MOLECULAR PATHOLOGY OF GENETIC DISORDERS

遗传性疾病的分子病理学

• 批准号: 2216471
• 负责人: AKIRA Y YOSHIDA
• 金额: $ 32.43万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-08-01 至 1997-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2216471
• 关键词: adrenoleukodystrophy; analytical method; chemical fingerprinting; chromosomes; computer graphics /printing; enzyme deficiency; enzyme structure; gene expression; genetic disorder; genetic mapping; genetic markers; genetic promoter element; genome; glucose 6 phosphate dehydrogenase; hemolytic anemia; human population genetics; human tissue; immunoelectrophoresis; inborn metabolism disorder; isozymes; liver; mental disorders; molecular pathology; nucleic acid hybridization; nucleic acid sequence; phosphoglycerate kinase; phosphoglycerate kinase deficiency; protein sequence; protein structure function; pseudogenes; restriction mapping

The objective of the proposed project is to elucidate the molecular mechanism of the gene action in humans through analysis of the protein and genomic structures of normal and variant enzymes. The inherited deficiency of the X-linked glucose-6-phosphate dehydrogenase (G6PD) and the X-lined phosphoglycerate kinase (PGK) are associated with various clinical problems, and these two enzymes will be subjected to examination. In the proposed project, the genomic structure of G6PD, including the 5'-promoter region, will be determined by restriction mapping and nucleotide sequencing. The NH2-terminal structure of a putative nascent G6PD will be elucidated by primer-extension analysis of mRNA. Extensive exploration of the restriction fragment-length polymorphisms of the G6PD locus, and detailed mapping of the "G6PD cluster" locus (i.e., genes for hemophilia A- factor VIII, colorblindness, adrenoleukodystrophy, and fragile X- mental retardation) will be undertaken. The exact molecular lesion of the G6PD variants (i.e., associated with chronic and/or drug- or food-induced hemolytic anemia, those associated with functional abnormalities, the common variants existing in various populations, an over-production variant, and a "null" variant) and that of the PGK variants, associated with severe enzyme deficiency, chronic hemolytic anemia and mental disorders, will be determined by analysis of their genomic structures and by mass spectrometric analysis of their protein structures. In addition to the X-linked G6PD, a G6PD-like locus exists on chromosome 17. This locus will be cloned by screening a human genomic library in the cosmid vector. It will then be examined to determine whether the locus is for a non-functional pseudogene or for a functional gene encoding another G6PD isozyme or other enzyme. The three-dimensional structures of horse and yeast PGK are detailed. The structural/functional relationships of the normal and variant human PGK will be visualized by computer graphics analysis. These studies will extend our knowledge of the genomic structures, gene expressions, and genetic abnormalities of the G6PD and PGK loci. The studies may also provide us with valuable information about the clinically important "G6PD cluster" locus.

拟议项目的目的是阐明 人类基因作用的分子机制通过 分析正常和正常的蛋白质和基因组结构 变体酶。 X连锁的遗传缺陷 葡萄糖-6-磷酸脱氢酶（G6PD）和X衬里 磷酸甘油酸激酶（PGK）与各种 临床问题，这两种酶将受到 考试。 在拟议的项目中，G6PD的基因组结构，包括 5'启动器区域将通过限制映射确定 和核苷酸测序。 A的NH2末端结构 推定的新生G6PD将通过引物扩展阐明 mRNA分析。 对限制的广泛探索 G6PD基因座的片段长度多态性，并详细 “ G6PD簇”基因座的映射（即血友病基因a- 因子VIII，色盲，肾上腺素肌营养不良和脆弱的X- 将进行智力低下）。 确切的分子 G6PD变体的病变（即与慢性和/或相关 药物或食物引起的溶血性贫血，与 功能异常，各种存在的常见变体 种群，过度生产的变体以及“无效”变体）和 与严重酶有关的PGK变体 缺乏症，慢性溶血性贫血和精神障碍，将 通过分析其基因组结构和质量来确定 其蛋白质结构的光谱分析。 除X连锁的G6PD外，还有一个类似G6PD的基因座 17染色体。该基因座将通过筛选人来克隆 宇宙载体中的基因组文库。 然后将检查 确定该基因座是针对非功能性假基因还是 对于编码另一个G6PD同工酶或其他编码的功能基因 酶。 马和酵母PGK的三维结构是 详细的。 正常的结构/功能关系 和变体的人类PGK将通过计算机图形可视化 分析。 这些研究将扩展我们对基因组的了解 结构，基因表达和遗传异常 G6PD和PGK基因座。 研究也可能为我们提供 有关临床上重要的“ G6PD）的宝贵信息 群集”基因座。