A unique phosphorylation-dependent mechanism for the activation of Ca2+/calmodulin-dependent protein kinase type IV/GR.

Journal Article (Journal Article)

The activity of the Ca2+/calmodulin-dependent protein kinase IV/Gr (CaMKIV/Gr) is shown to be strictly regulated by phosphorylation of three residues both in vitro and in response to antigen receptor-mediated signaling in lymphocytes. One residue, Thr-200, is indispensable for enhancement of Ca2+/calmodulin-dependent basal activity by CaMKIV/Gr kinase. This event requires Ca2+/calmodulin in the full-length CaMKIV/Gr but is Ca2+/calmodulin-independent when a truncated version of CaMKIV/Gr is used as a substrate (DeltaCaMKIV/Gr1-317 (Delta1-317)). The other two residues, Ser12 and Ser13, are apparently autophosphorylated by the Ca2+/calmodulin-bound CaMKIV/Gr. Phosphorylation of neither Ser12-Ser13 nor Thr312 (the residue in a homologous position to Thr286 of CaMKIIalpha influences the development of Ca2+/calmodulin-independent activity or any other property of CaMKIV/Gr examined. Similarly, removal of the NH2-terminal 20 amino acids has no effect on the activation or function of CaMKIV/Gr. However, mutation of both Ser12 and Ser13 residues to Ala in Delta1-317 completely abrogates activity, while individual substitutions have no effect. These results indicate that the NH2-terminal Ser cluster mediates a novel type of intrasteric inhibition and suggest that three events are required for CaMKIV/Gr activation: 1) Ca2+/calmodulin binding; 2) phosphorylation of the Ca2+/calmodulin-bound enzyme on Thr200 by a Ca2+/calmodulin-dependent protein kinase kinase; and 3) autophosphorylation of Ser12-Ser13. This three-step requirement is unique among the multifunctional Ca2+/calmodulin-dependent kinases.

Full Text

Duke Authors

Cited Authors

  • Chatila, T; Anderson, KA; Ho, N; Means, AR

Published Date

  • August 30, 1996

Published In

Volume / Issue

  • 271 / 35

Start / End Page

  • 21542 - 21548

PubMed ID

  • 8702940

International Standard Serial Number (ISSN)

  • 0021-9258

Digital Object Identifier (DOI)

  • 10.1074/jbc.271.35.21542


  • eng

Conference Location

  • United States