[Abstract]  [Full Text]  [PDF]

Qianjin Lu^1* and Bruce Richardson¹

¹ Department of Medicine, University of Michigan, the Ann Arbor Veterans Affairs Medical Center. Ann Arbor MI, 48109-0940, USA and the Second Xiangya Hospital, Central South University, Changsha. China.

* To whom correspondence should be addressed: Qianjin Lu, Department of Medicine, University of Michigan, the Ann Arbor Veterans Affairs Medical Center. Ann Arbor MI, 48109-0940, USA and the Second Xiangya Hospital, Central South University, Changsha. China. Email: qianlu@umich.edu

Biol. Proced. Online 2004;6:189-203. doi:10.1251/bpo89
Submitted: August 06, 2004; Accepted: August 24, 2004; Published: September 16, 2004.

Indexing terms: DNA methylation; T-lymphocytes; Gene expression regulation.

Abbreviations: PHA, phytohemagglutinin; 5-azaC, 5-azacytidine; dC, deoxycytosine; SAM, S-adenosylmethionine; MeCP2, methyl-CpG-binding protein 2.

Chromatin structure, determined in part by DNA methylation, is established during differentiation and prevents expression of genes unnecessary for the function of a given cell type. We reported that DNA methylation and chromatin structure contributes to lymphoid-specific ITGAL (CD11a) and PRF1 (perforin) expression. We used bisulfite sequencing to compare methylation patterns in the ITGAL promoter and 5' flanking region of T cells and fibroblasts, and in the PRF1 promoter and upstream enhancer of CD4+ and CD8+ T cells with fibroblasts. The effects of methylation on promoter function were tested using regional methylation of reporter constructs, and confirmed by DNA methyltransferase inhibition. The relationship between DNA methylation and chromatin structure was analyzed by DNaseI hypersensitivity. Herein we described the methods and results in greater detail.