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

19 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

Fingerprint

COUP Transcription Factor I
Transcription
Tretinoin
Neuroblastoma
Cells
Cell Line
Assays
Transcription Factors
Messenger RNA
Small Interfering RNA
Genes
ceramide kinase
Co-Repressor Proteins
Sphingolipids
Chromatin Immunoprecipitation
Ceramides
Cell growth
Electrophoretic Mobility Shift Assay
Electrophoresis
Metabolism

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Murakami, Masashi ; Ito, Hiromi ; Hagiwara, Kazumi ; Yoshida, Kayo ; Sobue, Sayaka ; Ichihara, Masatoshi ; Takagi, Akira ; Kojima, Tetsuhito ; Tanaka, Kouji ; Tamiya-Koizumi, Keiko ; Kyogashima, Mamoru ; Suzuki, Motoshi ; Banno, Yoshiko ; Nozawa, Yoshinori ; Murate, Takashi. / ATRA inhibits ceramide kinase transcription in a human neuroblastoma cell line, SH-SY5Y cells : The role of COUP-TFI. In: Journal of Neurochemistry. 2010 ; Vol. 112, No. 2. pp. 511-520.
@article{bba2a8235b214f4a99ccf79260a41df2,
title = "ATRA inhibits ceramide kinase transcription in a human neuroblastoma cell line, SH-SY5Y cells: The role of COUP-TFI",
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.",
author = "Masashi Murakami and Hiromi Ito and Kazumi Hagiwara and Kayo Yoshida and Sayaka Sobue and Masatoshi Ichihara and Akira Takagi and Tetsuhito Kojima and Kouji Tanaka and Keiko Tamiya-Koizumi and Mamoru Kyogashima and Motoshi Suzuki and Yoshiko Banno and Yoshinori Nozawa and Takashi Murate",
year = "2010",
month = "1",
day = "1",
doi = "10.1111/j.1471-4159.2009.06486.x",
language = "English",
volume = "112",
pages = "511--520",
journal = "Journal of Neurochemistry",
issn = "0022-3042",
publisher = "Wiley-Blackwell",
number = "2",

}

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, vol. 112, no. 2, pp. 511-520. https://doi.org/10.1111/j.1471-4159.2009.06486.x

ATRA inhibits ceramide kinase transcription in a human neuroblastoma cell line, SH-SY5Y cells : The role of COUP-TFI. / Murakami, Masashi; Ito, Hiromi; Hagiwara, Kazumi; Yoshida, Kayo; Sobue, Sayaka; Ichihara, Masatoshi; Takagi, Akira; Kojima, Tetsuhito; Tanaka, Kouji; Tamiya-Koizumi, Keiko; Kyogashima, Mamoru; Suzuki, Motoshi; Banno, Yoshiko; Nozawa, Yoshinori; Murate, Takashi.

In: Journal of Neurochemistry, Vol. 112, No. 2, 01.01.2010, p. 511-520.

Research output: Contribution to journalArticle

TY - JOUR

T1 - ATRA inhibits ceramide kinase transcription in a human neuroblastoma cell line, SH-SY5Y cells

T2 - The role of COUP-TFI

AU - Murakami, Masashi

AU - Ito, Hiromi

AU - Hagiwara, Kazumi

AU - Yoshida, Kayo

AU - Sobue, Sayaka

AU - Ichihara, Masatoshi

AU - Takagi, Akira

AU - Kojima, Tetsuhito

AU - Tanaka, Kouji

AU - Tamiya-Koizumi, Keiko

AU - Kyogashima, Mamoru

AU - Suzuki, Motoshi

AU - Banno, Yoshiko

AU - Nozawa, Yoshinori

AU - Murate, Takashi

PY - 2010/1/1

Y1 - 2010/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=72849142996&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=72849142996&partnerID=8YFLogxK

U2 - 10.1111/j.1471-4159.2009.06486.x

DO - 10.1111/j.1471-4159.2009.06486.x

M3 - Article

C2 - 19903244

AN - SCOPUS:72849142996

VL - 112

SP - 511

EP - 520

JO - Journal of Neurochemistry

JF - Journal of Neurochemistry

SN - 0022-3042

IS - 2

ER -