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. 1994 Jun 15;13(12):2747–2754. doi: 10.1002/j.1460-2075.1994.tb06568.x

The chloroplast ycf8 open reading frame encodes a photosystem II polypeptide which maintains photosynthetic activity under adverse growth conditions.

C Monod 1, Y Takahashi 1, M Goldschmidt-Clermont 1, J D Rochaix 1
PMCID: PMC395154  PMID: 8026459

Abstract

We have engineered and analyzed a chloroplast mutant of Chlamydomonas reinhardtii that lacks ycf8, the chloroplast open reading frame 8, which is highly conserved in location and predicted amino acid sequence in land plants and C.reinhardtii. The ycf8 sequence was replaced with the aadA cassette which confers resistance to spectinomycin when expressed in the chloroplast. Although the mutant is able to grow phototrophically, photosystem II function and cell growth are impaired under stress conditions such as high light intensity and diminished chloroplast protein synthesis induced by spectinomycin. Use of an antibody generated against the ycf8 product has revealed that this hydrophobic polypeptide is associated with photosystem II, based on its severely reduced levels in various photosystem II-deficient mutants and on its copurification with photosystem II. This protein, therefore, appears to be (i) a novel photosystem II subunit and (ii) required for maintaining optimal photosystem II activity under adverse growth conditions.

