Mechanisms of Genomic Imprinting

描述（由申请人提供）：印记基因表现出体细胞亲本特异性单等位基因表达。印迹基因表达的表观遗传决定因素还没有完全理解，不能仅仅基于CpG甲基化来解释-尚不清楚是什么机制在印迹控制区启动DNA甲基化。印迹基因的染色质差异，如等位基因特异性体内蛋白因子结合和/或组蛋白共价修饰，与体细胞中大量印迹基因的细胞记忆有关，并构成了一层细胞记忆，但在雄性或雌性生殖细胞中，不存在这种差异或它们在印迹控制区的作用的体内证据。这些差异可能存在于性腺特异性DNA甲基化之前，因此可能是建立印记所必需的。这种染色质组成中预先存在的“初级染色质差异”只能通过在获得甲基化印记的时间窗口分析生殖细胞来揭示。我们的研究将验证这一假设，即男性和女性生殖细胞之间存在“初级染色质差异”，并且可能是在印记控制区域建立DNA甲基化印记所必需的。我们将分析正常和突变体体细胞以及雄性和雌性生殖细胞发育过程中H19/lgf 2印迹控制区的染色质结构，以揭示“初级染色质差异”并确认这些“初级染色质差异”在印迹建立中的作用。这些研究与人类健康有关，因为基因组印记是许多人类疾病的基础，如Beckwith-Wiedemann综合征和Wilm肿瘤。基因组印迹的定义在一个基本的水平应该进一步阐明与印迹失败相关的人类疾病的遗传和表观遗传基础。

DESCRIPTION (provided by applicant): Imprinted genes exhibit somatic parental specific monoallelic expression. The epigenetic determinants of imprinted gene expression are not fully understood and cannot be solely explained based on CpG methylation-it is not known what mechanism initiates DNA methylation at the imprinting control regions. Chromatin differences at imprinted genes, such as allele-specific \n vivo protein factor binding and/or histone covalent modifications are associated with-and constitute a layer of-cell memory for a number of imprinted genes in somatic cells, but no in vivo evidence exists of such differences or their role at the imprinting control regions in male or female germ-cells. These differences may exist before gonad-specific DNA methylation, and may, therefore, be required for imprint establishment. Such preexisting "primary chromatin differences" in chromatin composition can only be revealed by the analysis of germ cells at the window of time when methylation imprints are attained. Our studies will test the hypothesis, that "primary chromatin differences" between male and female germ cells exist and may be required for DNA methylation imprint establishment at the imprinting control regions. We will analyze the chromatin structure of the H19/lgf2 imprinting control region in normal and mutant somatic cells and also during male and female germ cell development in order to reveal the "primary chromatin difference(s) and confirm the role of these "primary chromatin differences" in imprint establishment. These studies are related to human health in that genomic imprinting underlies a number of human diseases, such as Beckwith-Wiedemann syndrome and Wilm's tumor. Definition of genomic imprinting at a fundamental level should shed further light on the genetic and epigenetic basis of human diseases associated with imprinting failure.