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Cell type‐specific regulation of the TGF‐β‐responsive α2(I) collagen gene by CpG methylation
Author(s) -
Yamane Kenichi,
Suzuki Hiroyuki,
Ihn Hironobu,
Kato Mitsuyasu,
Yoshikawa Hirohide,
Tamaki Kunihiko
Publication year - 2005
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.20205
Subject(s) - microbiology and biotechnology , type i collagen , dna methylation , chromatin , gene expression , biology , transfection , methylation , regulation of gene expression , chemistry , gene , biochemistry , endocrinology
The α2(I) collagen gene shows cell type‐specific expression, however, the mechanism behind this specificity remains to be determined. We demonstrate here that transforming growth factor‐beta (TGF‐β)‐mediated induction of α2(I) collagen gene is regulated by DNA methylation in a cell type‐specific manner. Human α2(I) collagen mRNA and type I collagen protein were expressed in normal human fibroblasts (NHF), and also strongly enhanced by TGF‐β; they were not detected in HaCaT, HeLa, or HepG2 cells (termed “collagen‐induction resistant (CIR) cells”) even following stimulation with TGF‐β. On the other hand, the transcriptional activity of exogenously transfected α2(I) collagen promoter was clearly up‐regulated by TGF‐β in the CIR cells as well as in NHF. In the CIR cells, CpG clusters around the transcription start site of the α2(I) collagen gene were heavily methylated, whereas no methylation was detected in NHF. Moreover, α2(I) collagen gene was reactivated in the CIR cells by 5‐Aza‐2‐deoxycytidine (5‐AdC) treatment to some extent. However, demethylation by 5‐AdC was limited and it was unable to recover the TGF‐β responsiveness. In NHF, the α2(I) collagen gene has a Smad3‐accessible chromatin structure and acetylated histones in the promoter regions. By contrast, in the CIR cells, Smad3 failed to bind to the chromatin and histones were not acetylated in this area. Furthermore, in vitro methylation of the reporter gene containing the α2(I) collagen promoter significantly reduced both basal and TGF‐β‐induced enhancement of the transcriptional activity in NHF. Thus, we propose that α2(I) collagen gene provides the first example of the TGF‐β responsive gene whose cell type‐specificity is regulated by CpG methylation. © 2004 Wiley‐Liss, Inc.