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. 2003 May 15;170(10):5124-32.
doi: 10.4049/jimmunol.170.10.5124.

DNA methylation and chromatin structure regulate T cell perforin gene expression

Qianjin Lu  1 , Ailing Wu, Donna Ray, Chun Deng, John Attwood, Samir Hanash, Matthew Pipkin, Mathias Lichtenheld, Bruce Richardson
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DNA methylation and chromatin structure regulate T cell perforin gene expression

Qianjin Lu et al. J Immunol. .

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 VSports手机版. 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. .

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