Ubiquitination and degradation of homeodomain-interacting protein kinase 2 by WD40 repeat/SOCS box protein WSB-1

J Biol Chem. 2008 Feb 22;283(8):4682-9. doi: 10.1074/jbc.M708873200. Epub 2007 Dec 19.


Homeodomain-interacting protein kinase 2 (HIPK2) is a member of the nuclear protein kinase family, which induces both p53- and CtBP-mediated apoptosis. Levels of HIPK2 were increased by UV irradiation and cisplatin treatment, thereby implying the degradation of HIPK2 in cells under normal conditions. Here, we indicate that HIPK2 is ubiquitinated and degraded by the WD40-repeat/SOCS box protein WSB-1, a process that is blocked under DNA damage conditions. Yeast two-hybrid screening was conducted to identify the proteins that interact with HIPK2. WSB-1, an E3 ubiquitin ligase, was characterized as an HIPK2-interacting protein. The coexpression of WSB-1 resulted in the degradation of HIPK2 via its C-terminal region. Domain analysis of WSB-1 showed that WD40-repeats and the SOCS box were required for its interaction with and degradation of HIPK2, respectively. In support of the degradation of HIPK2 by WSB-1, HIPK2 was polyubiquitinated by WSB-1 in vitro and in vivo. The knockdown of endogenous WSB-1 with the expression of short hairpin RNA against WSB-1 increases the stability of endogenous HIPK2 and resulted in the accumulation of HIPK2. The ubiquitination and degradation of HIPK2 by WSB-1 was inhibited completely via the administration of DNA damage reagents, including Adriamycin and cisplatin. These findings effectively illustrate the regulatory mechanisms by which HIPK2 is maintained at a low level, by WSB-1 in cells under normal conditions, and stabilized by genotoxic stresses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Oxidoreductases / genetics
  • Alcohol Oxidoreductases / metabolism
  • Antibiotics, Antineoplastic / pharmacology
  • Apoptosis / drug effects
  • Apoptosis / physiology
  • Apoptosis / radiation effects
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Cell Line
  • Cisplatin / pharmacology
  • DNA Damage / drug effects
  • DNA Damage / physiology
  • DNA Damage / radiation effects
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Doxorubicin / pharmacology
  • Humans
  • Protein Processing, Post-Translational / drug effects
  • Protein Processing, Post-Translational / physiology*
  • Protein Processing, Post-Translational / radiation effects
  • Protein Structure, Tertiary / physiology
  • Protein-Serine-Threonine Kinases / genetics
  • Protein-Serine-Threonine Kinases / metabolism*
  • Proteins / genetics
  • Proteins / metabolism*
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Protein p53 / metabolism
  • Ubiquitination / drug effects
  • Ubiquitination / physiology*
  • Ubiquitination / radiation effects
  • Ultraviolet Rays


  • Antibiotics, Antineoplastic
  • Carrier Proteins
  • DNA-Binding Proteins
  • Proteins
  • Tumor Suppressor Protein p53
  • WSB1 protein, human
  • Doxorubicin
  • Alcohol Oxidoreductases
  • C-terminal binding protein
  • HIPK2 protein, human
  • Protein-Serine-Threonine Kinases
  • Cisplatin