Peroxisome biogenesis and human peroxisome assembly disorders : Approches to prenatal diagnosis and gene therapy.

过氧化物酶体生物合成和人类过氧化物酶体组装障碍：产前诊断和基因治疗方法。

• 批准号: 08557011
• 负责人: FUJIKI Yukio
• 金额: $ 10.69万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08557011/
• 关键词: peroxisome; peroxisome-biogenesis disorders; CHO cell mutants; peroxin; patient gene analysis; AAA family; membrane protein; protein-protein interaction; トポロジー; 患者解析; RINGフィンガー; 亜鉛イオン; 部位変異体; peroxisome; peroxisome-biogenesis disorders; CHO cell mutants; peroxin; patient gene analysis; AAA family; membrane protein; protein-protein interaction; トポロジー; 患者解析; RINGフィンガー; 亜鉛イオン; 部位変異体

With the support of this Grants-in Aid for Scientific Research 0 8 5 5 7 0 1 1 we obtained two major types of findings :I) Isolation, characterization, and complementation group analysis of novel CHO cell mutants defective in peroxisome biogenesisIn addition to the previously isolated, three complementation groups (CGs) of peroxisome-deficient CHO cell mutants, ZP24, Z65, and ZP92, we isolated ZPlO7 (the same group as Z24), ZP1O5/ZP139, ZPIO9, ZP11O, ZP114, ZP119, ZP124, and ZP126, by the P9OH/UV method. CG analysis by PEX cDNA transfection and/or cell fusion with previously identified CGs of mutant cells, including 12 CGs of fibroblasts derived from patients with peroxisome biogenesis disorders (PBDs), revealed that ZP110, ZP114, and ZP126 are distinct from human CGs. Thus, it is evident that peroxisome assembly requires at least 15 gene-products.II) Cloning of novel peroxin genes By genetic functional complementation assay of newly isolated CHO cell mutants, we cloned several peroxin cDNAs (PEXs), including PEX1, PEX12, and PEX19 for ZP1O7, ZP1O9, and ZP119, respectively. We then delineated that gene mutations in PEX1, PEX12, and PEX19 are responsible for PBDs of CG-I (E), CG-III, and CG-J, respectively. Moreover, we isolated PEX1O and PEX16 by expressed sequence tag (EST) DNA search using yeast genes and demonstrated that inactivation of PEX]O and PEX16 is the genetic cause of CG-VII (B) and CG-IX (D) PBDs, respectively. We also found two isoforms of the peroxin Pex5p (PTS 1 receptor) using CG-ll CHO mutants, ZP1O5 and ZP139. Impaired import of PTS1 proteins in the mutant cells was restored by both, shorter and longer, forms of Pex5p. More strikingly, the longer isoform of Pex5p was found to be involved in import of PTS2 proteins as well.

With the support of this Grants-in Aid for Scientific Research 0 8 5 5 7 0 1 1 we obtained two major types of findings :I) Isolation, characterization, and complementation group analysis of novel CHO cell mutants defective in peroxisome biogenesisIn addition to the previously isolated, three complementation groups (CGs) of peroxisome-deficient CHO cell mutants, ZP24, Z65, and ZP92, we isolated ZPlO7 (the same组通过P9OH/UV方法为Z24），ZP1O5/ZP139，ZPIO9，ZP11O，ZP114，ZP119，ZP124和ZP126。 CG通过PEX cDNA转染和/或细胞融合与先前鉴定的突变细胞的CG分析，包括来自过氧化物酶体生物发生障碍患者（PBD）的12个CG的成纤维细胞（PBD），显示ZP110，ZP114和ZP126与人类CG有所不同。因此，很明显，过氧化物组组装需要至少15个基因产物。II）通过新分离的CHO细胞突变体的遗传功能补体测定法克隆新型过氧蛋白基因，我们将几个过氧蛋白CDNA（PEXS）克隆了，包括PEX1，PEX12，PEX12，和PEX19，用于ZP1O7，ZP1O7，ZP1O9，和ZP1O9，和ZP1O9，和ZP1O9，和ZP1O9，和ZP1O9，和ZP1O。然后，我们描述了PEX1，PEX12和PEX19中的基因突变分别负责CG-I（E），CG-III和CG-J的PBD。此外，我们使用酵母基因通过表达的序列TAG（EST）DNA搜索分离了PEX1O和PEX16，并证明PEX] O和PEX16分别是CG-VII（B）和CG-IX（B）和CG-IX（D）PBD的遗传原因。我们还使用CG-LL CHO突变体ZP1O5和ZP139发现了两种过氧蛋白PEX5P（PTS 1受体）的同工型。 PEX5P的较短和更长的形式恢复了突变细胞中PTS1蛋白的进口受损。更引人注目的是，发现PEX5P的同工型也涉及PTS2蛋白的进口。