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. 1996 Oct 15;93(21):12031–12034. doi: 10.1073/pnas.93.21.12031

Discovery of an algal mitochondrial carbonic anhydrase: molecular cloning and characterization of a low-CO2-induced polypeptide in Chlamydomonas reinhardtii.

M Eriksson 1, J Karlsson 1, Z Ramazanov 1, P Gardeström 1, G Samuelsson 1
PMCID: PMC1074525  PMID: 8876257

Abstract

In green unicellular algae, several polypeptides are induced upon exposure to limiting CO2. We report here on the localization and characterization of one of these, a 22-kDa polypeptide in Chlamydomonas reinhardtii. This nuclear-encoded polypeptide is induced in the mitochondria by a lowering of the partial pressure of CO2 in the growth medium from 5% to air CO2 levels. Sequencing of two different cDNA clones coding for the polypeptide identified it as a 20.7-kDa beta-type carbonic anhydrase (CA; carbonate dehydratase, carbonate hydro-lyase, EC 4.2.1.1). The two clones differ in their nucleotide sequences but code for identical proteins, showing that this CA is encoded by at least two genes. Northern blot hybridization reveals that mRNA transcripts are only present in cells transferred to air CO2 levels. A comparison of the deduced amino acid sequence with those of other beta-CAs shows the largest degree of similarity with CA from the cyanobacterium Synechocystis (50% identity and 66% similarity). To our knowledge, this is the first identification and characterization of a mitochondrial CA from a photosynthetic organism.

