|Budget Amount *help
¥30,800,000 (Direct Cost: ¥30,800,000)
Fiscal Year 1997: ¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 1996: ¥8,100,000 (Direct Cost: ¥8,100,000)
Fiscal Year 1995: ¥15,300,000 (Direct Cost: ¥15,300,000)
I.Isolation, characterization, and complementation group analysis of novel CHO cell mutants defective in peroxisome biogenesis.
In addition to previously isolated, three complementation groups (CG_S) of peroxisome-deficient CHO cell mutants, ZP24, Z65, and ZP92, we isolated ZP107 (the same group as Z24), ZP105/ZP139, ZP109, ZP110.ZP114, and ZP119, by the P9OH/UV method. CG analysis by PEX cDNA transfection and/or cell fusion with previously identified CGs of mutant cells, including 10 groups of fibroblasts derived from patients with peroxisomal disorders, revealed ZP110, ZP114, and ZP119 to be in 3 novel CGs. Thus, it is evident that peroxisome assembly requires at least 13 genes products.
II.Cloning of PEX12 and PEX1
By genetic functional complementation assay, PEX12 and PEX1 were cloned for ZP109 and ZP107, respectively. Mutation analysis of PEX12 and PEX1 in patients with Zellweger syndorome of CGs III and I,respectively, were also done. Inactivation of PEX12 and PEX1 was demonstrated to be the genetic cause of CGs III and I peroxisome deficiency disorders, respectively. Moreover, we found Pex5p (PTS1 receptor) to be involved in import of not only PTS1 but also PTS2 protein, using CGII ZP105 and ZP139 of complementation group III.