Imprinting studies of a paternally expressed Usp29 gene

描述（由申请人提供）：由于常驻基因的不寻常的功能半合子性，印记区域与几种不同类型的人类遗传缺陷有关，从神经缺陷到癌症。我们已经确定了一个位于人类染色体19q13.4的印迹结构域（HSA19q13.4），该区间也与人类和小鼠的印迹相关遗传疾病有关。该结构域包含至少6个印迹基因，包括Peg 3、Usp 29、Zim 3/Usp 29-as、Zim 1、Zim 2和Zfp 264。与其他印迹域一样，基因聚类被认为反映了远程控制机制的存在。这项工作的长期目标是了解该域中每个基因的印记表达是如何调节的;拟议的项目特别关注两个父系表达的保守基因Peg 3和Usp 29的调节。比较基因组学研究使我们能够确定一个差异甲基化的区域，我们认为这可能是一个印记控制区（ICR）的这段时间。这种潜在的ICR与Peg 3和Usp 29的启动子区域重叠，称为P1-DMR（启动子1-差异甲基化区域）。我们还确定了一个进化上保守的Gli型锌指基因YY 1作为P1-DMR的甲基化敏感的反式因子。我们预测P1-DMR/YY 1可能作为甲基化敏感的染色体绝缘子或神经增强子，用于对邻近基因的印迹控制。为了研究P1-DMR对印迹控制的潜在功能，我们将使用几种基于细胞系的测定系统分析从P1-DMR观察到的绝缘子和启动子活性。将产生携带P1-DMR靶向缺失的突变小鼠以测试P1-DMR的体内功能，并且还将使用遗传各种报告构建体的转基因小鼠研究YY 1在P1-DMR功能中的潜在作用。这些结果将为该印迹区域的调控提供新的见解，并探索YY 1作为哺乳动物基因组印迹的反式因子的可能作用。这些研究将为人类和小鼠中与该结构域相关的印迹相关表型提供新的线索。

DESCRIPTION (provided by applicant): Due to the unusual functional hemizygosity of the resident genes, imprinted regions have been linked to several different kinds of human genetic defects, ranging from neurological defects to cancer. We have characterized one imprinted domain located in human chromosome 19q13.4 (HSA19q13.4), and this interval is also associated with imprinting related genetic disorders in human and mouse. This domain contains at least 6 imprinted genes, including Peg3, Usp29, Zim3/Usp29-as, Zim1, Zim2, and Zfp264. As in other imprinted domains gene clustering is thought to reflect the presence of long-range controlling mechanism(s). The long-term goal of this work is to understand how the imprinted expression of each gene within this domain is regulated; the proposed project is focused specifically on the regulation of two paternally expressed conserved genes, Peg3 and Usp29. Comparative genomic studies allowed us to identify one differentially methylated region that we believe may serve as an Imprinting Control Region (ICR) for this interval. This potential ICR overlaps with the promoter region of Peg3 and Usp29, termed P1-DMR (Promoter 1-Differentially Methylated Region). We have also identified an evolutionarily conserved Gli-type zinc-finger gene YY1 as a methylation-sensitive trans factor for P1-DMR. We predict that P1-DMR/YY1 may function either as a methylation sensitive chromosomal insulator or neural enhancer for the imprinting control of the neighboring genes. To investigate the potential function of P1-DMR for imprinting control, we will analyze the insulator and promoter activity observed from P1-DMR using several cell-line based assay systems. Mutant mice carrying targeted deletions of P1-DMR will be generated to test the in vivo function of P1-DMR, and the potential roles of YY1 in the P1-DMR function will also be investigated using transgenic mice inheriting various reporter constructs. These results will provide new insights into the regulation of this imprinted region, and explore the possible role of YY1 as a trans factor for mammalian genomic imprinting. These studies will provide new clues to imprinting-related phenotypes linked to this domain in human and mouse.