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Selected References

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  1. Barber J., Andersson B. Too much of a good thing: light can be bad for photosynthesis. Trends Biochem Sci. 1992 Feb;17(2):61–66. doi: 10.1016/0968-0004(92)90503-2. [DOI] [PubMed] [Google Scholar]
  2. Büschlen S., Choquet Y., Kuras R., Wollman F. A. Nucleotide sequences of the continuous and separated petA, petB and petD chloroplast genes in Chlamydomonas reinhardtii. FEBS Lett. 1991 Jun 24;284(2):257–262. doi: 10.1016/0014-5793(91)80698-3. [DOI] [PubMed] [Google Scholar]
  3. Choquet Y., Goldschmidt-Clermont M., Girard-Bascou J., Kück U., Bennoun P., Rochaix J. D. Mutant phenotypes support a trans-splicing mechanism for the expression of the tripartite psaA gene in the C. reinhardtii chloroplast. Cell. 1988 Mar 25;52(6):903–913. doi: 10.1016/0092-8674(88)90432-1. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Goldschmidt-Clermont M., Choquet Y., Girard-Bascou J., Michel F., Schirmer-Rahire M., Rochaix J. D. A small chloroplast RNA may be required for trans-splicing in Chlamydomonas reinhardtii. Cell. 1991 Apr 5;65(1):135–143. doi: 10.1016/0092-8674(91)90415-u. [DOI] [PubMed] [Google Scholar]
  6. Goldschmidt-Clermont M., Girard-Bascou J., Choquet Y., Rochaix J. D. Trans-splicing mutants of Chlamydomonas reinhardtii. Mol Gen Genet. 1990 Sep;223(3):417–425. doi: 10.1007/BF00264448. [DOI] [PubMed] [Google Scholar]
  7. Goldschmidt-Clermont M. Transgenic expression of aminoglycoside adenine transferase in the chloroplast: a selectable marker of site-directed transformation of chlamydomonas. Nucleic Acids Res. 1991 Aug 11;19(15):4083–4089. doi: 10.1093/nar/19.15.4083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hallick R. B., Hong L., Drager R. G., Favreau M. R., Monfort A., Orsat B., Spielmann A., Stutz E. Complete sequence of Euglena gracilis chloroplast DNA. Nucleic Acids Res. 1993 Jul 25;21(15):3537–3544. doi: 10.1093/nar/21.15.3537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hiratsuka J., Shimada H., Whittier R., Ishibashi T., Sakamoto M., Mori M., Kondo C., Honji Y., Sun C. R., Meng B. Y. The complete sequence of the rice (Oryza sativa) chloroplast genome: intermolecular recombination between distinct tRNA genes accounts for a major plastid DNA inversion during the evolution of the cereals. Mol Gen Genet. 1989 Jun;217(2-3):185–194. doi: 10.1007/BF02464880. [DOI] [PubMed] [Google Scholar]
  10. Ikeuchi M., Eggers B., Shen G. Z., Webber A., Yu J. J., Hirano A., Inoue Y., Vermaas W. Cloning of the psbK gene from Synechocystis sp. PCC 6803 and characterization of photosystem II in mutants lacking PSII-K. J Biol Chem. 1991 Jun 15;266(17):11111–11115. [PubMed] [Google Scholar]
  11. Jensen K. H., Herrin D. L., Plumley F. G., Schmidt G. W. Biogenesis of photosystem II complexes: transcriptional, translational, and posttranslational regulation. J Cell Biol. 1986 Oct;103(4):1315–1325. doi: 10.1083/jcb.103.4.1315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Johnson C. H., Schmidt G. W. The psbB gene cluster of the Chlamydomonas reinhardtii chloroplast: sequence and transcriptional analyses of psbN and psbH. Plant Mol Biol. 1993 Jul;22(4):645–658. doi: 10.1007/BF00047405. [DOI] [PubMed] [Google Scholar]
  13. Kuchka M. R., Mayfield S. P., Rochaix J. D. Nuclear mutations specifically affect the synthesis and/or degradation of the chloroplast-encoded D2 polypeptide of photosystem II in Chlamydomonas reinhardtii. EMBO J. 1988 Feb;7(2):319–324. doi: 10.1002/j.1460-2075.1988.tb02815.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mayfield S. P., Rahire M., Frank G., Zuber H., Rochaix J. D. Expression of the nuclear gene encoding oxygen-evolving enhancer protein 2 is required for high levels of photosynthetic oxygen evolution in Chlamydomonas reinhardtii. Proc Natl Acad Sci U S A. 1987 Feb;84(3):749–753. doi: 10.1073/pnas.84.3.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Monod C., Goldschmidt-Clermont M., Rochaix J. D. Accumulation of chloroplast psbB RNA requires a nuclear factor in Chlamydomonas reinhardtii. Mol Gen Genet. 1992 Feb;231(3):449–459. doi: 10.1007/BF00292715. [DOI] [PubMed] [Google Scholar]
  16. Rochaix J. D., Kuchka M., Mayfield S., Schirmer-Rahire M., Girard-Bascou J., Bennoun P. Nuclear and chloroplast mutations affect the synthesis or stability of the chloroplast psbC gene product in Chlamydomonas reinhardtii. EMBO J. 1989 Apr;8(4):1013–1021. doi: 10.1002/j.1460-2075.1989.tb03468.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  18. Shinozaki K., Ohme M., Tanaka M., Wakasugi T., Hayashida N., Matsubayashi T., Zaita N., Chunwongse J., Obokata J., Yamaguchi-Shinozaki K. The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression. EMBO J. 1986 Sep;5(9):2043–2049. doi: 10.1002/j.1460-2075.1986.tb04464.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  20. Takahashi Y., Goldschmidt-Clermont M., Soen S. Y., Franzén L. G., Rochaix J. D. Directed chloroplast transformation in Chlamydomonas reinhardtii: insertional inactivation of the psaC gene encoding the iron sulfur protein destabilizes photosystem I. EMBO J. 1991 Aug;10(8):2033–2040. doi: 10.1002/j.1460-2075.1991.tb07733.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Weeks D. P., Beerman N., Griffith O. M. A small-scale five-hour procedure for isolating multiple samples of CsCl-purified DNA: application to isolations from mammalian, insect, higher plant, algal, yeast, and bacterial sources. Anal Biochem. 1986 Feb 1;152(2):376–385. doi: 10.1016/0003-2697(86)90423-9. [DOI] [PubMed] [Google Scholar]
  22. de Vitry C., Olive J., Drapier D., Recouvreur M., Wollman F. A. Posttranslational events leading to the assembly of photosystem II protein complex: a study using photosynthesis mutants from Chlamydomonas reinhardtii. J Cell Biol. 1989 Sep;109(3):991–1006. doi: 10.1083/jcb.109.3.991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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