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Related Experiment Videos

Galpha12 requires acylation for its transforming activity

T L Jones1, J S Gutkind

  • 1Metabolic Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA. tlzj@helix.nih.gov

Biochemistry
|April 16, 1998
PubMed
Summary
This summary is machine-generated.

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Palmitoylation is crucial for the oncogenic alpha12 G protein

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Oncology

Background:

  • The alpha subunit of G protein G12 (alpha12) with a Q229L mutation acts as an oncogene.
  • Alpha subunits of G proteins undergo post-translational lipid modifications like palmitoylation and myristoylation.
  • These modifications influence protein function and localization.

Purpose of the Study:

  • To investigate the role of alpha12 palmitoylation in membrane localization and oncogenic transformation.
  • To determine if myristoylation can functionally replace palmitoylation for alpha12 activity.

Main Methods:

  • Stable transfection of NIH 3T3 cells with wild-type and mutant alpha12 constructs (Q229L, C11S, S2G).
  • Radiolabeling with [3H]palmitate and [3H]myristate to assess lipid incorporation.

Related Experiment Videos

  • Analysis of protein localization to the particulate fraction.
  • Assays for transformation efficiency: foci formation, soft agar growth, and growth rate.

    • Main Results:

    • Palmitoylation was confirmed in wild-type and QL alpha12 but absent in C11S mutants.
    • Myristoylation was observed only in S2G mutants.
    • Non-palmitoylated C11S,QL alpha12 failed to transform cells.
    • Myristoylation of the S2G mutant restored transformation efficiency, surpassing that of palmitoylated QL alpha12.

    Conclusions:

    • Palmitoylation is essential for the oncogenic transformation mediated by alpha12.
    • Myristoylation can functionally substitute for palmitoylation, enabling alpha12-driven cell transformation.
    • Lipid modification of alpha12 is critical for its oncogenic potential.