Cyclin H binding to the RARalpha activation function (AF)-2 domain directs phosphorylation of the AF-1 domain by cyclin-dependent kinase 7

Proc Natl Acad Sci U S A. 2005 Nov 15;102(46):16608-13. doi: 10.1073/pnas.0505556102. Epub 2005 Nov 7.

Abstract

The transcriptional activity of nuclear retinoic acid receptors (RARs), which act as RAR/retinoid X receptor (RXR) heterodimers, depends on two activation functions, AF-1 and AF-2, which are targets for phosphorylations and synergize for the activation of retinoic acid target genes. The N-terminal AF-1 domain of RARalpha is phosphorylated at S77 by the cyclin-dependent kinase (cdk)-activating kinase (CAK) subcomplex (cdk7/cyclin H/MAT1) of the general transcription factor TFIIH. Here, we show that phosphorylation of S77 governing the transcriptional activity of RARalpha depends on cyclin H binding at a RARalpha region that encompasses loop 8-9 and the N-terminal tip of helix 9 of the AF-2 domain. We propose a model in which the structural constraints of this region control the architecture of the RAR/RXR/TFIIH complex and therefore the efficiency of RARalpha phosphorylation by cdk7. To our knowledge, this study provides the first example of a cooperation between the AF-2 and AF-1 domains of RARs through a kinase complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • Cyclin H
  • Cyclin-Dependent Kinase-Activating Kinase
  • Cyclin-Dependent Kinases / metabolism*
  • Cyclins / metabolism*
  • DNA Primers
  • Models, Molecular
  • Phosphorylation
  • Protein Binding
  • RNA, Small Interfering
  • Receptors, Retinoic Acid / chemistry
  • Receptors, Retinoic Acid / metabolism*
  • Receptors, Retinoic Acid / physiology
  • Retinoic Acid Receptor alpha
  • Reverse Transcriptase Polymerase Chain Reaction
  • Spodoptera
  • Transcription, Genetic / physiology

Substances

  • Cyclin H
  • Cyclins
  • DNA Primers
  • RNA, Small Interfering
  • Receptors, Retinoic Acid
  • Retinoic Acid Receptor alpha
  • Cyclin-Dependent Kinases
  • Cyclin-Dependent Kinase-Activating Kinase