title: 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.
abstract: 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' end of the nd5 gene; deleted monomers deprived of cob, nd4, nd5, and the 5' 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 rotenone-resistant 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.
authors.affiliation: Genetics of Microorganisms, Department of Plant Biology, B22, University of Liège, B4000 Liège, Belgium.
authors.forename: F
authors.initials: F
authors.lastname: Duby
authors.forename: R F
authors.initials: RF
authors.lastname: Matagne
mesh_terms: D000818:Animals; D001059:Apoproteins / Q000235:genetics*; D001483:Base Sequence; D019069:Cell Respiration / Q000502:physiology; D002696:Chlamydomonas / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D003573:Cytochrome b Group / Q000235:genetics*; D045303:Cytochromes b; D004272:DNA, Mitochondrial / Q000235:genetics*; D042967:Electron Transport Complex I; D003576:Electron Transport Complex IV / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D009245:NADH Dehydrogenase / Q000235:genetics*; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics*; D010788:Photosynthesis / Q000235:genetics*; D017384:Sequence Deletion
chemical_list: D001059:Apoproteins; D003573:Cytochrome b Group; D004272:DNA, Mitochondrial; C098483:NADH dehydrogenase subunit 4; D045303:Cytochromes b; D009247:NADH, NADPH Oxidoreductases; D009245:NADH Dehydrogenase; D003576:Electron Transport Complex IV; D042967:Electron Transport Complex I