Hypermethylated promoter region of DR3, the death receptor 3 gene, in rheumatoid arthritis synovial cells

Nozomi Takami, Kayo Osawa, Yasushi Miura, Koichiro Komai, Mariko Ikeda, Masahiko Shiraishi, Keizo Sato, Tetsuhiro Iguchi, Kazuko Shiozawa, Akira Hashiramoto, Shunichi Shiozawa

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

Objective. To examine the promoter activity and protein expression of the death receptor 3 gene DR3, a member of the apoptosis-inducing Fas gene family, with particular reference to the methylation status of its promoter region in rheumatoid arthritis (RA). Methods. Genomic DNA was prepared from peripheral blood mononuclear cells obtained from healthy individuals and from patients with RA and synovial cells obtained from patients with MA and osteoarthritis. The methylation status of the DR3 promoter was analyzed by bisulfite genomic sequencing and methylation-specific polymerase chain reaction techniques. Gene promoter activity and protein expression were examined using the luciferase reporter and Western blotting techniques. Results. The promoter region of the DR3 gene contained many CpG motifs, including one CpG island that was specifically hypermethylated in synovial cells from patients with RA. Promoter assays showed that the promoter CpG island was essential for the transactivation of the DR3 gene and that forced hypermethylation of the CpG island with the bacterial methylase Sss I in vitro resulted in inhibition of the DR3 gene expression. Furthermore, the expression of DR-3 protein was down-modulated in association with methylation of the promoter CpG island in MA synovial cells. Conclusion. The CpG island in the DR3 gene promoter was specifically methylated to down-modulate the expression of DR-3 protein in rheumatoid synovial cells, which may provide resistance to apoptosis in RA synovial cells.

Original languageEnglish
Pages (from-to)779-787
Number of pages9
JournalArthritis and Rheumatism
Volume54
Issue number3
DOIs
Publication statusPublished - 01-03-2006

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Receptors, Tumor Necrosis Factor, Member 25
CpG Islands
Genetic Promoter Regions
Rheumatoid Arthritis
Methylation
Genes
Proteins
Apoptosis
Luciferases
Osteoarthritis
Transcriptional Activation
Blood Cells
Western Blotting
Gene Expression
Polymerase Chain Reaction
DNA

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Rheumatology
  • Immunology
  • Pharmacology (medical)

Cite this

Takami, Nozomi ; Osawa, Kayo ; Miura, Yasushi ; Komai, Koichiro ; Ikeda, Mariko ; Shiraishi, Masahiko ; Sato, Keizo ; Iguchi, Tetsuhiro ; Shiozawa, Kazuko ; Hashiramoto, Akira ; Shiozawa, Shunichi. / Hypermethylated promoter region of DR3, the death receptor 3 gene, in rheumatoid arthritis synovial cells. In: Arthritis and Rheumatism. 2006 ; Vol. 54, No. 3. pp. 779-787.
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abstract = "Objective. To examine the promoter activity and protein expression of the death receptor 3 gene DR3, a member of the apoptosis-inducing Fas gene family, with particular reference to the methylation status of its promoter region in rheumatoid arthritis (RA). Methods. Genomic DNA was prepared from peripheral blood mononuclear cells obtained from healthy individuals and from patients with RA and synovial cells obtained from patients with MA and osteoarthritis. The methylation status of the DR3 promoter was analyzed by bisulfite genomic sequencing and methylation-specific polymerase chain reaction techniques. Gene promoter activity and protein expression were examined using the luciferase reporter and Western blotting techniques. Results. The promoter region of the DR3 gene contained many CpG motifs, including one CpG island that was specifically hypermethylated in synovial cells from patients with RA. Promoter assays showed that the promoter CpG island was essential for the transactivation of the DR3 gene and that forced hypermethylation of the CpG island with the bacterial methylase Sss I in vitro resulted in inhibition of the DR3 gene expression. Furthermore, the expression of DR-3 protein was down-modulated in association with methylation of the promoter CpG island in MA synovial cells. Conclusion. The CpG island in the DR3 gene promoter was specifically methylated to down-modulate the expression of DR-3 protein in rheumatoid synovial cells, which may provide resistance to apoptosis in RA synovial cells.",
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Takami, N, Osawa, K, Miura, Y, Komai, K, Ikeda, M, Shiraishi, M, Sato, K, Iguchi, T, Shiozawa, K, Hashiramoto, A & Shiozawa, S 2006, 'Hypermethylated promoter region of DR3, the death receptor 3 gene, in rheumatoid arthritis synovial cells', Arthritis and Rheumatism, vol. 54, no. 3, pp. 779-787. https://doi.org/10.1002/art.21637

