Effects of overexpression of GMF on cell growth and redox regulation

R. Lim, A. Zaheer, Jeff Kraakevik, C. J. Darby, W. Llf, Oherley

Research output: Contribution to journalArticle

Abstract

GMF (glia maturation factor) is a 17-kDa protein endogenous to the brain. The protein is normally not secreted by the cells and appears to have important intracellular functions. The protein is rapidly and transiently phosphorylated when a cell is simulated by a phorbol ester. GMF contains many phosphorylation sites and can be phosphorylated by at least four kinases: by PKA at T26 and S82, by PKC at S71, by p90 RSK at T26, and by CKII at S52. In an in vitro assay, PKA-phosphorylated GMF shows a strong inhibition on ERK 1/2 MAP kinase and a strong enhancement on p38 MAP kinase. In order to evaluate the intracellular functions of GMF, we overexpressed GMF in C6 glioma cells using two methods: stable transfection using pcDNA3 plasmid, and transient transfection using replication-defective human adenovirus. With both methods, C6 cells transfected with GMF and overexpressing the protein exhibit a lower saturation density in culture compared to non-transfected or vector alone controls. Transfected cells also exhibit morphological differentiation such as the outgrowth of cell processes. When inoculated into nude mice, transfected cells are less tumorigenic than controls. In tissue culture, transfected cells show a 3.5-fold increase in CuZn-dependent Superoxide dismutase (CuZnSOD) activity, but not in MnSOD activity. Western blot analysis reveals a 3.5-fold increase in CuZnSOD protein, suggesting an enzyme induction.

Original languageEnglish (US)
JournalFASEB Journal
Volume12
Issue number8
StatePublished - 1998
Externally publishedYes

Fingerprint

Glia Maturation Factor
Cell growth
Oxidation-Reduction
cell growth
Growth
mitogen-activated protein kinase
cells
Proteins
Superoxide Dismutase
transfection
proteins
Transfection
Tissue culture
superoxide dismutase
Phosphorylation
Mitogen-Activated Protein Kinase 1
Phorbol Esters
p38 Mitogen-Activated Protein Kinases
Human Adenoviruses
Enzyme Induction

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Lim, R., Zaheer, A., Kraakevik, J., Darby, C. J., Llf, W., & Oherley (1998). Effects of overexpression of GMF on cell growth and redox regulation. FASEB Journal, 12(8).

Effects of overexpression of GMF on cell growth and redox regulation. / Lim, R.; Zaheer, A.; Kraakevik, Jeff; Darby, C. J.; Llf, W.; Oherley.

In: FASEB Journal, Vol. 12, No. 8, 1998.

Research output: Contribution to journalArticle

Lim, R, Zaheer, A, Kraakevik, J, Darby, CJ, Llf, W & Oherley 1998, 'Effects of overexpression of GMF on cell growth and redox regulation', FASEB Journal, vol. 12, no. 8.
Lim, R. ; Zaheer, A. ; Kraakevik, Jeff ; Darby, C. J. ; Llf, W. ; Oherley. / Effects of overexpression of GMF on cell growth and redox regulation. In: FASEB Journal. 1998 ; Vol. 12, No. 8.
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