Abstract
The sequence and organization of the Chlamydomonas reinhardtii genes encoding cytochrome c 1 ( Cyc1) and the Rieske-type iron-sulfur protein ( Isp), two key nucleus-encoded subunits of the mitochondrial cytochrome bc 1 complex, are presented. Southern hybridization analysis indicates that both Cyc1 and Isp are present as single-copy genes in C. reinhardtii. The Cyc1 gene spans 6404 bp and contains six introns, ranging from 178 to 1134 bp in size. The Isp gene spans 1238 bp and contains four smaller introns, ranging in length from 83 to 167 bp. In both genes, the intron/exon junctions follow the GT/AG rule. Internal conserved sequences were identified in only some of the introns in the Cyc1 gene. The levels of expression of Isp and Cyc1 genes are comparable in wild-type C. reinhardtii cells and in a mutant strain carrying a deletion in the mitochondrial gene for cytochrome b (dum-1). Nevertheless, no accumulation of the nucleus-encoded cytochrome c 1 or of core proteins I and II was observed in the membranes of the respiratory mutant. These data show that, in the green alga C. reinhardtii, the subunits of the cytochrome bc 1 complex fail to assemble properly in the absence of cytochrome b.
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Acknowledgments
The biochemical characterization of the dum-1 mutant was initiated in the Laboratory of Dr. J.-L Popot (IBPC, Paris). The authors wish to thank Drs. E. H. Harris (Duke University) and C. D. Silflow (University of Minnesota), who kindly provided us with DNA probes used in this work; Dr. H.-P. Braun (Institut fur Angewandte Genetik, Universität Hannover) and Dr. G. Schatz (Biozentrum der Universität Basel) for the gift of the antisera; Drs. J. Girard-Bascou (IBPC, Paris), E. H. Harris (Duke University) and R.F. Matagne (Laboratoire de Génétique des Microorganismes, Liège) for the gift of several probes and strains. We express our gratitude to T. Ballado (IFC, UNAM) for her valuable advice along the work, and to Dr. G. Dreyfus (IFC, UNAM) for the use of some of his laboratory equipment and chemicals. The technical skills of Guadalupe Códiz (Unidad de Biología Molecular, IFC, UNAM) in expert nucleotide sequencing are also acknowledged. This work was supported by grants from CONACyT, Mexico (27754 N); and from DGAPA, UNAM, Mexico (IN207201). RvL received a fellowship from DGEP (UNAM). LGF was supported by a grant from Carl Trygger's Foundation for Scientific Research
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Atteia, A., van Lis, R., Wetterskog, D. et al. Structure, organization and expression of the genes encoding mitochondrial cytochrome c 1 and the Rieske iron-sulfur protein in Chlamydomonas reinhardtii . Mol Gen Genomics 268, 637–644 (2003). https://doi.org/10.1007/s00438-002-0779-x
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DOI: https://doi.org/10.1007/s00438-002-0779-x