The interferon-gamma-mediated inhibition of lipoprotein lipase gene transcription in macrophages involves casein kinase 2- and phosphoinositide-3-kinase-mediated regulation of transcription factors Sp1 and Sp3.
The mechanisms underlying transcriptional inhibition by interferon-gamma (IFN-gamma) are poorly understood despite the existence of a large number of genes that are regulated in this manner and the key role of this cytokine in inflammatory disorders such as atherosclerosis. We have previously identified a novel mechanism for transcriptional inhibition by IFN-gamma that involves a reduction in the binding of transcription factors Sp1 and Sp3 to regulatory sequences in the lipoprotein lipase (LPL) gene. In the present study, we have investigated the signalling pathways that impact on the IFN-gamma-mediated regulation of Sp1/Sp3 binding and LPL gene transcription in macrophages. The IFN-gamma-mediated inhibition of LPL promoter activity was prevented by expression of dominant negative forms of casein kinase 2 (CK2) and protein kinase B (PKB), a key downstream component of the phosphoinositide-3-kinase (PI3K) pathway. IFN-gamma activated both the catalytic subunits of CK2 without affecting their expression. CK2 interacted with both Sp1 and Sp3 and this association was increased by IFN-gamma. Electrophoretic mobility shift assays showed that a CK2-mediated phosphorylation of either cellular extracts or recombinant Sp1 reduced binding to the regulatory region in the LPL gene. The action of PKB was potentially mediated through mammalian target for rapamycin proteins. Taken together, these results suggest a key role for CK2 and PI3K signalling pathways in the IFN-gamma-mediated inhibition of macrophage LPL gene transcription through the regulation of Sp1/Sp3 binding.
Harris
Sandra M
SM
Cardiff School of Biosciences, Cardiff University, Museum Avenue, Cardiff CF10 3US, UK.
Harvey
Elizabeth J
EJ
Hughes
Timothy R
TR
Ramji
Dipak P
DP
D047390:Casein Kinase II / Q000276:immunology / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D006801:Humans; D047468:Immunoprecipitation; D007371:Interferon-gamma / Q000378:metabolism / Q000494:pharmacology*; D008071:Lipoprotein Lipase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation; D015398:Signal Transduction; D051703:Sp Transcription Factors / Q000378:metabolism*; D016329:Sp1 Transcription Factor / Q000378:metabolism; D051705:Sp3 Transcription Factor / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic
D051703:Sp Transcription Factors; D016329:Sp1 Transcription Factor; D051705:Sp3 Transcription Factor; D007371:Interferon-gamma; D019869:Phosphatidylinositol 3-Kinases; D047390:Casein Kinase II; D008071:Lipoprotein Lipase
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