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Interferon-gamma and lipopolysaccharide induce mouse guanylate-binding protein 3 (mGBP3) expression in the murine
1Department of Neurology, Washington University School of Medicine, St. Louis, MO 63110, USA.
Abstract:
Mouse guanylate-binding protein 3 (mGBP3) is a 71-kDa GTPase which belongs to GTP-binding protein family. The present study showed that the expression of mGBP3 transcript was readily induced in a dose dependent fashion in the macrophage cell line RAW264.7 treated with either interferon-gamma (IFN-gamma) or lipopolysaccharide (LPS). The expression of mGBP3 protein was also apparent by 4 and 6 h after the treatment of cells with IFN-gamma (100 U/ml) or LPS (1 microgram/ml), and remained at plateau for at least 24 h. Cycloheximide (10 micrograms/ml) had no effect on the IFN-gamma- or LPS-induced mGBP3 expression, suggesting that the mGBP3 induction did not require further protein synthesis. Interestingly, a protein kinase C (PKC) inhibitor staurosporine (50 nM) abolished the induction of mGBP3 expression by LPS, but not by IFN-gamma. These findings suggest that mGBP3 may be involved in the macrophage activation process and both IFN-gamma and LPS induce the mGBP3 expression through distinct signal transduction pathways.
Insights
Mouse guanylate-binding protein 3 (mGBP3) expression in macrophages is induced by interferon-gamma and lipopolysaccharide. This induction occurs independently of new protein synthesis and involves distinct signaling pathways.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- Guanylate-binding proteins (GBPs) are a family of GTPases involved in cellular processes.
- Mouse guanylate-binding protein 3 (mGBP3) is a 71-kDa GTPase with a poorly understood role in macrophages.
Purpose of the Study:
- To investigate the regulation of mGBP3 expression in macrophages.
- To elucidate the signaling pathways involved in mGBP3 induction by interferon-gamma (IFN-γ) and lipopolysaccharide (LPS).
Main Methods:
- Macrophage cell line RAW264.7 was treated with IFN-γ or LPS.
- mGBP3 transcript and protein levels were analyzed using quantitative methods.
- The effect of cycloheximide and staurosporine on mGBP3 expression was assessed.
Main Results:
- IFN-γ and LPS dose-dependently induced mGBP3 transcript and protein expression in RAW264.7 cells.
- mGBP3 induction occurred without requiring de novo protein synthesis.
- Staurosporine (a protein kinase C inhibitor) blocked LPS-induced mGBP3 expression but not IFN-γ-induced expression.
Conclusions:
- mGBP3 expression is upregulated in macrophages stimulated with IFN-γ and LPS.
- IFN-γ and LPS induce mGBP3 expression through distinct signal transduction pathways.
- mGBP3 may play a role in macrophage activation.
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