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
. 1983 Oct;80(20):6293–6297. doi: 10.1073/pnas.80.20.6293

Rapid recovery of chloroplast mutations affecting ribulosebisphosphate carboxylase/oxygenase in Chlamydomonas reinhardtii

Robert J Spreitzer 1,2,*, William L Ogren 1,2
PMCID: PMC394283  PMID: 16593382

Abstract

Based on the unique ability of chloroplast genes to recombine in Chlamydomonas reinhardtii, a collection of acetate-requiring mutants was screened for recombination with a mutation affecting ribulose-1,5-bisphosphate carboxylase/oxygenase [Rbu-1,5-P2 carboxylase/oxygenase; 3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39]. This chloroplast mutation, rcl-u-1-10-6C, causes the absence of Rbu-1,5-P2 carboxylase/oxygenase activities and alters the isoelectric point of the larger subunit. Several mutants that displayed little or no recombination with 10-6C were recovered, and two lacked carboxylase activity. These new chloroplast mutants lack both large and small Rbu-1,5-P2 carboxylase/oxygenase subunits. The approach demonstrated here permits the routine recovery of chloroplast mutations affecting this enzyme. Multiple mutations in the Rbu-1,5-P2 carboxylase/oxygenase large-subunit gene can be used to investigate the function and regulation of this enzyme and the regulation of chloroplast genes in general.

Keywords: photosynthesis, photorespiration, plant productivity, gene regulation, chloroplast loci

