期刊
GENETICS
卷 188, 期 2, 页码 349-U195出版社
GENETICS SOCIETY AMERICA
DOI: 10.1534/genetics.111.128827
关键词
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资金
- United Mitochondrial Disease Foundation
- F.R.S.-FNRS [1.5.255.08, 2.4638.05, 2.4601.08]
- Action de la Recherche Concertee [ARC07/12-04]
- Fonds de la Recherche Scientifique - Fonds National de la Recherche Scientifique, Formation a la Recherche dans l'Industrie et l'Agriculture fellowship
- Action Centre National de la Recherche Scientifique (C.N.R.S)-US
- [GA 245070]
Mitochondrial complex I is the largest multimeric enzyme of the respiratory chain. The lack of a model system with facile genetics has limited the molecular dissection of complex I assembly. Using Chlamydomonas reinhardtii as an experimental system to screen for complex I defects, we isolated, via forward genetics, amc1-7 nuclear mutants (for assembly of mitochondrial complex I) displaying reduced or no complex I activity. Blue native (BN)-PAGE and immunoblot analyses revealed that amc3 and amc4 accumulate reduced levels of the complex I holoenzyme (950 kDa) while all other amc mutants fail to accumulate a mature complex. In amc1, -2, -5-7, the detection of a 700 kDa subcomplex retaining NADH dehydrogenase activity indicates an arrest in the assembly process. Genetic analyses established that amc5 and amc7 are alleles of the same locus while amc1-4 and amc6 define distinct complementation groups. The locus defined by the amc5 and amc7 alleles corresponds to the NUOB10 gene, encoding PDSW, a subunit of the membrane arm of complex I. This is the first report of a forward genetic screen yielding the isolation of complex I mutants. This work illustrates the potential of using Chlamydomonas as a genetically tractable organism to decipher complex I manufacture.
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