Mitochondrial transformation and in vitro DNA delivery

in Genomics of Chloroplasts and Mitochondria (2012)

A Forward Genetic Screen Identifies Mutants Deficient for Mitochondrial Complex I Assembly in Chlamydomonas Reinhardtii.

in Genetics (2011)

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 ... [more ▼]

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. [less ▲]

Characterization of complex I mutants in Chlamydomonas reinhardtii : Role of structural subunits and identification of assembly factors.

Scientific conference (2010, January 26)

Transition Metal Nutrition: A Balance Between Deficiency and Toxicity

in Stern, David; Harris, Elizabeth H. (Eds.) The Chlamydomonas Sourcebook, volume 2: Organellar and Metabolic Processes (2009)

Eukaryotic complex I: functional diversity and experimental systems to unravel the assembly process

in Molecular Genetics & Genomics (2008), 280