Zipper-interacting protein kinase is involved in regulation of ubiquitination of the androgen receptor, thereby contributing to dynamic transcription complex assembly

Oncogene. 2013 Oct 10;32(41):4981-8. doi: 10.1038/onc.2012.503. Epub 2012 Nov 12.


We have recently identified apoptosis-antagonizing transcription factor (AATF), tumor-susceptibility gene 101 (TSG101) and zipper-interacting protein kinase (ZIPK) as novel coactivators of the androgen receptor (AR). The mechanisms of coactivation remained obscure, however. Here we investigated the interplay and interdependence between these coactivators and the AR using the endogenous prostate specific antigen (PSA) gene as model for AR-target genes. Chromatin immunoprecipitation in combination with siRNA-mediated knockdown revealed that recruitment of AATF and ZIPK to the PSA enhancer was dependent on AR, whereas recruitment of TSG101 was dependent on AATF. Association of AR and its coactivators with the PSA enhancer or promoter occurred in cycles. Dissociation of AR-transcription complexes was due to degradation because inhibition of the proteasome system by MG132 caused accumulation of AR at enhancer/promoter elements. Moreover, inhibition of degradation strongly reduced transcription, indicating that continued and efficient transcription is based on initiation, degradation and reinitiation cycles. Interestingly, knockdown of ZIPK by siRNA had a similar effect as MG132, leading to reduced transcription but enhanced accumulation of AR at androgen-response elements. In addition, knockdown of ZIPK, as well as overexpression of a dominant-negative ZIPK mutant, diminished polyubiquitination of AR. Furthermore, ZIPK cooperated with the E3 ligase Mdm2 in AR-dependent transactivation, assembled into a single complex on chromatin and phosphorylated Mdm2 in vitro. These results suggest that ZIPK has a crucial role in regulation of ubiquitination and degradation of the AR, and hence promoter clearance and efficient transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Androgens / metabolism
  • Apoptosis Regulatory Proteins / metabolism
  • DNA-Binding Proteins / metabolism
  • Endosomal Sorting Complexes Required for Transport / metabolism
  • Gene Knockdown Techniques
  • HEK293 Cells
  • Humans
  • MAP Kinase Kinase Kinases / deficiency
  • MAP Kinase Kinase Kinases / genetics
  • MAP Kinase Kinase Kinases / metabolism*
  • Phosphorylation
  • Proto-Oncogene Proteins c-mdm2 / genetics
  • Proto-Oncogene Proteins c-mdm2 / metabolism
  • RNA, Small Interfering / genetics
  • Receptors, Androgen / deficiency
  • Receptors, Androgen / genetics
  • Receptors, Androgen / metabolism*
  • Regulatory Elements, Transcriptional
  • Repressor Proteins / metabolism
  • Transcription Factors / metabolism
  • Transcription, Genetic*
  • Transcriptional Activation
  • Ubiquitination*


  • AATF protein, human
  • Androgens
  • Apoptosis Regulatory Proteins
  • DNA-Binding Proteins
  • Endosomal Sorting Complexes Required for Transport
  • RNA, Small Interfering
  • Receptors, Androgen
  • Repressor Proteins
  • Transcription Factors
  • Tsg101 protein
  • Proto-Oncogene Proteins c-mdm2
  • MAP Kinase Kinase Kinases
  • mitogen-activated protein kinase kinase kinase 12