Full text

PDF
6293

Images in this article

6295Image
on p.6295
6295Image
on p.6295
6296Image
on p.6296

Selected References

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

  1. Baker T. S., Eisenberg D., Eiserling F. A., Weissman L. The structure of form I crystals of D-ribulose-1,5-diphosphate carboxylase. J Mol Biol. 1975 Feb 5;91(4):391–399. doi: 10.1016/0022-2836(75)90267-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barraclough R., Ellis R. J. The biosynthesis of ribulose bisphosphate carboxylase. Uncoupling of the synthesis of the large and small subunits in isolated soybean leaf cells. Eur J Biochem. 1979 Feb 15;94(1):165–177. doi: 10.1111/j.1432-1033.1979.tb12883.x. [DOI] [PubMed] [Google Scholar]
  3. Bedbrook J. R., Coen D. M., Beaton A. R., Bogorad L., Rich A. Location of the single gene for the large subunit of ribulosebisphosphate carboxylase on the maize chloroplast chromosome. J Biol Chem. 1979 Feb 10;254(3):905–910. [PubMed] [Google Scholar]
  4. Bedbrook J. R., Link G., Coen D. M., Bogorad L. Maize plastid gene expressed during photoregulated development. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3060–3064. doi: 10.1073/pnas.75.7.3060. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Cashmore A. R. Reiteration frequency of the gene coding for the small subunit of ribulose--1,5--bisphosphate carboxylase. Cell. 1979 Jun;17(2):383–388. doi: 10.1016/0092-8674(79)90164-8. [DOI] [PubMed] [Google Scholar]
  7. Chua N. H., Bennoun P. Thylakoid membrane polypeptides of Chlamydomonas reinhardtii: wild-type and mutant strains deficient in photosystem II reaction center. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2175–2179. doi: 10.1073/pnas.72.6.2175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chua N. H., Schmidt G. W. Post-translational transport into intact chloroplasts of a precursor to the small subunit of ribulose-1,5-bisphosphate carboxylase. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6110–6114. doi: 10.1073/pnas.75.12.6110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Coen D. M., Bedbrook J. R., Bogorad L., Rich A. Maize chloroplast DNA fragment encoding the large subunit of ribulosebisphosphate carboxylase. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5487–5491. doi: 10.1073/pnas.74.12.5487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dron M., Rahire M., Rochaix J. D., Mets L. First DNA sequence of a chloroplast mutation: a missense alteration in the ribulosebisphosphate carboxylase large subunit gene. Plasmid. 1983 May;9(3):321–324. doi: 10.1016/0147-619x(83)90009-4. [DOI] [PubMed] [Google Scholar]
  11. Feierabend J., Wildner G. Formation of the small subunit in the absence of the large subunit of ribulose 1,5-bisphosphate carboxylase in 70 S ribosome-deficient rye leaves. Arch Biochem Biophys. 1978 Mar;186(2):283–291. doi: 10.1016/0003-9861(78)90437-x. [DOI] [PubMed] [Google Scholar]
  12. Givan A. L., Criddle R. S. Ribulosediphosphate carboxylase from Chlamydomonas reinhardi: purification, properties and its mode of synthesis in the cell. Arch Biochem Biophys. 1972 Mar;149(1):153–163. doi: 10.1016/0003-9861(72)90309-8. [DOI] [PubMed] [Google Scholar]
  13. Grant D. M., Gillham N. W., Boynton J. E. Inheritance of chloroplast DNA in Chlamydomonas reinhardtii. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6067–6071. doi: 10.1073/pnas.77.10.6067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gray J. C., Kerwick R. G. An immunological investigation of the structure and function of ribulose 1,5-bisphosphate carboxylase. Eur J Biochem. 1974 May 15;44(2):481–489. doi: 10.1111/j.1432-1033.1974.tb03506.x. [DOI] [PubMed] [Google Scholar]
  15. Harris E. H., Boynton J. E., Gillham N. W. Chloroplast ribosome biogenesis in Chlamydomonas. Selection and characterization of mutants blocked in ribosome formation. J Cell Biol. 1974 Oct;63(1):160–179. doi: 10.1083/jcb.63.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Iwanij V., Chua N. H., Siekevitz P. Synthesis and turnover of ribulose biphosphate carboxylase and of its subunits during the cell cycle of Chlamydomonas reinhardtii. J Cell Biol. 1975 Mar;64(3):572–585. doi: 10.1083/jcb.64.3.572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Laing W. A. Regulation of Soybean Net Photosynthetic CO(2) Fixation by the Interaction of CO(2), O(2), and Ribulose 1,5-Diphosphate Carboxylase. Plant Physiol. 1974 Nov;54(5):678–685. doi: 10.1104/pp.54.5.678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Leto K. J., Keresztes A., Arntzen C. J. Nuclear Involvement in the Appearance of a Chloroplast-Encoded 32,000 Dalton Thylakoid Membrane Polypeptide Integral to the Photosystem II Complex. Plant Physiol. 1982 Jun;69(6):1450–1458. doi: 10.1104/pp.69.6.1450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Link G., Coen D. M., Bogorad L. Differential expression of the gene for the large subunit of ribulose bisphosphate carboxylase in maize leaf cell types. Cell. 1978 Nov;15(3):725–731. doi: 10.1016/0092-8674(78)90258-1. [DOI] [PubMed] [Google Scholar]
  20. Mattoo A. K., Pick U., Hoffman-Falk H., Edelman M. The rapidly metabolized 32,000-dalton polypeptide of the chloroplast is the "proteinaceous shield" regulating photosystem II electron transport and mediating diuron herbicide sensitivity. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1572–1576. doi: 10.1073/pnas.78.3.1572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Norton I. L., Welch M. H., Hartman F. C. Evidence for essential lysyl residues in ribulosebisphosphate carboxylase by use of the affinity label 3-bromo-1,4-dihydroxy-2-butanone 1,4-bisphosphate. J Biol Chem. 1975 Oct 25;250(20):8062–8068. [PubMed] [Google Scholar]
  22. Pfister K., Steinback K. E., Gardner G., Arntzen C. J. Photoaffinity labeling of an herbicide receptor protein in chloroplast membranes. Proc Natl Acad Sci U S A. 1981 Feb;78(2):981–985. doi: 10.1073/pnas.78.2.981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shepherd H. S., Boynton J. E., Gillham N. W. Mutations in nine chloroplast loci of Chlamydomonas affecting different photosynthetic functions. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1353–1357. doi: 10.1073/pnas.76.3.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Smith S. M., Ellis R. J. Light-stimulated accumulation of transcripts of nuclear and chloroplast genes for ribulosebisphosphate carboxylase. J Mol Appl Genet. 1981;1(2):127–137. [PubMed] [Google Scholar]
  25. Spreitzer R. J., Mets L. Photosynthesis-deficient Mutants of Chlamydomonas reinhardii with Associated Light-sensitive Phenotypes. Plant Physiol. 1981 Mar;67(3):565–569. doi: 10.1104/pp.67.3.565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Spreitzer R. J., Ogren W. L. Nuclear Suppressors of the Photosensitivity Associated with Defective Photosynthesis in Chlamydomonas reinhardii. Plant Physiol. 1983 Jan;71(1):35–39. doi: 10.1104/pp.71.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wurtz E. A., Boynton J. E., Gillham N. W. Perturbation of chloroplast DNA amounts and chloroplast gene transmission in Chlamydomonas reinhardtii by 5-fluorodeoxyuridine. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4552–4556. doi: 10.1073/pnas.74.10.4552. [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