cpg methylation of a silent controlling element in the murine avy allele is incomplete and unresponsive to methyl donor supplementationcpg甲基化沉默的控制元件的小鼠艾薇等位基因是不完整的和对甲基供体的补充.pdfVIP

CpG Methylation of a Silent Controlling Element in the Murine Avy Allele Is Incomplete and Unresponsive to Methyl Donor Supplementation 1,3 1,2,3 3 1,3 Jennifer E. Cropley , Catherine M. Suter , Kenneth B. Beckman , David I. K. Martin * 1Victor Chang Cardiac Research Institute, Darlinghurst, New South Wales, Australia, 2 Faculty of Medicine, University of New South Wales, Kensington, New South Wales, Australia, 3 Children’s Hospital Oakland Research Institute, Oakland, California, United States of America Abstract Background: The viable yellow allele of agouti (Avy) is remarkable for its unstable and partially heritable epigenetic state, which produces wide variation in phenotypes of isogenic mice. In the Avy allele an inserted intracisternal A particle (IAP) acts as a controlling element which deregulates expression of agouti by transcription from the LTR of the IAP; the phenotypic state has been linked to CpG methylation of the LTR. Phenotypic variation between Avy mice indicates that the epigenetic state of the IAP is unstable in the germline. Principal Findings: We have made a detailed examination of somatic methylation of the IAP using bisulphite allelic sequencing, and find that the promoter is incompletely methylated even when it is transcriptionally silent. In utero exposure to supplementary methyl donors, which alters the spectrum of Avy phenotypes, does not increase the density of CpG methylation in the silent LTR. Conclusions: Our findings suggest that, contrary to previous supposition, methyl donor supplementation acts through an indirect mechanism to silence Avy. The incomplete cytosine methylation we observe at the somatical