Mitochondrial Genetics

Remacle, Claire; Matagne, René-Fernand

doi:10.1007/0-306-48204-5_34

Cited by 12 publications

(18 citation statements)

References 76 publications

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“…Indeed, the deletions described to date range from 0.7 to 1.7 kb and exclusively affect the left telomeric region and part or all of the cob sequence (Dorthu et al, 1992;Randolph-Anderson et al, 1993;Remacle and Matagne, 1998). In dum24 cells, two types of deleted monomers have been identified: monomers with a 3.4-kb left terminal deletion, extending from the telomere to the 3Ј end of the nd5 coding sequence, and monomers with a 5.1-kb deletion, which encompasses the telomere, cob, nd4, nd5, and the 5Ј end of the cox1 coding sequence.…”

Section: Discussionmentioning

confidence: 99%

“…The mutants lack the cytochrome pathway of the mitochondrial electron transport chain, but their respiratory activity is partially maintained via the alternative pathway of respiration. Phenotypically, the mutants have lost their capacity to grow under heterotrophic conditions (darkness and the addition of acetate as a reduced carbon source), whereas their photoautotrophic growth is barely affected (reviewed in Remacle and Matagne, 1998).…”

Section: Introductionmentioning

confidence: 99%

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Alteration of Dark Respiration and Reduction of Phototrophic Growth in a Mitochondrial DNA Deletion Mutant of Chlamydomonas Lacking cob, nd4, and the 3′ End of nd5

Duby

Matagne

1999

Plant Cell

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We describe here a new type of mitochondrial mutation ( dum24 ; for dark uniparental minus inheritance) of the unicellular photosynthetic alga Chlamydomonas reinhardtii . The mutant fails to grow under heterotrophic conditions and displays reduced growth under both photoautotrophic and mixotrophic conditions. In reciprocal crosses between mutant and wild-type cells, the meiotic progeny only inherit the phenotype of the mating-type minus parent, indicating that the dum24 mutation exclusively affects the mitochondrial genome. Digestion with various restriction enzymes followed by DNA gel blot hybridizations with specific probes demonstrated that dum24 cells contain four types of altered mitochondrial genomes: deleted monomers lacking cob , nd4 , and the 3 Ј end of the nd5 gene; deleted monomers deprived of cob , nd4 , nd5 , and the 5 Ј end of the cox1 coding sequence; and two types of dimers produced by end-to-end fusions between monomers similarly or differently deleted. Due to these mitochondrial DNA alterations, complex I activity, the cytochrome pathway of respiration, and presumably, the three phosphorylation sites associated with these enzyme activities are lacking in the mutant. The low respiratory rate of the dum24 cells results from the activities of rotenoneresistant NADH dehydrogenase, complex II, and alternative oxidase, with none of these enzymes being coupled to ATP production. To our knowledge, this type of mitochondrial mutation has never been described for photosynthetic organisms or more generally for obligate aerobes.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Alteration of Dark Respiration and Reduction of Phototrophic Growth in a Mitochondrial DNA Deletion Mutant of Chlamydomonas Lacking cob, nd4, and the 3′ End of nd5

Duby

Matagne

1999

Plant Cell

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“…The 15.8-kbp linear mitochondrial genome of C. reinhardtii encodes seven proteins involved in respiration, one protein resembling a reverse transcriptase, two ribosomal RNA genes that are fragmented and interspersed with other coding sequences, and three tRNA genes (see Remacle and Matagne, 1998). The other tRNAs have to be imported from the chloroplast or cytosol.…”

Section: Three Transformable Genetic Systemsmentioning

confidence: 99%

“…Although mitochondrial transformation has been reported for C. reinhardtii, the yield is rather low. However, the availability of several drug-resistant mitochondrial mutations offers promising new selectable markers for mitochondrial transformation (Remacle and Matagne, 1998).…”

Section: Three Transformable Genetic Systemsmentioning

confidence: 99%

Assembly, Function, and Dynamics of the Photosynthetic Machinery in Chlamydomonas reinhardtii

Rochaix

2001

Plant Physiology

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“…1). Several mutations altering mitochondrial genes encoding apocytochrome b (cob gene) and subunit 1 of cytochrome c oxidase (cox1 gene) have been characterized (11). Phenotypically, these mutants have lost the capacity to grow under heterotrophic conditions, i.e., in the dark, with acetate as carbon source.…”

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confidence: 99%

High-efficiency biolistic transformation of Chlamydomonas mitochondria can be used to insert mutations in complex I genes

Remacle

Cardol

Coosemans

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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158

131

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Mitochondrial transformation of Chlamydomonas reinhardtii has been optimized by using a particle-gun device and cloned mitochondrial DNA or PCR fragments. A respiratory-deficient strain lacking a 1.2-kb mitochondrial DNA region including the left telomere and part of the cob gene could be rescued as well as a double-frameshift mutant in the mitochondrial cox1 and nd1 genes. High transformation efficiency has been achieved (100 -250 transformants per microgram of DNA), the best results being obtained with linearized plasmid DNA. Molecular analysis of the transformants suggests that the right telomere sequence can be copied to reconstruct the left telomere by recombination. In addition, both nondeleterious and deleterious mutations could be introduced. Myxothiazol-resistant transformants have been created by introducing a nucleotide substitution into the cob gene. Similarly, an in-frame deletion of 23 codons has been created in the nd4 mitochondrial gene of both the deleted and frameshift recipient strains. These 23 codons are believed to encode the first transmembrane segment of the ND4 protein. This ⌬nd4 mutation causes a misassembly of complex I, with the accumulation of a subcomplex that is 250-kDa smaller than the wild-type complex I. The availability of efficient mitochondrial transformation in Chlamydomonas provides an invaluable tool for the study of mitochondrial biogenesis and, more specifically, for site-directed mutagenesis of mitochondrially encoded subunits of complex I, of special interest because the yeast Saccharomyces cerevisiae, whose mitochondrial genome can be manipulated virtually at will, is lacking complex I.green alga ͉ mitochondrial DNA mutagenesis ͉ telomere ͉ complex I assembly ͉ respiratory-deficient mutant

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Mitochondrial Genetics

Cited by 12 publications

References 76 publications

Alteration of Dark Respiration and Reduction of Phototrophic Growth in a Mitochondrial DNA Deletion Mutant of Chlamydomonas Lacking cob, nd4, and the 3′ End of nd5

Alteration of Dark Respiration and Reduction of Phototrophic Growth in a Mitochondrial DNA Deletion Mutant of Chlamydomonas Lacking cob, nd4, and the 3′ End of nd5

Assembly, Function, and Dynamics of the Photosynthetic Machinery in Chlamydomonas reinhardtii

High-efficiency biolistic transformation of Chlamydomonas mitochondria can be used to insert mutations in complex I genes

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