Association of RACK1 and PKCbeta with the common beta-chain of the IL-5/IL-3/GM-CSF receptor

Oncogene. 1999 Sep 9;18(36):5126-30. doi: 10.1038/sj.onc.1202896.

Abstract

Granulocyte macrophage colony stimulating factor (GM-CSF), interleukin-3 (IL-3) and interleukin-5 (IL-5 belong to a family of cytokines that regulate proliferation, differentiation and function of haematopoietic cells. Their receptor consists of a ligand specific alpha-chain and a signal transducing beta-chain (betac). While, the role of phosphotyrosine residues in the betac as mediators of downstream signalling cascades has been established, little is known about non-phosphotyrosine mediated events. To identify proteins interacting with betac, we screened a yeast two-hybrid library with the intracellular domain of betac. We found that RACK1, a molecule associating with activated PKC, PLCgamma and Src kinases, associated with the membrane proximal region of betac in both yeast two-hybrid, immunoprecipitation and GST-pull-down assays. The association of RACK1 was constitutive, demonstrating no alteration upon cellular stimulation. Furthermore, upon stimulation of cells with IL-5 or PMA, a complex of betac and PKCbeta was found. Together, these findings suggest a novel role for RACK1 as a possible adapter molecule associating with the intracellular domain of cytokine receptors.

MeSH terms

  • Animals
  • Base Sequence
  • COS Cells
  • DNA
  • Isoenzymes / metabolism*
  • Peptides / metabolism*
  • Protein Binding
  • Protein Kinase C / metabolism*
  • Protein Kinase C beta
  • Receptors for Activated C Kinase
  • Receptors, Granulocyte-Macrophage Colony-Stimulating Factor / metabolism*
  • Receptors, Interleukin / metabolism*
  • Receptors, Interleukin-3 / metabolism*
  • Receptors, Interleukin-5

Substances

  • Isoenzymes
  • Peptides
  • Receptors for Activated C Kinase
  • Receptors, Granulocyte-Macrophage Colony-Stimulating Factor
  • Receptors, Interleukin
  • Receptors, Interleukin-3
  • Receptors, Interleukin-5
  • peptide I
  • DNA
  • Protein Kinase C
  • Protein Kinase C beta