Activation-loop autophosphorylation is mediated by a novel transitional intermediate form of DYRKs

Cell. 2005 Jun 17;121(6):925-36. doi: 10.1016/j.cell.2005.03.034.


Autophosphorylation of a critical residue in the activation loop of several protein kinases is an essential maturation event required for full enzyme activity. However, the molecular mechanism by which this happens is unknown. We addressed this question for two dual-specificity tyrosine-phosphorylation-regulated protein kinases (DYRKs), as they autophosphorylate their activation loop on an essential tyrosine but phosphorylate their substrates on serine and threonine. Here we demonstrate that autophosphorylation of the critical activation-loop tyrosine is intramolecular and mediated by the nascent kinase passing through a transitory intermediate form. This DYRK intermediate differs in residue and substrate specificity, as well as sensitivity to small-molecule inhibitors, compared with its mature counterpart. The intermediate's characteristics are lost upon completion of translation, making the critical tyrosine autophosphorylation a "one-off" inceptive event. This mechanism is likely to be shared with other kinases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism*
  • Dyrk Kinases
  • Insecta
  • Phosphorylation
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism*
  • Protein Structure, Tertiary / genetics
  • Protein Structure, Tertiary / physiology
  • Protein-Tyrosine Kinases / genetics
  • Protein-Tyrosine Kinases / metabolism*
  • Serine / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Threonine / metabolism
  • Tyrosine / metabolism


  • Drosophila Proteins
  • Threonine
  • Tyrosine
  • Serine
  • mnb protein, Drosophila
  • Protein-Tyrosine Kinases
  • Protein Serine-Threonine Kinases
  • DYRK2 protein, Drosophila