@phdthesis{oai:nagoya.repo.nii.ac.jp:00012550, author = {村上, 真史 and MURAKAMI, Masashi}, month = {Mar}, note = {Ceramide is the central lipid in the sphingolipid metabolism. Ceramide kinase (CERK) and its product, ceramide 1-phosphate (C1P), have been implicated in various cellular functions. However, the regulatory mechanism of CERK gene expression remains to be determined. Here, we examined CERK mRNA level during all-trans retinoic acid (ATRA)-induced differentiation of a human neuroblastoma cell line, SH-SY5Y. ATRA reduced CERK mRNA and protein levels. Overexpression and siRNA of CERK revealed that CERK is inhibitory against ATRA-induced neuronal differentiation and cell growth arrest. ATRA inhibited the transcriptional activity of 5’-promoter of CERK. Truncation and mutation study suggests that ATRA-responsible region was mainly located in the tandem retinoic acid responsive elements (RARE) between -40 bp and the first exon. The electrophoresis mobility shift assay revealed that ATRA produced two retarded bands, which were erased by antibody against COUP-TFI, RARα and RXRα, respectively. DNA pull-down assay confirmed increased binding of these transcription factors to RARE. Transient expression of RAR, RXR and COUP-TFI and siRNA transfection of these genes revealed that COUP-TFI inhibited CERK mRNA. Furthermore, chromatin immunoprecipitation assay showed the recruitment of co-repressors as well as three transcription factors. These results suggest that COUP-TFI was the ATRA-responsive suppressive transcription factor of CERK gene transcription., 名古屋大学博士学位論文 学位の種類:博士(医療技術学) (課程) 学位授与年月日 平成22年3月25日}, school = {名古屋大学, NAGOYA University}, title = {ATRA inhibits ceramide kinase transcription through an ATRA-related transcription factor, COUP-TFI, in a human neuroblastoma cell line, SH-SY5Y}, year = {2010} }