1e fusion proteins each phosphorylate p53 in vitro at serines 4, 6 and 9, the sites recognised by p53NK. Furthermore, p53NK (i) co-purifies with CK1d/e, (ii) shares identical kinetic properties to CK1delta/epsilon, and (iii) is inhibited by a CK1d/e-specific inhibitor (IC261). In addition, CK1d is also present in purified preparations of p53NK as judged by immunoanalysis using a CK1d-specific monoclonal antibody. Treatment of murine SV3T3 cells with IC261 specifically blocked phosphorylation in vivo of the CK1d/e phosphorylation sites in p53, indicating that p53 interacts physiologically with CK1d and/or CK1e. Similarly, over-expression of a green fluorescent protein (GFP)-CK1d fusion protein led to hyper-phosphorylation of p53 at its N-terminus. Treatment of MethAp53ts cells with the topoisomerase-directed drugs etoposide or camptothecin led to increases in both CK1d-mRNA and -protein levels in a manner dependent on the integrity of p53. These data suggest that p53 is phosphorylated by CK1d and CK1e and additionally that there may be a regulatory feedback loop involving p53 and CK1d.

    Original languageEnglish
    Pages (from-to)1727-1736
    Number of pages10
    JournalOncogene
    Volume15
    Issue number14
    Publication statusPublished - 2 Oct 1997

    Fingerprint Dive into the research topics of 'p53 is phosphorylated <em>in vitro</em> and <em>in vivo</em> by the delta and epsilon isoforms of casein kinase 1 and enhances the level of casein kinase 1 delta in response to topoisomerase-directed drugs'. Together they form a unique fingerprint.

    Cite this