Article (Scientific journals)
A Forward Genetic Screen Identifies Mutants Deficient for Mitochondrial Complex I Assembly in Chlamydomonas Reinhardtii.
Barbieri, M. R.; Larosa, Véronique; Nouet, Cécile et al.
2011In Genetics, 188, p. 349-358
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Abstract :
[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.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Barbieri, M. R.
Larosa, Véronique ;  Université de Liège - ULiège > Département des sciences de la vie > Génétique
Nouet, Cécile  ;  Université de Liège - ULiège > Département des sciences de la vie > Génomique fonctionnelle et imagerie moléculaire végétale
Subrahmanian, N.;  Université de Liège - ULiège > Département des sciences de la vie > Génétique
Remacle, Claire  ;  Université de Liège - ULiège > Département des sciences de la vie > Génétique
Hamel, Patrice
Language :
English
Title :
A Forward Genetic Screen Identifies Mutants Deficient for Mitochondrial Complex I Assembly in Chlamydomonas Reinhardtii.
Publication date :
2011
Journal title :
Genetics
ISSN :
0016-6731
eISSN :
1943-2631
Publisher :
Genetics Society of America, United States - Maryland
Volume :
188
Pages :
349-358
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 16 May 2011

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