Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1987 Mar;84(6):1555–1559. doi: 10.1073/pnas.84.6.1555

Inhibition of glycolate and D-lactate metabolism in a Chlamydomonas reinhardtii mutant deficient in mitochondrial respiration

Diane W Husic 1,*, N E Tolbert 1,†
PMCID: PMC304474  PMID: 16578800

Abstract

The possibility that glycolate oxidation in unicellular green algae is linked to mitochondrial electron transport, rather than to peroxisomal metabolism as in higher plants and animals, was studied in a mutant of Chlamydomonas reinhardtii (dk97) deficient in cytochrome oxidase. This mutant had normal rates of dark respiration (40 ± 15 μmol of O2 uptake per hr per mg of chlorophyll) but had only 11% of wild-type levels of cytochrome oxidase activity. Salicylhydroxamic acid (SHAM) reduced the dark respiration rate of dk97 cells by 71%, but cyanide did not significantly inhibit this rate. During photosynthesis in the presence of SHAM, glycolate oxidation was blocked, resulting in glycolate accumulation and excretion by mutant cells but not by wild-type Chlamydomonas. D-Lactate, which accumulated after brief periods of anaerobiosis in Chlamydomonas, was reoxidized by air-grown cells only aerobically in the light, and reoxidation of D-lactate was blocked by SHAM in the dk97 cells. Thus, glycolate and D-lactate dehydrogenase activities are both linked to mitochondrial electron transport in Chlamydomonas. During photosynthetic 14CO2 fixation by dk97 cells in the presence of SHAM, 14C-labeled tricarboxylic acid cycle intermediates accumulated, indicating that, in Chlamydomonas, mitochondrial respiration functions during photosynthesis.

Keywords: CN-insensitive respiration, cytochrome c mutant, glycolate and D-lactate dehydrogenase, peroxisomes

Full text

PDF
1555

Selected References

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

  1. Beezley B. B., Gruber P. J., Frederick S. E. Cytochemical localization of glycolate dehydrogenase in mitochondria of chlamydomonas. Plant Physiol. 1976 Sep;58(3):315–319. doi: 10.1104/pp.58.3.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Collins N., Brown R. H., Merrett M. J. Oxidative phosphorylation during glycollate metabolism in mitochondria from phototrophic Euglena gracilis. Biochem J. 1975 Sep;150(3):373–377. doi: 10.1042/bj1500373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Everse J., Kaplan N. O. Lactate dehydrogenases: structure and function. Adv Enzymol Relat Areas Mol Biol. 1973;37:61–133. doi: 10.1002/9780470122822.ch2. [DOI] [PubMed] [Google Scholar]
  4. GREGOLIN C., SINGER T. P. The lactic dehydrogenase of yeast. III. D(-)Lactic cytochrome c reductase, a zinc-flavoprotein from aerobic yeast. Biochim Biophys Acta. 1963 Feb 12;67:201–218. doi: 10.1016/0006-3002(63)91818-3. [DOI] [PubMed] [Google Scholar]
  5. Grodzinski B., Colman B. Intracellular localization of glycolate dehydrogenase in a blue-green alga. Plant Physiol. 1976 Aug;58(2):199–202. doi: 10.1104/pp.58.2.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Husic D. W., Tolbert N. E. Anaerobic Formation of d-Lactate and Partial Purification and Characterization of a Pyruvate Reductase from Chlamydomonas reinhardtii. Plant Physiol. 1985 Jun;78(2):277–284. doi: 10.1104/pp.78.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ingledew W. J., Poole R. K. The respiratory chains of Escherichia coli. Microbiol Rev. 1984 Sep;48(3):222–271. doi: 10.1128/mr.48.3.222-271.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kaback H. R. Transport studies in bacterial membrane vesicles. Science. 1974 Dec 6;186(4167):882–892. doi: 10.1126/science.186.4167.882. [DOI] [PubMed] [Google Scholar]
  9. Moroney J. V., Husic H. D., Tolbert N. E. Effect of Carbonic Anhydrase Inhibitors on Inorganic Carbon Accumulation by Chlamydomonas reinhardtii. Plant Physiol. 1985 Sep;79(1):177–183. doi: 10.1104/pp.79.1.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Moroney J. V., Wilson B. J., Tolbert N. E. Glycolate Metabolism and Excretion by Chlamydomonas reinhardtii. Plant Physiol. 1986 Nov;82(3):821–826. doi: 10.1104/pp.82.3.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Nelson E. B., Tolbert N. E. Glycolate dehydrogenase in green algae. Arch Biochem Biophys. 1970 Nov;141(1):102–110. doi: 10.1016/0003-9861(70)90112-8. [DOI] [PubMed] [Google Scholar]
  12. Peltier G., Thibault P. Light-Dependent Oxygen Uptake, Glycolate, and Ammonia Release in l-Methionine Sulfoximine-Treated Chlamydomonas. Plant Physiol. 1985 Feb;77(2):281–284. doi: 10.1104/pp.77.2.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Peltier G., Thibault P. O(2) uptake in the light in chlamydomonas: evidence for persistent mitochondrial respiration. Plant Physiol. 1985 Sep;79(1):225–230. doi: 10.1104/pp.79.1.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Poerio E., Davies D. D. A comparison of potato and vertebrate lactate dehydrogenases. Biochem J. 1980 Nov 1;191(2):341–348. doi: 10.1042/bj1910341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. SYRETT P. J., WONG H. A. THE FERMENTATION OF GLUCOSE BY CHLORELLA VULGARIS. Biochem J. 1963 Nov;89:308–315. doi: 10.1042/bj0890308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sandmann G., Malkin R. Light inhibition of respiration is due to a dual function of the cytochrome b6-f complex and the plastocyanin/cytochrome c-553 pool in Aphanocapsa. Arch Biochem Biophys. 1984 Oct;234(1):105–111. doi: 10.1016/0003-9861(84)90329-1. [DOI] [PubMed] [Google Scholar]
  17. Schonbaum G. R., Bonner W. D., Jr, Storey B. T., Bahr J. T. Specific inhibition of the cyanide-insensitive respiratory pathway in plant mitochondria by hydroxamic acids. Plant Physiol. 1971 Jan;47(1):124–128. doi: 10.1104/pp.47.1.124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Tolbert N. E., Harrison M., Selph N. Aminooxyacetate stimulation of glycolate formation and excretion by chlamydomonas. Plant Physiol. 1983 Aug;72(4):1075–1083. doi: 10.1104/pp.72.4.1075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Walsh C. T., Schonbrunn A., Lockridge O., Massey V., Abeles R. H. Inactivation of a flavoprotein, lactate oxidase, by an acetylenic substrate. J Biol Chem. 1972 Sep 25;247(18):6004–6006. [PubMed] [Google Scholar]
  21. Wiseman A., Gillham N. W., Boynton J. E. Nuclear mutations affecting mitochondrial structure and function in Chlamydomonas. J Cell Biol. 1977 Apr;73(1):56–77. doi: 10.1083/jcb.73.1.56. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES