Heterogeneous sphingosine-1-phosphate lyase gene expression and its regulatory mechanism in human lung cancer cell lines

Hiromi Ito, Kayo Yoshida, Masashi Murakami, Kazumi Hagiwara, Noriko Sasaki, Misa Kobayashi, Akira Takagi, Tetsuhito Kojima, Sayaka Sobue, Motoshi Suzuki, Keiko Tamiya-Koizumi, Mitsuhiro Nakamura, Yoshiko Banno, Yoshinori Nozawa, Takashi Murate

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The role of sphingolipid metabolic pathway has been recognized in determining cellular fate. Although sphingolipid degradation has been extensively studied, gene expression of human sphingosine 1-phosphate lyase (SPL) catalyzing sphingosine 1-phosphate (S1P) remains to be determined. Among 5 human lung cancer cell lines examined, SPL protein levels paralleled the respective mRNA and enzyme activities. Between H1155 and H1299 cells used for further experiments, higher cellular S1P was observed in H1155 with higher SPL activity compared with H1299 with low SPL activity. GATA-4 has been reported to affect SPL transcription in Dictyostelium discoideum. GATA-4 was observed in H1155 but not in other cell lines. Overexpression of GATA-4 in H1299 increased SPL expression. However, promoter analysis of human SPL revealed that the most important region was located between - 136 bp and - 88 bp from the first exon, where 2 Sp1 sites exist but no GATA site. DNA pull-down assay of H1155 showed increased DNA binding of Sp1 and GATA-4 within this promoter region compared with H1299. Electrophoresis mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP) assay, reporter assay using mutated binding motif, and mithramycin A, a specific Sp1 inhibitor, suggest the major role of Sp1 in SPL transcription and no direct binding of GATA-4 with this 5′ promoter region. The collaborative role of GATA-4 was proved by showing coimmunoprecipitation of Sp1 and GATA-4 using GST-Sp1 and overexpressed GATA-4. Thus, high SPL transcription of H1155 cells was regulated by Sp1 and GATA-4/Sp1 complex formation, both of which bind to Sp1 sites of the 5′-SPL promoter.

Original languageEnglish
Pages (from-to)119-128
Number of pages10
JournalBiochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
Volume1811
Issue number3
DOIs
Publication statusPublished - 01-03-2011

Fingerprint

Lung Neoplasms
Gene Expression
Cell Line
Sphingolipids
Genetic Promoter Regions
sphingosine 1-phosphate lyase (aldolase)
Dictyostelium
Chromatin Immunoprecipitation
DNA
Electrophoretic Mobility Shift Assay
Metabolic Networks and Pathways
Electrophoresis
Exons
Messenger RNA
Enzymes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Ito, Hiromi ; Yoshida, Kayo ; Murakami, Masashi ; Hagiwara, Kazumi ; Sasaki, Noriko ; Kobayashi, Misa ; Takagi, Akira ; Kojima, Tetsuhito ; Sobue, Sayaka ; Suzuki, Motoshi ; Tamiya-Koizumi, Keiko ; Nakamura, Mitsuhiro ; Banno, Yoshiko ; Nozawa, Yoshinori ; Murate, Takashi. / Heterogeneous sphingosine-1-phosphate lyase gene expression and its regulatory mechanism in human lung cancer cell lines. In: Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids. 2011 ; Vol. 1811, No. 3. pp. 119-128.
@article{36fc98bce2ce40628e2dbbed951ed691,
title = "Heterogeneous sphingosine-1-phosphate lyase gene expression and its regulatory mechanism in human lung cancer cell lines",
abstract = "The role of sphingolipid metabolic pathway has been recognized in determining cellular fate. Although sphingolipid degradation has been extensively studied, gene expression of human sphingosine 1-phosphate lyase (SPL) catalyzing sphingosine 1-phosphate (S1P) remains to be determined. Among 5 human lung cancer cell lines examined, SPL protein levels paralleled the respective mRNA and enzyme activities. Between H1155 and H1299 cells used for further experiments, higher cellular S1P was observed in H1155 with higher SPL activity compared with H1299 with low SPL activity. GATA-4 has been reported to affect SPL transcription in Dictyostelium discoideum. GATA-4 was observed in H1155 but not in other cell lines. Overexpression of GATA-4 in H1299 increased SPL expression. However, promoter analysis of human SPL revealed that the most important region was located between - 136 bp and - 88 bp from the first exon, where 2 Sp1 sites exist but no GATA site. DNA pull-down assay of H1155 showed increased DNA binding of Sp1 and GATA-4 within this promoter region compared with H1299. Electrophoresis mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP) assay, reporter assay using mutated binding motif, and mithramycin A, a specific Sp1 inhibitor, suggest the major role of Sp1 in SPL transcription and no direct binding of GATA-4 with this 5′ promoter region. The collaborative role of GATA-4 was proved by showing coimmunoprecipitation of Sp1 and GATA-4 using GST-Sp1 and overexpressed GATA-4. Thus, high SPL transcription of H1155 cells was regulated by Sp1 and GATA-4/Sp1 complex formation, both of which bind to Sp1 sites of the 5′-SPL promoter.",
author = "Hiromi Ito and Kayo Yoshida and Masashi Murakami and Kazumi Hagiwara and Noriko Sasaki and Misa Kobayashi and Akira Takagi and Tetsuhito Kojima and Sayaka Sobue and Motoshi Suzuki and Keiko Tamiya-Koizumi and Mitsuhiro Nakamura and Yoshiko Banno and Yoshinori Nozawa and Takashi Murate",
year = "2011",
month = "3",
day = "1",
doi = "10.1016/j.bbalip.2010.12.005",
language = "English",
volume = "1811",
pages = "119--128",
journal = "Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids",
issn = "1388-1981",
publisher = "Elsevier",
number = "3",

