A mutant protein kinase C that can transform fibroblasts

Nature. 1989 Dec 14;342(6251):807-11. doi: 10.1038/342807a0.

Abstract

Expression of normal protein kinase C (PKC) isoenzymes in fibroblasts has been shown to alter growth regulation but has failed to induce complete transformation of the recipient cells. Here we report on a murine ultraviolet-induced fibrosarcoma cell line which has an unusual PKC subcellular distribution with 87% of the PKC activity associated with the membrane. We have cloned and sequenced the alpha-PKC complementary DNA from ultraviolet-induced-fibrosarcoma cells and from mouse Balb/c brain and found four point mutations in the fibrosarcoma PKC, of which three are in the highly conserved regulatory domain and one is in the conserved region of the catalytic domain. Expression of this mutant alpha-PKC gene in normal Balb/c 3T3 fibroblasts results in a fibrosarcoma-like PKC membrane localization and in cell transformation, as judged by their formation of dense foci, anchorage-independent growth and ability to induce solid tumours when inoculated into nude mice. By contast, transfectants expressing the normal alpha-PKC cDNA do not display a morphology typical of malignant transformed cells and fail to induce tumours in vivo. These findings demonstrate that point mutations in the primary structure of PKC modulate enzyme function and are responsible for inducing oncogenicity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cell Compartmentation
  • Cell Division
  • Cell Line
  • Cell Membrane / enzymology
  • Cell Transformation, Neoplastic* / genetics
  • Cell Transformation, Neoplastic* / pathology
  • Cloning, Molecular
  • Fibrosarcoma / enzymology*
  • Fibrosarcoma / genetics
  • Mice
  • Molecular Sequence Data
  • Neoplasms, Experimental / genetics
  • Phorbol 12,13-Dibutyrate / metabolism
  • Protein Kinase C / genetics*
  • Protein Kinase C / metabolism

Substances

  • Phorbol 12,13-Dibutyrate
  • Protein Kinase C