ATRA inhibits ceramide kinase transcription in a human neuroblastoma cell line, SH-SY5Y cells: The role of COUP-TFI

Masashi Murakami, Hiromi Ito, Kazumi Hagiwara, Kayo Yoshida, Sayaka Sobue, Masatoshi Ichihara, Akira Takagi, Tetsuhito Kojima, Kouji Tanaka, Keiko Tamiya-Koizumi, Mamoru Kyogashima, Motoshi Suzuki, Yoshiko Banno, Yoshinori Nozawa, Takashi Murate

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Ceramide is the central lipid in the sphingolipid metabolism. Ceramide kinase (CERK) and its product, ceramide 1-phosphate, 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. Over-expression and small interfering RNA (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 chicken ovalbumin upstream promoter transcription factor I (COUP-TFI), RARα, and RXR, 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.

Original languageEnglish
Pages (from-to)511-520
Number of pages10
JournalJournal of neurochemistry
Volume112
Issue number2
DOIs
Publication statusPublished - 01-01-2010

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cellular and Molecular Neuroscience

Fingerprint Dive into the research topics of 'ATRA inhibits ceramide kinase transcription in a human neuroblastoma cell line, SH-SY5Y cells: The role of COUP-TFI'. Together they form a unique fingerprint.

  • Cite this

    Murakami, M., Ito, H., Hagiwara, K., Yoshida, K., Sobue, S., Ichihara, M., Takagi, A., Kojima, T., Tanaka, K., Tamiya-Koizumi, K., Kyogashima, M., Suzuki, M., Banno, Y., Nozawa, Y., & Murate, T. (2010). ATRA inhibits ceramide kinase transcription in a human neuroblastoma cell line, SH-SY5Y cells: The role of COUP-TFI. Journal of neurochemistry, 112(2), 511-520. https://doi.org/10.1111/j.1471-4159.2009.06486.x