In this study, we examined the effects of GSL deficiency on SRE-mediated gene transcription. For this purpose, we employed GSL-defective melanoma mutant SCH527123 [9], GM95 cells. GM95 is deficient in ceramide glucosyltransferase (GlcT-1), which catalyzes the formation of glucosylceramide, the core structure of major GSLs. Present results showed that the content of mature SREBP is increased in the GSL-defective cells. Transcriptional activation of SREBP target genes and cholesterol synthesis are also induced in the GSL-defective cells. These results indicate that GSL deficiency up-regulates the SREBP pathway.
Materials and methods
Materials. [14C] Serine (165 mCi/mmol) was from Perkin-Elmer (Boston, MA). [1-14C]Acetic acid (54.7 mCi/ml) was purchased from American Radiolabeled Chemicals Inc. (St. Louis, MO). Lipoprotein-deficient serum (LPDS), cholesterol, 25-hydroxycholesterol, lovastatin (mevinolin), and mevalonate were from Sigma (St. Louis, MO). Anti-SREBP2 antibody was kindly provided by Dr. T. Hamakubo of Tokyo University.
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