Imprinting studies of a paternally expressed Usp29 gene

父系表达的 Usp29 基因的印记研究

• 批准号: 6727446
• 负责人: JOOMYEONG KIM
• 金额: $ 8.27万
• 依托单位: UNIVERSITY OF CALIF-LAWRNC LVRMR NAT LAB
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6727446
• 关键词: DNA methylation; artificial chromosomes; cell line; functional /structural genomics; gene deletion mutation; gene expression; genetic enhancer element; genetic mapping; genetic regulation; genetically modified animals; genomic imprinting; laboratory mouse; transcription factor

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.

描述（由申请人提供）：由于驻留基因不寻常的功能半合性，印记区域已与几种不同类型的人类遗传缺陷相关，从神经缺陷到癌症。我们已经确定了位于人类染色体 19q13.4 (HSA19q13.4) 上的一个印记结构域，该区间也与人类和小鼠的印记相关遗传疾病有关。该结构域包含至少 6 个印记基因，包括 Peg3、Usp29、Zim3/Usp29-as、Zim1、Zim2 和 Zfp264。与其他印记域一样，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内每个基因的印记表达是如何受到调节的；拟议的项目特别关注两个父系表达的保守基因 Peg3 和 Usp29 的调控。比较基因组研究使我们能够识别一个差异甲基化区域，我们认为该区域可以作为该间隔的印记控制区域（ICR）。这种潜在的 ICR 与 Peg3 和 Usp29 的启动子区域重叠，称为 P1-DMR（启动子 1 差异甲基化区域）。我们还鉴定了进化上保守的 Gli 型锌指基因 YY1 作为 P1-DMR 的甲基化敏感反式因子。我们预测 P1-DMR/YY1 可能充当甲基化敏感的染色体绝缘子或神经增强子，用于邻近基因的印记控制。为了研究 P1-DMR 对于印记控制的潜在功能，我们将使用几种基于细胞系的测定系统分析从 P1-DMR 观察到的绝缘体和启动子活性。将产生携带 P1-DMR 靶向缺失的突变小鼠来测试 P1-DMR 的体内功能，并且还将使用继承各种报告基因构建体的转基因小鼠研究 YY1 在 P1-DMR 功能中的潜在作用。这些结果将为该印记区域的调控提供新的见解，并探索YY1作为哺乳动物基因组印记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记相关表型提供新线索。