DNA methylation and chromatin structure regulate T cell perforin gene expression

Qianjin Lu, Ailing Wu, Donna Ray, Chun Deng, John Attwood, Samir Hanash, Matthew Pipkin, Mathias G Lichtenheld, Bruce Richardson

Research output: Contribution to journalArticle

101 Citations (Scopus)

Abstract

Perforin is a cytotoxic effector molecule expressed in NK cells and a subset of T cells. The mechanisms regulating its expression are incompletely understood. We observed that DNA methylation inhibition could increase perforin expression in T cells, so we examined the methylation pattern and chromatin structure of the human perforin promoter and upstream enhancer in primary CD4+ and CD8+ T cells as well as in an NK cell line that expresses perforin, compared with fibroblasts, which do not express perforin. The entire region was nearly completely unmethylated in the NK cell line and largely methylated in fibroblasts. In contrast, only the core promoter was constitutively unmethylated in primary CD4+ and CD8+ cells, and expression was associated with hypomethylation of an area residing between the upstream enhancer at -1 kb and the distal promoter at -0.3 kb. Treating T cells with the DNA methyltransferase inhibitor 5-azacytidine selectively demethylated this area and increased perforin expression. Selective methylation of this region suppressed promoter function in transfection assays. Finally, perforin expression and hypomethylation were associated with localized sensitivity of the 5′ flank to DNase I digestion, indicating an accessible configuration. These results indicate that DNA methylation and chromatin structure participate in the regulation of perforin expression in T cells.

Original languageEnglish
Pages (from-to)5124-5132
Number of pages9
JournalJournal of Immunology
Volume170
Issue number10
StatePublished - May 15 2003

Fingerprint

Perforin
DNA Methylation
Chromatin
T-Lymphocytes
Gene Expression
Natural Killer Cells
Methylation
Fibroblasts
Azacitidine
Cell Line
Deoxyribonuclease I
Methyltransferases
T-Lymphocyte Subsets
Genetic Promoter Regions
Transfection
Digestion
DNA

ASJC Scopus subject areas

  • Immunology

Cite this

Lu, Q., Wu, A., Ray, D., Deng, C., Attwood, J., Hanash, S., ... Richardson, B. (2003). DNA methylation and chromatin structure regulate T cell perforin gene expression. Journal of Immunology, 170(10), 5124-5132.

DNA methylation and chromatin structure regulate T cell perforin gene expression. / Lu, Qianjin; Wu, Ailing; Ray, Donna; Deng, Chun; Attwood, John; Hanash, Samir; Pipkin, Matthew; Lichtenheld, Mathias G; Richardson, Bruce.

In: Journal of Immunology, Vol. 170, No. 10, 15.05.2003, p. 5124-5132.

Research output: Contribution to journalArticle

Lu, Q, Wu, A, Ray, D, Deng, C, Attwood, J, Hanash, S, Pipkin, M, Lichtenheld, MG & Richardson, B 2003, 'DNA methylation and chromatin structure regulate T cell perforin gene expression', Journal of Immunology, vol. 170, no. 10, pp. 5124-5132.
Lu Q, Wu A, Ray D, Deng C, Attwood J, Hanash S et al. DNA methylation and chromatin structure regulate T cell perforin gene expression. Journal of Immunology. 2003 May 15;170(10):5124-5132.
Lu, Qianjin ; Wu, Ailing ; Ray, Donna ; Deng, Chun ; Attwood, John ; Hanash, Samir ; Pipkin, Matthew ; Lichtenheld, Mathias G ; Richardson, Bruce. / DNA methylation and chromatin structure regulate T cell perforin gene expression. In: Journal of Immunology. 2003 ; Vol. 170, No. 10. pp. 5124-5132.
@article{e570c1043b754304acc6db4482572f72,
title = "DNA methylation and chromatin structure regulate T cell perforin gene expression",
abstract = "Perforin is a cytotoxic effector molecule expressed in NK cells and a subset of T cells. The mechanisms regulating its expression are incompletely understood. We observed that DNA methylation inhibition could increase perforin expression in T cells, so we examined the methylation pattern and chromatin structure of the human perforin promoter and upstream enhancer in primary CD4+ and CD8+ T cells as well as in an NK cell line that expresses perforin, compared with fibroblasts, which do not express perforin. The entire region was nearly completely unmethylated in the NK cell line and largely methylated in fibroblasts. In contrast, only the core promoter was constitutively unmethylated in primary CD4+ and CD8+ cells, and expression was associated with hypomethylation of an area residing between the upstream enhancer at -1 kb and the distal promoter at -0.3 kb. Treating T cells with the DNA methyltransferase inhibitor 5-azacytidine selectively demethylated this area and increased perforin expression. Selective methylation of this region suppressed promoter function in transfection assays. Finally, perforin expression and hypomethylation were associated with localized sensitivity of the 5′ flank to DNase I digestion, indicating an accessible configuration. These results indicate that DNA methylation and chromatin structure participate in the regulation of perforin expression in T cells.",
author = "Qianjin Lu and Ailing Wu and Donna Ray and Chun Deng and John Attwood and Samir Hanash and Matthew Pipkin and Lichtenheld, {Mathias G} and Bruce Richardson",
year = "2003",
month = "5",
day = "15",
language = "English",
volume = "170",
pages = "5124--5132",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "10",

