Genomic matrix attachment region and chromosome conformation capture quantitative real time PCR assays identify novel putative regulatory elements at the imprinted Dlk1/Gtl2 locus

被引:26
作者
Braem, Caroline [1 ]
Recolin, Benedicte [1 ]
Rancourt, Rebecca C. [2 ]
Angiolini, Christopher [2 ]
Barthes, Pauline [1 ]
Branchu, Priscillia [1 ]
Court, Franck [1 ]
Cathala, Guy [1 ]
Ferguson-Smith, Anne C. [2 ]
Forne, Thierry [1 ]
机构
[1] Univ Montpellier 2, CNRS, UMR 5535, Inst Genet Mol,IGMM,IFR 122, F-34293 Montpellier 5, France
[2] Univ Cambridge, Dept Physiol Dev & Neurosci, Cambridge CB2 3DY, England
基金
英国医学研究理事会;
关键词
D O I
10.1074/jbc.M801883200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We previously showed that genomic imprinting regulates matrix attachment region activities at the mouse Igf2 (insulin-like growth factor 2) locus and that these activities are functionally linked to neighboring differentially methylated regions (DMRs). Here, we investigate the similarly structured Dlk1/Gtl2 imprinted domain and show that in the mouse liver, the G/C-rich intergenic germ line-derived DMR, a sequence involved in domain-wide imprinting, is highly retained within the nuclear matrix fraction exclusively on the methylated paternal copy, reflecting its differential function on that chromosome. Therefore, not only "classical" A/T-rich matrix attachment region (MAR) sequences but also other important regulatory DNA elements (such as DMRs) can be recovered from genomic MAR assays following a high salt treatment. Interestingly, the recovery of one A/T-rich sequence (MAR4) from the "nuclear matrix" fraction is strongly correlated with gene expression. We show that this element possesses an intrinsic activity that favors transcription, and using chromosome conformation capture quantitative real time PCR assays, we demonstrate that the MAR4 interacts with the intergenic germ line-derived DMR specifically on the paternal allele but not with the Dlk1/Gtl2 promoters. Altogether, our findings shed a new light on gene regulation at this locus.
引用
收藏
页码:18612 / 18620
页数:9
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