Imprinting studies of a paternally expressed Usp29 gene

父系表达的 Usp29 基因的印记研究

• 批准号: 8134853
• 负责人: JOOMYEONG KIM
• 金额: $ 29.67万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 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 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8134853
• 关键词: 19q13.4; Alleles; Binding Sites; Chromosomes, Human, Pair 7; DNA; DNA Methylation; Disease; Embryonic Development; Funding; Gametogenesis; Genes; Genetic Transcription; Genomic Imprinting; Glioma; Global Change; Growth; Hereditary Disease; Human; Human Chromosomes; Human Genetics; Knockout Mice; Light; Link; Malignant neoplasm of ovary; Modeling; Molecular; Mono-S; Mus; Mutant Strains Mice; Mutate; Oogenesis; Patients; Phenotype; Play; Promoter Regions; Repression; Role; Scheme; Site; Small Interfering RNA; Somatic Cell; Testing; YY1 Transcription Factor; base; dosage; histone modification; imprint; in vivo; insight; malignant breast neoplasm; public health relevance; research study; transmission process

DESCRIPTION (provided by applicant): Project Summary Genomic imprinting is closely associated with human genetic disorders due to its functional hemizygosity. The Peg3 domain on human chromosome 19q13.4/proximal mouse chromosome 7 is also associated with several imprinting-related genetic disorders, including frequent loss of human PEG3 expression in the patients of glioma, breast and ovarian cancers. Thus, the long-term objective of this project is to understand the mechanisms controlling the imprinting of the Peg3 domain. In the previous funding cycle, we predicted that the Peg3 domain is likely controlled by one potential ICR (Imprinting Control Region), the Peg3-DMR (Differentially Methylated Region). We subsequently demonstrated that deletion of the Peg3-DMR has a global impact on the transcription of the Peg3 domain. Paternal and maternal transmission of this targeted deletion also resulted in decreased and increased growth rates of the mouse, respectively, an imprinting phenotype associated with proximal mouse chromosome 7. We also discovered that this potential ICR has a very unusual tandem array of YY1 binding sites, and that reducing the in vivo levels of YY1 protein during oogenesis resulted in target-specific DNA hypomethylation on the Peg3-DMR. Based on these observations, we hypothesize 1) that the imprinting of the Peg3 domain is controlled by the Peg3-DMR and 2) that this control requires YY1 as a main trans factor for targeting de novo DNA methylation during oogenesis. In the current proposal, we will test these hypotheses with the following three aims. Aim1 will further characterize the ICR roles played by the Peg3-DMR using the mutant mice lacking the Peg3-DMR. Aim2 will dissect the exact contributions of YY1 to the observed ICR roles of the Peg3-DMR by generating another KO allele with mutated YY1 sites. Aim3 will investigate potential mechanisms by which YY1 establishes and maintains the allele-specific DNA methylation of the Peg3-DMR. The results from these experiments will provide valuable insights regarding previously unnoticed roles for YY1 in genomic imprinting as well as human genetic disorders linked to the Peg3 domain. PUBLIC HEALTH RELEVANCE: A small number of mammalian genes (less than 200) are not functionally equal between the two parental alleles due to an unusual dosage control mechanism, genomic imprinting. The current proposal is aiming to characterize regulatory mechanisms of genomic imprinting.

描述（由申请人提供）：项目概述基因组印记由于其功能半合子性而与人类遗传疾病密切相关。人染色体19q13.4/近端小鼠7号染色体上的Peg 3结构域也与几种印迹相关的遗传疾病相关，包括神经胶质瘤、乳腺癌和卵巢癌患者中人PEG 3表达的频繁缺失。因此，本项目的长期目标是了解控制Peg 3结构域印迹的机制。在上一个资金周期中，我们预测Peg 3结构域可能由一个潜在的ICR（印记控制区），即Peg 3-DMR（差异甲基化区）控制。我们随后证明了Peg 3-DMR的缺失对Peg 3结构域的转录具有全局影响。这种靶向缺失的父系和母系传播也分别导致小鼠生长速率降低和增加，这是与近端小鼠7号染色体相关的印记表型。我们还发现，这种潜在的ICR具有非常不寻常的YY 1结合位点串联阵列，并且在卵子发生期间降低YY 1蛋白的体内水平导致Peg 3-DMR上的靶特异性DNA低甲基化。基于这些观察结果，我们假设1）Peg 3结构域的印记由Peg 3-DMR控制，2）这种控制需要YY 1作为卵子发生期间靶向从头DNA甲基化的主要反式因子。在本提案中，我们将以以下三个目标来检验这些假设。Aim 1将使用缺乏Peg 3-DMR的突变小鼠进一步表征Peg 3-DMR发挥的ICR作用。Aim 2将通过生成另一个具有突变YY 1位点的KO等位基因，分析YY 1对观察到的Peg 3-DMR ICR作用的确切贡献。Aim 3将研究YY 1建立和维持Peg 3-DMR等位基因特异性DNA甲基化的潜在机制。这些实验的结果将为YY 1在基因组印记中的先前未被注意的作用以及与Peg 3结构域相关的人类遗传疾病提供有价值的见解。 公共卫生关系：少数哺乳动物基因（少于200个）由于不寻常的剂量控制机制，基因组印记，在两个亲本等位基因之间功能不相等。目前的建议是旨在表征基因组印记的调控机制。