Serine 16 of Oncoprotein 18 Is a Major Cytosolic Target for the Ca2+/calmodulin-dependent kinase-Gr

Eur J Biochem. 1994 Oct 1;225(1):53-60. doi: 10.1111/j.1432-1033.1994.00053.x.

Abstract

Oncoprotein 18 (Op18) is a cytosolic protein that was initially identified due to its up-regulated expression in acute leukemia and its complex pattern of phosphorylation in response to diverse extracellular signals. We have previously identified in vivo phosphorylation sites and some of the protein kinase systems involved. Two distinct proline-directed kinase families phosphorylate Ser25 and Ser38 of Op18 with overlapping but distinct site preference. These two kinase families, mitogen-activated protein (MAP) kinases and cyclin-dependent cdc2 kinases, are involved in receptor-regulated and cell-cycle-regulated phosphorylation events, respectively. During analysis of Op18 phosphorylation in the Jurkat T-cell line, we also found that Ser16 of Op18 is phosphorylated in response to a Ca2+ signal generated by T-cell receptor stimulation or the Ca2+ ionophore ionomycin. As suggested by a previous study, T-cell-receptor-induced phosphorylation events may be mediated by the Ca2+/CaM-dependent protein kinase type Gr (CaM kinase-Gr). The present study shows that activation of this protein kinase correlates with phosphorylation of Ser16 of Op18, and in vitro experiments reveal efficient and selective phosphorylation of this residue. The CaM kinase-Gr is only expressed in certain lymphoid cell lines, and the present study shows that ionomycin-induced phosphorylation of Op18 Ser16 is restricted to cells expressing this protein kinase. Finally, CaM kinase-Gr-dependent in vitro phosphorylation of a crude cellular extract reveals a striking preference of this protein kinase for Op18 compared to other cellular substrates. In conclusion, the results suggest that Ser16 of Op18 is a major cytosolic target for activated CaM kinase-Gr.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • CDC2 Protein Kinase / metabolism
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism*
  • Cell Line
  • DNA Primers
  • Humans
  • Isoenzymes / metabolism*
  • Microtubule Proteins*
  • Molecular Sequence Data
  • Peptide Mapping
  • Phosphoproteins / biosynthesis
  • Phosphoproteins / chemistry
  • Phosphoproteins / metabolism*
  • Polymerase Chain Reaction
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Serine*
  • Stathmin
  • Substrate Specificity
  • Tumor Cells, Cultured

Substances

  • DNA Primers
  • Isoenzymes
  • Microtubule Proteins
  • Phosphoproteins
  • Recombinant Proteins
  • STMN1 protein, human
  • Stathmin
  • Serine
  • Calcium-Calmodulin-Dependent Protein Kinases
  • CDC2 Protein Kinase