Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1977 Apr 1;73(1):56–77. doi: 10.1083/jcb.73.1.56

Nuclear mutations affecting mitochondrial structure and function in Chlamydomonas

PMCID: PMC2109904  PMID: 192732

Abstract

Wild type cells of the green alga Chlamydomonas reinhardtii can grow in the in the dark by taking up and respiring exogenously supplied acetate. Obligate photoautotrophic (dark dier, dk) mutants of this alga have been selected which grow at near wild type rates in the light, but rapidly die when transferred to darkness because of defects in mitochondrial structure and function. In crosses of the dk mutants to wild type, the majority of the mutants are inherited in a mendelian fashion, although two have been isolated which are inherited in a clearly nonmendelian fashion. Nine mendelian dk mutants have been analyzed in detail, and belong to eight different complementation groups representing eight gene loci. These mutants have been tentatively grouped into three classes on the basis of the pleiotropic nature of their phenotypic defects. Mutants in Class I have gross alterations in the ultrastructure of their mitochondrial inner membranes together with deficiencies in cytochrome oxidase and antimycin/rotenone-sensitive NADH-cytochrome c reductase activities. Mutants in Class II have a variety of less severe alterations in mitochondrial ultrastructure and deficiencies in cytochrome oxidase activity. Mutants in Class III have normal or near normal mitochondrial ultrastructure and reduced cytochrome oxidase activity. Eight of the nine mutants show corresponding reductions in cyanide-sensitive respiration.

