FROM YEAST PAS GENES TO PEROXISOME BIOGENESIS DISORDER GENES

从酵母 PAS 基因到过氧化物酶体生物发生障碍基因

• 批准号: 6108286
• 负责人: STEPHEN GOULD
• 金额: $ 32.52万
• 依托单位: HUGO W. MOSER RES INST KENNEDY KRIEGER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 1999-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6108286
• 关键词: fungal genetics; gene complementation; gene expression; gene mutation; genetic disorder; high performance liquid chromatography; human genetic material tag; linkage mapping; membrane transport proteins; mental retardation; molecular cloning; molecular pathology; nucleic acid sequence; peroxisome; protein purification; protein sequence; yeasts

Zellweger syndrome, neonatal adrenoleukodystrophy, infantile Refsum~s disease, hyperpipecolic acidemia, and classical rhizomelic chondrodysplasia punctata are a genetically heterogenous group of lethal inherited diseases collectively referred to as the peroxisome biogenesis disorders (PBD). These diseases occur at a frequency of approximately 1/50,000 live births, can manifest a wide range of phenotypes, and are caused by mutations in any of at least 10 different genes. The only common phenotypes of PBD patients are that they suffer from e=severe mental retardation and exhibit a defect in import of at least one class of peroxisomal protein. Using a yeast genetic system, we and others have identified a number of peroxisome assembly protein (PAS) genes required for normal import of peroxisomal matrix proteins. We recently identified the human gene PXR1 on the basis of its similarity to the yeast Pichia pastoris PAS8 gene. The fact that mutations in PXR1 are responsible for complementation group 2 of the PBD provided the first demonstration that human orthologs of yeast PAS genes are defective in the PBD and proved that yeast PAS sequences can be used to clone human PBD genes. The goal of this project is to identify additional human PBD genes by extension of this homology probing strategy. We propose to expand our current molecular analysis of peroxisome assembly in Pichia pastoris in order to identify all genes involved in proxisomal protein import and peroxisome assembly in this yeast. We will study these new yeast PAS genes, proteins and mutants to gain a better understanding of the peroxisome assembly process and use the sequence of these new PAS genes to determine the role of two candidate human PBD genes (PXAAA1 and PXSH3) in peroxisome assembly and human disease.

齐薇格综合征、新生儿肾上腺脑白质营养不良、婴儿 Refsum~s 疾病、高哌胆酸血症和经典根茎病 点状软骨发育不良是一组遗传异质性的致命疾病 遗传性疾病统称为过氧化物酶体生物发生 疾病（PBD）。 这些疾病的发生频率大约为 1/50,000 活产，可以表现出多种表型，并且 由至少 10 个不同基因中的任何一个突变引起。 唯一的 PBD 患者的常见表型是他们患有 e=严重 精神发育迟滞并表现出至少一类进口缺陷 过氧化物酶体蛋白。 使用酵母遗传系统，我们和其他人已经鉴定了许多 正常输入所需的过氧化物酶体组装蛋白（PAS）基因 过氧化物酶体基质蛋白。 我们最近鉴定出人类基因 PXR1 基于其与酵母毕赤酵母 PAS8 基因的相似性。 PXR1 突变负责互补的事实 PBD 的第 2 组首次证明了人类直系同源物 的酵母 PAS 基因在 PBD 中存在缺陷，并证明酵母 PAS 序列可用于克隆人类PBD基因。 此举的目标 该项目的目的是通过扩展该基因来识别其他人类 PBD 基因 同源性探测策略。 我们建议扩大我们目前的分子 分析毕赤酵母中的过氧化物酶体组装，以确定 所有参与近体蛋白输入和过氧化物酶体组装的基因 在这种酵母中。 我们将研究这些新的酵母 PAS 基因、蛋白质和 突变体以获得对过氧化物酶体组装的更好理解 处理并使用这些新 PAS 基因的序列来确定 两个候选人类 PBD 基因（PXAAA1 和 PXSH3）在过氧化物酶体中的作用 装配和人类疾病。