Hypermethylated promoter region of DR3, the death receptor 3 gene, in rheumatoid arthritis synovial cells. / Takami, Nozomi; Osawa, Kayo; Miura, Yasushi; Komai, Koichiro; Ikeda, Mariko; Shiraishi, Masahiko; Sato, Keizo; Iguchi, Tetsuhiro; Shiozawa, Kazuko; Hashiramoto, Akira; Shiozawa, Shunichi.

In: Arthritis and Rheumatism, Vol. 54, No. 3, 01.03.2006, p. 779-787.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Hypermethylated promoter region of DR3, the death receptor 3 gene, in rheumatoid arthritis synovial cells

AU - Takami, Nozomi

AU - Osawa, Kayo

AU - Miura, Yasushi

AU - Komai, Koichiro

AU - Ikeda, Mariko

AU - Shiraishi, Masahiko

AU - Sato, Keizo

AU - Iguchi, Tetsuhiro

AU - Shiozawa, Kazuko

AU - Hashiramoto, Akira

AU - Shiozawa, Shunichi

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Y1 - 2006/3/1

N2 - Objective. To examine the promoter activity and protein expression of the death receptor 3 gene DR3, a member of the apoptosis-inducing Fas gene family, with particular reference to the methylation status of its promoter region in rheumatoid arthritis (RA). Methods. Genomic DNA was prepared from peripheral blood mononuclear cells obtained from healthy individuals and from patients with RA and synovial cells obtained from patients with MA and osteoarthritis. The methylation status of the DR3 promoter was analyzed by bisulfite genomic sequencing and methylation-specific polymerase chain reaction techniques. Gene promoter activity and protein expression were examined using the luciferase reporter and Western blotting techniques. Results. The promoter region of the DR3 gene contained many CpG motifs, including one CpG island that was specifically hypermethylated in synovial cells from patients with RA. Promoter assays showed that the promoter CpG island was essential for the transactivation of the DR3 gene and that forced hypermethylation of the CpG island with the bacterial methylase Sss I in vitro resulted in inhibition of the DR3 gene expression. Furthermore, the expression of DR-3 protein was down-modulated in association with methylation of the promoter CpG island in MA synovial cells. Conclusion. The CpG island in the DR3 gene promoter was specifically methylated to down-modulate the expression of DR-3 protein in rheumatoid synovial cells, which may provide resistance to apoptosis in RA synovial cells.

AB - Objective. To examine the promoter activity and protein expression of the death receptor 3 gene DR3, a member of the apoptosis-inducing Fas gene family, with particular reference to the methylation status of its promoter region in rheumatoid arthritis (RA). Methods. Genomic DNA was prepared from peripheral blood mononuclear cells obtained from healthy individuals and from patients with RA and synovial cells obtained from patients with MA and osteoarthritis. The methylation status of the DR3 promoter was analyzed by bisulfite genomic sequencing and methylation-specific polymerase chain reaction techniques. Gene promoter activity and protein expression were examined using the luciferase reporter and Western blotting techniques. Results. The promoter region of the DR3 gene contained many CpG motifs, including one CpG island that was specifically hypermethylated in synovial cells from patients with RA. Promoter assays showed that the promoter CpG island was essential for the transactivation of the DR3 gene and that forced hypermethylation of the CpG island with the bacterial methylase Sss I in vitro resulted in inhibition of the DR3 gene expression. Furthermore, the expression of DR-3 protein was down-modulated in association with methylation of the promoter CpG island in MA synovial cells. Conclusion. The CpG island in the DR3 gene promoter was specifically methylated to down-modulate the expression of DR-3 protein in rheumatoid synovial cells, which may provide resistance to apoptosis in RA synovial cells.

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