Protein instability and functional defects caused by mutations of dihydro-orotate dehydrogenase in Miller syndrome patients
| Autor: | Haruyoshi Yamaza, Mikako Yagi, Takeshi Uchiumi, Toshiro Saito, Kazuaki Nonaka, Dongchon Kang, Tomotake Kanki, Jing Xian Fang, Shinya Matsumoto, Shinya Takazaki, Rie Amamoto |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Ubiquinone
Mutant DHO dihydroorotate OXPHOS oxidative phosphorylation Dihydroorotate Dehydrogenase lcsh:Life lcsh:QR1-502 Mitochondrion DCPIP dichlorophenolindophenol medicine.disease_cause Biochemistry DMEM Dulbecco’s modified Eagle’s medium lcsh:Microbiology Electron Transport Complex III Missense mutation DHODH dihydro-orotate dehydrogenase mitochondrion Miller syndrome Mutation TFAM mitochondrial transcription factor A anti-HA anti-haemagglutinin DHODH-HA DHODH with a C-terminal HA tag Mitochondria Succinate Dehydrogenase protein stability UMPS UMP synthase Intermembrane space Oxidoreductases Acting on CH-CH Group Donors Micrognathism DOX doxycycline Biophysics Limb Deformities Congenital Mutation Missense Biology S2 CHX cycloheximide FBS fetal bovine serum LFN leflunomide medicine Humans Abnormalities Multiple Molecular Biology Gene Original Paper missense mutation dihydro-orotate dehydrogenase (DHODH) Cell Biology medicine.disease Molecular biology DHFR dihydrofolate reductase lcsh:QH501-531 Dihydroorotate dehydrogenase Mandibulofacial Dysostosis HeLa Cells |
| Zdroj: | Bioscience Reports, Vol 32, Iss 6 (2012) Bioscience Reports |
| ISSN: | 1573-4935 0144-8463 |
| Popis: | Miller syndrome is a recessive inherited disorder characterized by postaxial acrofacial dysostosis. It is caused by dysfunction of the DHODH (dihydroorotate dehydrogenase) gene, which encodes a key enzyme in the pyrimidine de novo biosynthesis pathway and is localized at mitochondria intermembrane space. We investigated the consequence of three missense mutations, G202A, R346W and R135C of DHODH, which were previously identified in patients with Miller syndrome. First, we established HeLa cell lines stably expressing DHODH with Miller syndrome-causative mutations: G202A, R346W and R135C. These three mutant proteins retained the proper mitochondrial localization based on immunohistochemistry and mitochondrial subfractionation studies. The G202A, R346W DHODH proteins showed reduced protein stability. On the other hand, the third one R135C, in which the mutation lies at the ubiquinone-binding site, was stable but possessed no enzymatic activity. In conclusion, the G202A and R346W mutation causes deficient protein stability, and the R135C mutation does not affect stability but impairs the substrate-induced enzymatic activity, suggesting that impairment of DHODH activity is linked to the Miller syndrome phenotype. |
| Databáze: | OpenAIRE |
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