POLYMORPHIC VARIANTS IN HUMAN MUTY

人类突变体的多态性变异

• 批准号: 6372416
• 负责人: AMANDA K MCCULLOUGH
• 金额: $ 22.12万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6372416
• 关键词: DNA damage; DNA repair; Escherichia coli; HeLa cells; N glycosidase; cell population study; enzyme activity; enzyme structure; genetic polymorphism; genetic screening; human tissue; nucleic acid sequence; oxidative stress; polymerase chain reaction; protein localization; protein purification; protein structure function; site directed mutagenesis

DESCRIPTION (Adapted From the Investigator's Abstract) Recent studies have demonstrated that genetic polymorphisms occur within the genes that comprise the major DNA repair pathways. Mutations within many of these genes can lead to increased toxicity and mutagenicity. Within the base excision repair pathway, one of the primary DNA lesions that is repaired is an oxidatively damaged guanine, 8-oxoG. The accumulation of this lesion has been correlated with aging, cancer, neurodegeneration, and atherosclerosis. One of the genes that is central to the efficient repair of this mismatch is the adenine-specific DNA glycosylase, MutY, that removes adenine at A:8-oxoG mispairs. The human MutY homolog, hMYH, has been cloned and potential polymorphic variants identified. In order to determine the functional significance of these and additional polymorphic variants in hMYH, three independent laboratories have come together with diverse but complementary expertise, to address this question. The first specific aim is to identify additional polymorphic variants in the hMYE gene and these studies will be carried out in the laboratory of H. Mohrenweiser. The choice of samples will be a subset of those that have been preselected by NIH as being representative of the U.S. population. All 16 exons and intron-exon junctions will be analyzed from 96 genomic DNA samples. Specific aim 2 focuses on the structure-function analysis of native hMYH and its polymorphic variants. These studies will be guided and implemented in the laboratory of S. Lloyd, based on the investigator's biochemical and x-ray crystallographic determination of the prokaryotic MutY. The third specific aim, carried out in the laboratory of I. Boldogh, will ascertain the role that cellular oxidative stress has on the regulation and intracellular localization of hMYH. Polymorphic variants that are not affected adversely, with respect to catalysis, may be impaired in stabilization and localization. These goals will be accomplished using a multidisciplinary collaborative approach utilizing expertise in DNA repair enzymology, structure-function analysis, polymorphic screening, and cell biology.

描述（改编自调查员的摘要） 最近的研究表明，遗传多态性发生在 包括主要DNA修复途径的基因。 许多 这些基因可能导致毒性和诱变性增加。 在基础内 切除修复途径，修复的主要DNA病变之一是 氧化受损的鸟嘌呤，8-oxog。 这种病变的积累已经 与衰老，癌症，神经变性和动脉粥样硬化有关。 之一 与此不匹配有效修复至关重要的基因是 腺嘌呤特异性的DNA糖基酶，muty，在A：8-oxog上除去腺嘌呤 失误。 人类的混淆同源物HMYH已被克隆并潜在 确定的多态性变体。 为了确定功能 这些以及HMYH中的其他多态性变体的重要性，三个 独立的实验室已经与多样化但互补的 专业知识，以解决这个问题。 第一个具体目的是确定 HMYE基因中的其他多态性变体，这些研究将是 在H. Mohrenweiser的实验室中进行。 样品的选择将 成为NIH预选为代表的子集 美国人口。 所有16个外显子和内含子 - 外观连接将是 从96个基因组DNA样品中分析。 特定目标2专注于 天然HMYH及其多态性变体的结构 - 功能分析。 这些研究将在S. Lloyd的实验室中进行指导和实施 基于研究者的生化和X射线晶体学 原核生物的测定。 第三个特定目标，在 I. boldogh的实验室将确定细胞氧化的作用 应力对HMYH的调节和细胞内定位具有压力。 相对于没有受到不利影响的多态性变体 催化，在稳定和定位中可能受到损害。 这些目标 将使用多学科协作方法来完成 利用DNA修复酶学的专业知识，结构功能分析， 多态性筛选和细胞生物学。