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Selected References

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

  1. Bailly J., Coleman J. R. Effect of CO(2) Concentration on Protein Biosynthesis and Carbonic Anhydrase Expression in Chlamydomonas reinhardtii. Plant Physiol. 1988 Aug;87(4):833–840. doi: 10.1104/pp.87.4.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bracey M. H., Christiansen J., Tovar P., Cramer S. P., Bartlett S. G. Spinach carbonic anhydrase: investigation of the zinc-binding ligands by site-directed mutagenesis, elemental analysis, and EXAFS. Biochemistry. 1994 Nov 8;33(44):13126–13131. doi: 10.1021/bi00248a023. [DOI] [PubMed] [Google Scholar]
  3. Broun G., Selegny E., Minh C. T., Thomas D. Facilitated transport of CO(2) across a membrane bearing carbonic anhydrase. FEBS Lett. 1970 Apr 16;7(3):223–226. doi: 10.1016/0014-5793(70)80166-1. [DOI] [PubMed] [Google Scholar]
  4. Burow M. D., Chen Z. Y., Mouton T. M., Moroney J. V. Isolation of cDNA clones of genes induced upon transfer of Chlamydomonas reinhardtii cells to low CO2. Plant Mol Biol. 1996 May;31(2):443–448. doi: 10.1007/BF00021807. [DOI] [PubMed] [Google Scholar]
  5. Coleman J. R., Grossman A. R. Biosynthesis of carbonic anhydrase in Chlamydomonas reinhardtii during adaptation to low CO(2). Proc Natl Acad Sci U S A. 1984 Oct;81(19):6049–6053. doi: 10.1073/pnas.81.19.6049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dodgson S. J., Forster R. E., 2nd Carbonic anhydrase: inhibition results in decreased urea production by hepatocytes. J Appl Physiol (1985) 1986 Feb;60(2):646–652. doi: 10.1152/jappl.1986.60.2.646. [DOI] [PubMed] [Google Scholar]
  7. Dodgson S. J., Forster R. E., 2nd Inhibition of CA V decreases glucose synthesis from pyruvate. Arch Biochem Biophys. 1986 Nov 15;251(1):198–204. doi: 10.1016/0003-9861(86)90066-4. [DOI] [PubMed] [Google Scholar]
  8. Enns T. Facilitation by carbonic anhydrase of carbon dioxide transport. Science. 1967 Jan 6;155(3758):44–47. doi: 10.1126/science.155.3758.44. [DOI] [PubMed] [Google Scholar]
  9. Eriksson M., Gardestrom P., Samuelsson G. Isolation, Purification, and Characterization of Mitochondria from Chlamydomonas reinhardtii. Plant Physiol. 1995 Feb;107(2):479–483. doi: 10.1104/pp.107.2.479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fukuzawa H., Fujiwara S., Tachiki A., Miyachi S. Nucleotide sequences of two genes CAH1 and CAH2 which encode carbonic anhydrase polypeptides in Chlamydomonas reinhardtii. Nucleic Acids Res. 1990 Nov 11;18(21):6441–6442. doi: 10.1093/nar/18.21.6441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fukuzawa H., Fujiwara S., Yamamoto Y., Dionisio-Sese M. L., Miyachi S. cDNA cloning, sequence, and expression of carbonic anhydrase in Chlamydomonas reinhardtii: regulation by environmental CO2 concentration. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4383–4387. doi: 10.1073/pnas.87.11.4383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hewett-Emmett D., Tashian R. E. Functional diversity, conservation, and convergence in the evolution of the alpha-, beta-, and gamma-carbonic anhydrase gene families. Mol Phylogenet Evol. 1996 Feb;5(1):50–77. doi: 10.1006/mpev.1996.0006. [DOI] [PubMed] [Google Scholar]
  13. Hiltonen T., Karlsson J., Palmqvist K., Clarke A. K., Samuelsson G. Purification and characterisation of an intracellular carbonic anhydrase from the unicellular green alga Coccomyxa. Planta. 1995;195(3):345–351. doi: 10.1007/BF00202591. [DOI] [PubMed] [Google Scholar]
  14. Husic H. D., Marcus C. A. Identification of Intracellular Carbonic Anhydrase in Chlamydomonas reinhardtii with a Carbonic Anhydrase-Directed Photoaffinity Label. Plant Physiol. 1994 May;105(1):133–139. doi: 10.1104/pp.105.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Johanningmeier U., Howell S. H. Regulation of light-harvesting chlorophyll-binding protein mRNA accumulation in Chlamydomonas reinhardi. Possible involvement of chlorophyll synthesis precursors. J Biol Chem. 1984 Nov 10;259(21):13541–13549. [PubMed] [Google Scholar]
  16. Kaneko T., Tanaka A., Sato S., Kotani H., Sazuka T., Miyajima N., Sugiura M., Tabata S. Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. I. Sequence features in the 1 Mb region from map positions 64% to 92% of the genome. DNA Res. 1995 Aug 31;2(4):153-66, 191-8. doi: 10.1093/dnares/2.4.153. [DOI] [PubMed] [Google Scholar]
  17. Karlsson J., Hiltonen T., Husic H. D., Ramazanov Z., Samuelsson G. Intracellular carbonic anhydrase of Chlamydomonas reinhardtii. Plant Physiol. 1995 Oct;109(2):533–539. doi: 10.1104/pp.109.2.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Manuel L. J., Moroney J. V. Inorganic Carbon Accumulation by Chlamydomonas reinhardtii: New Proteins are made During Adaptation to Low CO(2). Plant Physiol. 1988 Oct;88(2):491–496. doi: 10.1104/pp.88.2.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marek L. F., Spalding M. H. Changes in Photorespiratory Enzyme Activity in Response to Limiting CO(2) in Chlamydomonas reinhardtii. Plant Physiol. 1991 Sep;97(1):420–425. doi: 10.1104/pp.97.1.420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Oliver D. J., Raman R. Glycine decarboxylase: protein chemistry and molecular biology of the major protein in leaf mitochondria. J Bioenerg Biomembr. 1995 Aug;27(4):407–414. doi: 10.1007/BF02110003. [DOI] [PubMed] [Google Scholar]
  23. Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
  24. Ramazanov Z., Mason C. B., Geraghty A. M., Spalding M. H., Moroney J. V. The Low CO2-Inducible 36-Kilodalton Protein Is Localized to the Chloroplast Envelope of Chlamydomonas reinhardtii. Plant Physiol. 1993 Apr;101(4):1195–1199. doi: 10.1104/pp.101.4.1195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Spalding M. H., Jeffrey M. Membrane-Associated Polypeptides Induced in Chlamydomonas by Limiting CO(2) Concentrations. Plant Physiol. 1989 Jan;89(1):133–137. doi: 10.1104/pp.89.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Tashian R. E. Genetics of the mammalian carbonic anhydrases. Adv Genet. 1992;30:321–356. doi: 10.1016/s0065-2660(08)60323-5. [DOI] [PubMed] [Google Scholar]

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