}

TY - JOUR

T1 - DNA methylation and chromatin structure regulate T cell perforin gene expression

AU - Lu, Qianjin

AU - Wu, Ailing

AU - Ray, Donna

AU - Deng, Chun

AU - Attwood, John

AU - Hanash, Samir

AU - Pipkin, Matthew

AU - Lichtenheld, Mathias G

AU - Richardson, Bruce

PY - 2003/5/15

Y1 - 2003/5/15

N2 - Perforin is a cytotoxic effector molecule expressed in NK cells and a subset of T cells. The mechanisms regulating its expression are incompletely understood. We observed that DNA methylation inhibition could increase perforin expression in T cells, so we examined the methylation pattern and chromatin structure of the human perforin promoter and upstream enhancer in primary CD4+ and CD8+ T cells as well as in an NK cell line that expresses perforin, compared with fibroblasts, which do not express perforin. The entire region was nearly completely unmethylated in the NK cell line and largely methylated in fibroblasts. In contrast, only the core promoter was constitutively unmethylated in primary CD4+ and CD8+ cells, and expression was associated with hypomethylation of an area residing between the upstream enhancer at -1 kb and the distal promoter at -0.3 kb. Treating T cells with the DNA methyltransferase inhibitor 5-azacytidine selectively demethylated this area and increased perforin expression. Selective methylation of this region suppressed promoter function in transfection assays. Finally, perforin expression and hypomethylation were associated with localized sensitivity of the 5′ flank to DNase I digestion, indicating an accessible configuration. These results indicate that DNA methylation and chromatin structure participate in the regulation of perforin expression in T cells.

AB - Perforin is a cytotoxic effector molecule expressed in NK cells and a subset of T cells. The mechanisms regulating its expression are incompletely understood. We observed that DNA methylation inhibition could increase perforin expression in T cells, so we examined the methylation pattern and chromatin structure of the human perforin promoter and upstream enhancer in primary CD4+ and CD8+ T cells as well as in an NK cell line that expresses perforin, compared with fibroblasts, which do not express perforin. The entire region was nearly completely unmethylated in the NK cell line and largely methylated in fibroblasts. In contrast, only the core promoter was constitutively unmethylated in primary CD4+ and CD8+ cells, and expression was associated with hypomethylation of an area residing between the upstream enhancer at -1 kb and the distal promoter at -0.3 kb. Treating T cells with the DNA methyltransferase inhibitor 5-azacytidine selectively demethylated this area and increased perforin expression. Selective methylation of this region suppressed promoter function in transfection assays. Finally, perforin expression and hypomethylation were associated with localized sensitivity of the 5′ flank to DNase I digestion, indicating an accessible configuration. These results indicate that DNA methylation and chromatin structure participate in the regulation of perforin expression in T cells.

UR - http://www.scopus.com/inward/record.url?scp=0037884760&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037884760&partnerID=8YFLogxK

M3 - Article

VL - 170

SP - 5124

EP - 5132

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 10

ER -