Reference : A Forward Genetic Screen Identifies Mutants Deficient for Mitochondrial Complex I Assemb...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/91120
A Forward Genetic Screen Identifies Mutants Deficient for Mitochondrial Complex I Assembly in Chlamydomonas Reinhardtii.
English
Barbieri, M. R. [> > > >]
Larosa, Véronique mailto [Université de Liège - ULg > Département des sciences de la vie > Génétique >]
Nouet, Cécile mailto [Université de Liège - ULg > Département des sciences de la vie > Génomique fonctionnelle et imagerie moléculaire végétale >]
Subrahmanian, N. [Université de Liège - ULg > Département des sciences de la vie > Génétique]
Remacle, Claire mailto [Université de Liège - ULg > Département des sciences de la vie > Génétique >]
Hamel, Patrice mailto [> >]
2011
Genetics
188
349-358
Yes (verified by ORBi)
International
0016-6731
1943-2631
[en] 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 to 7 nuclear mutants (for assembly of mitochondrial complex I) displaying reduced or no Complex I activity. 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, 6, 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 to 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.
http://hdl.handle.net/2268/91120
10.1534/genetics.111.128827

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