Full Text

The Full Text of this article is available as a PDF (5.0 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ashwell M., Work T. S. The biogenesis of mitochondria. Annu Rev Biochem. 1970;39:251–290. doi: 10.1146/annurev.bi.39.070170.001343. [DOI] [PubMed] [Google Scholar]
  2. BRADY R. O. Fluoroacetyl coenzyme A. J Biol Chem. 1955 Nov;217(1):213–224. [PubMed] [Google Scholar]
  3. Bastia D., Chiang K. S., Swift H., Siersma P. Heterogeneity, complexity, and repetition of the chloroplast DNA of Chlamydomonas reinhardtii. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1157–1161. doi: 10.1073/pnas.68.6.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Borst P. Mitochondrial nucleic acids. Annu Rev Biochem. 1972;41:333–376. doi: 10.1146/annurev.bi.41.070172.002001. [DOI] [PubMed] [Google Scholar]
  5. Boynton J. E., Gillham N. W., Chabot J. F. Chloroplast ribosome deficient mutants in the green alga Chlamydomonas reinhardi and the question of chloroplast ribosome function. J Cell Sci. 1972 Mar;10(2):267–305. doi: 10.1242/jcs.10.2.267. [DOI] [PubMed] [Google Scholar]
  6. COOPERSTEIN S. J., LAZAROW A. A microspectrophotometric method for the determination of cytochrome oxidase. J Biol Chem. 1951 Apr;189(2):665–670. [PubMed] [Google Scholar]
  7. Conde M. F., Boynton J. E., Gillham N. W., Harris E. H., Tingle C. L., Wang W. L. Chloroplast genes in Chlamydomonas affecting organelle ribosomes. Genetic and biochemical analysis of analysis of antibiotic-resistant mutants at several gene loci. Mol Gen Genet. 1975 Oct 3;140(3):183–220. doi: 10.1007/BF00334266. [DOI] [PubMed] [Google Scholar]
  8. Ebersold W. T. Chlamydomonas reinhardi: heterozygous diploid strains. Science. 1967 Jul 28;157(3787):447–449. doi: 10.1126/science.157.3787.447. [DOI] [PubMed] [Google Scholar]
  9. Ebner E., Mason T. L., Schatz G. Mitochondrial assembly in respiration-deficient mutants of Saccharomyces cerevisiae. II. Effect of nuclear and extrachromosomal mutations on the formation of cytochrome c oxidase. J Biol Chem. 1973 Aug 10;248(15):5369–5378. [PubMed] [Google Scholar]
  10. Ebner E., Mennucci L., Schatz G. Mitochondrial assembly in respiration-deficient mutants of Saccharomyces cerevisiae. I. Effect of nuclear mutations on mitochondrial protein synthesis. J Biol Chem. 1973 Aug 10;248(15):5360–5368. [PubMed] [Google Scholar]
  11. Ebner E., Schatz G. Mitochondrial assembly in respiration-deficient mutants of Saccharomyces cerevisiae. 3. A nuclear mutant lacking mitochondrial adenosine triphosphatase. J Biol Chem. 1973 Aug 10;248(15):5379–5384. [PubMed] [Google Scholar]
  12. Edwards D. L., Kwiecinski F. Altered mitochondrial respiration in a chromosomal mutant of Neurospora crassa. J Bacteriol. 1973 Nov;116(2):610–618. doi: 10.1128/jb.116.2.610-618.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Edwards D. L., Rosenberg E. Regulation of cyanide-insensitive respiration in Neurospora. Eur J Biochem. 1976 Feb 16;62(2):217–221. doi: 10.1111/j.1432-1033.1976.tb10150.x. [DOI] [PubMed] [Google Scholar]
  14. GILLHAM N. W. TRANSMISSION AND SEGREGATION OF A NON-CHROMOSOMAL FACTOR CONTROLLING STREPTOMYCIN RESISTANCE IN DIPLOID CHLAMYDOMONAS. Nature. 1963 Oct 19;200:294–294. doi: 10.1038/200294a0. [DOI] [PubMed] [Google Scholar]
  15. Henry M. F., Nyns E. D. Cyanide-insensitive respiration. An alternative mitochondrial pathway. Subcell Biochem. 1975 Mar;4(1):1–65. [PubMed] [Google Scholar]
  16. Kolarov J., Subík J., Kovac L. Oxidative phosphorylation in yeast. IX. Modification of the mitochondrial adenine nucleotide translocation system in the oxidative phosphorylation-deficient mutant op 1 . Biochim Biophys Acta. 1972 Jun 23;267(3):465–478. doi: 10.1016/0005-2728(72)90174-0. [DOI] [PubMed] [Google Scholar]
  17. LEVINE R. P., EBERSOLD W. T. The genetics and cytology of Chlamydomonas. Annu Rev Microbiol. 1960;14:197–216. doi: 10.1146/annurev.mi.14.100160.001213. [DOI] [PubMed] [Google Scholar]
  18. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  19. Lee R. W., Jones R. F. Induction of Mendelian and non-Mendelian streptomycin resistant mutants during the synchronous cell cycle of Chlamydomonas reinhardtii. Mol Gen Genet. 1973 Mar 1;121(2):99–108. doi: 10.1007/BF00277524. [DOI] [PubMed] [Google Scholar]
