Insulin Gene Expression Is Regulated by DNA Methylation

被引:230
作者
Kuroda, Akio
Rauch, Tibor A.
Todorov, Ivan
Ku, Hsun Teresa
Al-Abdullah, Ismail H.
Kandeel, Fouad
Mullen, Yoko
Pfeifer, Gerd P.
Ferreri, Kevin
机构
[1] Department of Diabetes, Endocrinology and Metabolism, Research Institute of City of Hope, Duarte, CA
[2] Department of Biology, Beckman Research Institute of City of Hope, Duarte, CA
[3] Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita-City
[4] Department of Orthopedic Surgery, Rush University Medical Center, Chicago, IL
关键词
PROINSULIN BIOSYNTHESIS; BETA-CELLS; BINDING PROTEIN; INS2; LOCUS; CHROMATIN; TRANSCRIPTION; ACTIVATION; PROMOTER; GLUCOSE; 5-METHYLCYTOSINE;
D O I
10.1371/journal.pone.0006953
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background: Insulin is a critical component of metabolic control, and as such, insulin gene expression has been the focus of extensive study. DNA sequences that regulate transcription of the insulin gene and the majority of regulatory factors have already been identified. However, only recently have other components of insulin gene expression been investigated, and in this study we examine the role of DNA methylation in the regulation of mouse and human insulin gene expression. Methodology/Principal Findings: Genomic DNA samples from several tissues were bisulfite-treated and sequenced which revealed that cytosine-guanosine dinucleotide (CpG) sites in both the mouse Ins2 and human INS promoters are uniquely demethylated in insulin-producing pancreatic beta cells. Methylation of these CpG sites suppressed insulin promoter-driven reporter gene activity by almost 90% and specific methylation of the CpG site in the cAMP responsive element (CRE) in the promoter alone suppressed insulin promoter activity by 50%. Methylation did not directly inhibit factor binding to the CRE in vitro, but inhibited ATF2 and CREB binding in vivo and conversely increased the binding of methyl CpG binding protein 2 (MeCP2). Examination of the Ins2 gene in mouse embryonic stem cell cultures revealed that it is fully methylated and becomes demethylated as the cells differentiate into insulin-expressing cells in vitro. Conclusions/Significance: Our findings suggest that insulin promoter CpG demethylation may play a crucial role in beta cell maturation and tissue-specific insulin gene expression.
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页数:9
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