}

Ito, H, Yoshida, K, Murakami, M, Hagiwara, K, Sasaki, N, Kobayashi, M, Takagi, A, Kojima, T, Sobue, S, Suzuki, M, Tamiya-Koizumi, K, Nakamura, M, Banno, Y, Nozawa, Y & Murate, T 2011, 'Heterogeneous sphingosine-1-phosphate lyase gene expression and its regulatory mechanism in human lung cancer cell lines', Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids, vol. 1811, no. 3, pp. 119-128. https://doi.org/10.1016/j.bbalip.2010.12.005

Heterogeneous sphingosine-1-phosphate lyase gene expression and its regulatory mechanism in human lung cancer cell lines. / Ito, Hiromi; Yoshida, Kayo; Murakami, Masashi; Hagiwara, Kazumi; Sasaki, Noriko; Kobayashi, Misa; Takagi, Akira; Kojima, Tetsuhito; Sobue, Sayaka; Suzuki, Motoshi; Tamiya-Koizumi, Keiko; Nakamura, Mitsuhiro; Banno, Yoshiko; Nozawa, Yoshinori; Murate, Takashi.

In: Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids, Vol. 1811, No. 3, 01.03.2011, p. 119-128.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Heterogeneous sphingosine-1-phosphate lyase gene expression and its regulatory mechanism in human lung cancer cell lines

AU - Ito, Hiromi

AU - Yoshida, Kayo

AU - Murakami, Masashi

AU - Hagiwara, Kazumi

AU - Sasaki, Noriko

AU - Kobayashi, Misa

AU - Takagi, Akira

AU - Kojima, Tetsuhito

AU - Sobue, Sayaka

AU - Suzuki, Motoshi

AU - Tamiya-Koizumi, Keiko

AU - Nakamura, Mitsuhiro

AU - Banno, Yoshiko

AU - Nozawa, Yoshinori

AU - Murate, Takashi

PY - 2011/3/1

Y1 - 2011/3/1

N2 - The role of sphingolipid metabolic pathway has been recognized in determining cellular fate. Although sphingolipid degradation has been extensively studied, gene expression of human sphingosine 1-phosphate lyase (SPL) catalyzing sphingosine 1-phosphate (S1P) remains to be determined. Among 5 human lung cancer cell lines examined, SPL protein levels paralleled the respective mRNA and enzyme activities. Between H1155 and H1299 cells used for further experiments, higher cellular S1P was observed in H1155 with higher SPL activity compared with H1299 with low SPL activity. GATA-4 has been reported to affect SPL transcription in Dictyostelium discoideum. GATA-4 was observed in H1155 but not in other cell lines. Overexpression of GATA-4 in H1299 increased SPL expression. However, promoter analysis of human SPL revealed that the most important region was located between - 136 bp and - 88 bp from the first exon, where 2 Sp1 sites exist but no GATA site. DNA pull-down assay of H1155 showed increased DNA binding of Sp1 and GATA-4 within this promoter region compared with H1299. Electrophoresis mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP) assay, reporter assay using mutated binding motif, and mithramycin A, a specific Sp1 inhibitor, suggest the major role of Sp1 in SPL transcription and no direct binding of GATA-4 with this 5′ promoter region. The collaborative role of GATA-4 was proved by showing coimmunoprecipitation of Sp1 and GATA-4 using GST-Sp1 and overexpressed GATA-4. Thus, high SPL transcription of H1155 cells was regulated by Sp1 and GATA-4/Sp1 complex formation, both of which bind to Sp1 sites of the 5′-SPL promoter.

AB - The role of sphingolipid metabolic pathway has been recognized in determining cellular fate. Although sphingolipid degradation has been extensively studied, gene expression of human sphingosine 1-phosphate lyase (SPL) catalyzing sphingosine 1-phosphate (S1P) remains to be determined. Among 5 human lung cancer cell lines examined, SPL protein levels paralleled the respective mRNA and enzyme activities. Between H1155 and H1299 cells used for further experiments, higher cellular S1P was observed in H1155 with higher SPL activity compared with H1299 with low SPL activity. GATA-4 has been reported to affect SPL transcription in Dictyostelium discoideum. GATA-4 was observed in H1155 but not in other cell lines. Overexpression of GATA-4 in H1299 increased SPL expression. However, promoter analysis of human SPL revealed that the most important region was located between - 136 bp and - 88 bp from the first exon, where 2 Sp1 sites exist but no GATA site. DNA pull-down assay of H1155 showed increased DNA binding of Sp1 and GATA-4 within this promoter region compared with H1299. Electrophoresis mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP) assay, reporter assay using mutated binding motif, and mithramycin A, a specific Sp1 inhibitor, suggest the major role of Sp1 in SPL transcription and no direct binding of GATA-4 with this 5′ promoter region. The collaborative role of GATA-4 was proved by showing coimmunoprecipitation of Sp1 and GATA-4 using GST-Sp1 and overexpressed GATA-4. Thus, high SPL transcription of H1155 cells was regulated by Sp1 and GATA-4/Sp1 complex formation, both of which bind to Sp1 sites of the 5′-SPL promoter.

UR - http://www.scopus.com/inward/record.url?scp=78651228625&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78651228625&partnerID=8YFLogxK

U2 - 10.1016/j.bbalip.2010.12.005

DO - 10.1016/j.bbalip.2010.12.005

M3 - Article

C2 - 21184844

AN - SCOPUS:78651228625

VL - 1811

SP - 119

EP - 128

JO - Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids

JF - Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids

SN - 1388-1981

IS - 3

ER -