Analysis of Pax2 target genes involved in metanephric kidney morphogenesis

Pax2参与后肾形态发生的靶基因分析

• 批准号: 7683903
• 负责人: Kristopher Robert Schwab
• 金额: $ 5.13万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7683903
• 关键词: Accounting; Acetylation; Affect; Binding; Binding Proteins; Binding Sites; Biochemical Genetics; Cell Aging; Cell Line; Cell Lineage; Cells; Chromatin; Chromatin Structure; Classification; Complex; Cues; DNA-Binding Proteins; Data; Development; Disease; Embryonic Development; Enhancers; Environment; Epigenetic Process; Epithelial; Failure; Family; Gene Activation; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Programming; Genetic Transcription; Genitourinary system; Genome; Genomics; Heterochromatin; Histocompatibility Testing; Histone H3; Histones; Human Genome; Inherited; Kidney; Kidney Diseases; Lysine; Malignant Neoplasms; Mediating; Memory; Mesenchyme; Metanephric structure; Methodology; Methods; Methylation; Methyltransferase; Microarray Analysis; Modeling; Modification; Morphogenesis; Mouse Strains; Organogenesis; Pathway interactions; Pattern; Phosphorylation; Play; Polycystic Kidney Diseases; Process; Processed Genes; Regulator Genes; Role; Site; Specific qualifier value; Specificity; Structure; System; Tail; Technology; Testing; Tissues; Tube; Ubiquitination; Work; cell age; chromatin immunoprecipitation; embryo tissue; genome wide association study; histone modification; imprint; in vivo; link protein; molecular pathology; mouse genome; nephrogenesis; overexpression; progenitor; promoter; research study; response; tissue/cell culture; transcription factor

The human genome contains all of the information to make thousands of specialized cells and tissues during embryonic development. Even though all cells contain the same genes, cellular differentiation requires the selective activation and suppression of genes during development. Failure to correctly regulate this process of gene selectivity results in cancer, developmental abnormalities, cellular degeneration, and many other disease states. Gene activation and suppression patterns during cellular differentiation are specified by epigenetic mechanisms that are heritable and subject to modifications. Eukaryotic chromatin consists of DMA wrapped around a histone octamer. Modification of the core histone tails by acetylation, phosphorylation, methylation or ubiquitination can dramatically alter the local chromatin structure and the potential for gene expression. Accumulated biochemical and genetic evidence indicates that methylation at specific lysine residues of histones H3 and H4 can determine whether a gene remains accessible to the transcription machinery or whether it is silenced into tightly packaged heterochromatin. Such epigentic modifications of histones could account for a heritable cellular memory during embryonic development and could greatly affect gene expression patterns in diseased and aging cells. The Pax2 gene encodes a DNA binding protein and is essential for specifying the kidney and urogenital tract early in development. Loss of Pax2 function results in complete renal agenesis. Yet, overexpression of Pax2 is associated with a variety or renal diseases, including cancer and polycystic kidney disease. Recently, we identified a protein that links Pax2 to a mammalian histone H3 lysine 4 methyltransferase complex. In order to understand how this complex regulates renal patterning and disease, we must identify genes that are directly regulated by Pax2 and determine the sites of Pax2/DNA interactions. This propsal will take a systematic approach to define Pax2 target genes and Pax2 binding sites within the mouse genome. We will demonstrate changes in epigenetic imprinting patterns at these binding sites in response to Pax2 activity. Using both embryonic tissues and cell culture models, we will test the significance of Pax2/DNA interactions and characterize the modifcations of histone methylation at these sites. These experiments will define how a developmental regulatory gene that specifies cell fate can alter chromatin structure in a heritable way.

人类基因组包含制造数千种特殊细胞和组织的所有信息 在胚胎发育过程中。尽管所有细胞都含有相同的基因，但细胞分化 需要在发育过程中选择性激活和抑制基因。未能正确监管 这种基因选择性过程会导致癌症、发育异常、细胞退化和 许多其他疾病状态。细胞分化过程中的基因激活和抑制模式是 由可遗传且可修改的表观遗传机制指定。真核染色质 由包裹着组蛋白八聚体的 DMA 组成。通过乙酰化修饰核心组蛋白尾部， 磷酸化、甲基化或泛素化可以显着改变局部染色质结构和 基因表达的潜力。积累的生化和遗传证据表明，甲基化 组蛋白 H3 和 H4 的特定赖氨酸残基可以确定基因是否仍可被 转录机制或是否被沉默成紧密包装的异染色质。这样的表观遗传 组蛋白的修饰可以解释胚胎发育过程中可遗传的细胞记忆 可以极大地影响患病和衰老细胞的基因表达模式。 Pax2 基因编码 DNA 结合蛋白，对于确定肾脏和泌尿生殖系统至关重要 道处于开发早期。 Pax2 功能丧失会导致肾完全发育不全。然而，过度表达 Pax2与多种肾脏疾病相关，包括癌症和多囊肾病。 最近，我们发现了一种将 Pax2 与哺乳动物组蛋白 H3 赖氨酸 4 甲基转移酶连接的蛋白质 复杂的。为了了解这种复合物如何调节肾脏模式和疾病，我们必须确定 受 Pax2 直接调控并决定 Pax2/DNA 相互作用位点的基因。这个提案 将采取系统方法来定义小鼠体内的 Pax2 靶基因和 Pax2 结合位点 基因组。我们将展示这些结合位点的表观遗传印记模式的变化作为响应 Pax2 活动。使用胚胎组织和细胞培养模型，我们将测试 Pax2/DNA 相互作用并表征这些位点的组蛋白甲基化修饰。这些 实验将确定指定细胞命运的发育调控基因如何改变染色质 以可遗传的方式进行结构。