  20. MORRISON J. F., PETERS R. A. Biochemistry of fluoroacetate poisoning: the effect of fluorocitrate on purified aconitase. Biochem J. 1954 Nov;58(3):473–479. doi: 10.1042/bj0580473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. MUNKRES K. D., RICHARDS F. M. GENETIC ALTERATION OF NEUROSPORA MALATE DEHYDROGENASE. Arch Biochem Biophys. 1965 Mar;109:457–465. doi: 10.1016/0003-9861(65)90390-5. [DOI] [PubMed] [Google Scholar]
  22. Müller M., Hogg J. F., De Duve C. Distribution of tricarboxylic acid cycle enzymes and glyoxylate cycle enzymes between mitochondria and peroxisomes in Tetrahymena pyriformis. J Biol Chem. 1968 Oct 25;243(20):5385–5395. [PubMed] [Google Scholar]
  23. Ogur M., Coker L., Ogur S. Glutamate auxotrophs in Saccharomyces 1. I. The biochemical lesion in the glt-1 mutants-2. Biochem Biophys Res Commun. 1964;14:193–197. doi: 10.1016/0006-291x(64)90254-2. [DOI] [PubMed] [Google Scholar]
  24. Richter D. Production of mitochondrial peptide-chain elongation factors in yeast deficient in mitochondrial deoxyribonucleic acid. Biochemistry. 1971 Nov 23;10(24):4422–4425. doi: 10.1021/bi00800a011. [DOI] [PubMed] [Google Scholar]
  25. SMITH L. Spectrophotometric assay of cytochrome c oxidase. Methods Biochem Anal. 1955;2:427–434. doi: 10.1002/9780470110188.ch13. [DOI] [PubMed] [Google Scholar]
  26. Santarius K. A., Heber U. Changes in the intracellular levels of ATP, ADP, AMP and P1 and regulatory function of the adenylate system in leaf cells during photosynthesis. Biochim Biophys Acta. 1965 May 25;102(1):39–54. doi: 10.1016/0926-6585(65)90201-3. [DOI] [PubMed] [Google Scholar]
  27. Seligman A. M., Karnovsky M. J., Wasserkrug H. L., Hanker J. S. Nondroplet ultrastructural demonstration of cytochrome oxidase activity with a polymerizing osmiophilic reagent, diaminobenzidine (DAB). J Cell Biol. 1968 Jul;38(1):1–14. doi: 10.1083/jcb.38.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sherman F., Stewart J. W., Jackson M., Gilmore R. A., Parker J. H. Mutants of yeast defective in iso-1-cytochrome c. Genetics. 1974 Jun;77(2):255–284. doi: 10.1093/genetics/77.2.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sherman F., Stewart J. W., Parker J. H., Inhaber E., Shipman N. A., Putterman G. J., Gardisky R. L., Margoliash E. The mutational alteration of the primary structure of yeast iso-1-cytochrome c. J Biol Chem. 1968 Oct 25;243(20):5446–5456. [PubMed] [Google Scholar]
  30. Somlo M. Respiration-linked ATP formation by an "oxidative phosphorylation mutant" of yeast. Arch Biochem Biophys. 1970 Jan;136(1):122–133. doi: 10.1016/0003-9861(70)90334-6. [DOI] [PubMed] [Google Scholar]
  31. Sueoka N. MITOTIC REPLICATION OF DEOXYRIBONUCLEIC ACID IN CHLAMYDOMONAS REINHARDI. Proc Natl Acad Sci U S A. 1960 Jan;46(1):83–91. doi: 10.1073/pnas.46.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tzagoloff A., Akai A., Needleman R. B. Assembly of the mitochondrial membrane system. Characterization of nuclear mutants of Saccharomyces cerevisiae with defects in mitochondrial ATPase and respiratory enzymes. J Biol Chem. 1975 Oct 25;250(20):8228–8235. [PubMed] [Google Scholar]
  33. Wang W. Y., Wang W. L., Boynton J. E., Gillham N. W. Genetic control of chlorophyll biosynthesis in Chlamydomonas. Analysis of mutants at two loci mediating the conversion of protoporphyrin-IX to magnesium protoporphyrin. J Cell Biol. 1974 Dec;63(3):806–823. doi: 10.1083/jcb.63.3.806. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wells R., Sager R. Denaturation and the renaturation kinetics of chloroplast DNA from Chlamydomonas reinhardi. J Mol Biol. 1971 Jun 14;58(2):611–622. doi: 10.1016/0022-2836(71)90375-5. [DOI] [PubMed] [Google Scholar]
  35. Wiseman A., Gillham N. W., Boynton J. E. The mitochondrial genome of Chlamydomonas. II. Genetic analysis of non-mendelian obligate photautotrophic mutants. Mol Gen Genet. 1977 Jan 18;150(2):109–118. doi: 10.1007/BF00695390. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES