Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1997 Oct 15;16(20):6095–6104. doi: 10.1093/emboj/16.20.6095

The chloroplast ycf3 and ycf4 open reading frames of Chlamydomonas reinhardtii are required for the accumulation of the photosystem I complex.

E Boudreau 1, Y Takahashi 1, C Lemieux 1, M Turmel 1, J D Rochaix 1
PMCID: PMC1326293  PMID: 9321389

Abstract

The chloroplast genes ycf3 and ycf4 from the green alga Chlamydomonas reinhardtii have been characterized. The deduced amino acid sequences of Ycf4 (197 residues) and Ycf3 (172 residues) display 41-52% and 64-78% sequence identity, respectively, with their homologues from algae, land plants and cyanobacteria. In C. reinhardtii, ycf4 and ycf3 are co-transcribed as members of the rps9-ycf4-ycf3-rps18 polycistronic transcriptional unit into RNAs of 8.0 kb and 3.0 kb corresponding to the entire unit and to rps9-ycf4-ycf3, respectively. Using biolistic transformation, ycf4 and ycf3 were disrupted with a chloroplast selectable marker cassette. Transformants lacking ycf4 or ycf3 were unable to grow photoautotrophically and were deficient in photosystem I activity. Western blot analysis showed that the photosystem I (PSI) complex does not accumulate stably in thylakoid membranes of these transformants. Ycf4 and Ycf3 were localized on thylakoid membranes but not stably associated with the PSI complex and accumulated to wild-type levels in mutants lacking PSI. RNA blot hybridizations showed that transcripts of psaA, psaB and psaC accumulate normally in these mutants and use of chimeric reporter genes revealed that Ycf3 is not required for initiation of translation of psaA and psaB mRNA. Our results indicate that Ycf3 and Ycf4 are required for stable accumulation of the PSI complex.

Full Text

The Full Text of this article is available as a PDF (528.4 KB).

Selected References

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

  1. Altamura N., Capitanio N., Bonnefoy N., Papa S., Dujardin G. The Saccharomyces cerevisiae OXA1 gene is required for the correct assembly of cytochrome c oxidase and oligomycin-sensitive ATP synthase. FEBS Lett. 1996 Mar 11;382(1-2):111–115. doi: 10.1016/0014-5793(96)00165-2. [DOI] [PubMed] [Google Scholar]
  2. Baldauf S. L., Manhart J. R., Palmer J. D. Different fates of the chloroplast tufA gene following its transfer to the nucleus in green algae. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5317–5321. doi: 10.1073/pnas.87.14.5317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boudreau E., Otis C., Turmel M. Conserved gene clusters in the highly rearranged chloroplast genomes of Chlamydomonas moewusii and Chlamydomonas reinhardtii. Plant Mol Biol. 1994 Feb;24(4):585–602. doi: 10.1007/BF00023556. [DOI] [PubMed] [Google Scholar]
  4. Boudreau E., Turmel M. Extensive gene rearrangements in the chloroplast DNAs of Chlamydomonas species featuring multiple dispersed repeats. Mol Biol Evol. 1996 Jan;13(1):233–243. doi: 10.1093/oxfordjournals.molbev.a025560. [DOI] [PubMed] [Google Scholar]
  5. Boudreau E., Turmel M. Gene rearrangements in Chlamydomonas chloroplast DNAs are accounted for by inversions and by the expansion/contraction of the inverted repeat. Plant Mol Biol. 1995 Jan;27(2):351–364. doi: 10.1007/BF00020189. [DOI] [PubMed] [Google Scholar]
  6. Boudreau E., Turmel M., Goldschmidt-Clermont M., Rochaix J. D., Sivan S., Michaels A., Leu S. A large open reading frame (orf1995) in the chloroplast DNA of Chlamydomonas reinhardtii encodes an essential protein. Mol Gen Genet. 1997 Feb 20;253(5):649–653. doi: 10.1007/s004380050368. [DOI] [PubMed] [Google Scholar]
  7. Buchheim M. A., Lemieux C., Otis C., Gutell R. R., Chapman R. L., Turmel M. Phylogeny of the Chlamydomonadales (Chlorophyceae): a comparison of ribosomal RNA gene sequences from the nucleus and the chloroplast. Mol Phylogenet Evol. 1996 Apr;5(2):391–402. doi: 10.1006/mpev.1996.0034. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Fischer N., Stampacchia O., Redding K., Rochaix J. D. Selectable marker recycling in the chloroplast. Mol Gen Genet. 1996 Jun 12;251(3):373–380. doi: 10.1007/BF02172529. [DOI] [PubMed] [Google Scholar]
  10. Gantt J. S., Baldauf S. L., Calie P. J., Weeden N. F., Palmer J. D. Transfer of rpl22 to the nucleus greatly preceded its loss from the chloroplast and involved the gain of an intron. EMBO J. 1991 Oct;10(10):3073–3078. doi: 10.1002/j.1460-2075.1991.tb07859.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Girard-Bascou J., Choquet Y., Schneider M., Delosme M., Dron M. Characterization of a chloroplast mutation in the psaA2 gene of Chlamydomonas reinhardtii. Curr Genet. 1987;12(7):489–495. doi: 10.1007/BF00419557. [DOI] [PubMed] [Google Scholar]
  12. Glerum D. M., Koerner T. J., Tzagoloff A. Cloning and characterization of COX14, whose product is required for assembly of yeast cytochrome oxidase. J Biol Chem. 1995 Jun 30;270(26):15585–15590. doi: 10.1074/jbc.270.26.15585. [DOI] [PubMed] [Google Scholar]
  13. Goebl M., Yanagida M. The TPR snap helix: a novel protein repeat motif from mitosis to transcription. Trends Biochem Sci. 1991 May;16(5):173–177. doi: 10.1016/0968-0004(91)90070-c. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Hippler M., Drepper F., Farah J., Rochaix J. D. Fast electron transfer from cytochrome c6 and plastocyanin to photosystem I of Chlamydomonas reinhardtii requires PsaF. Biochemistry. 1997 May 27;36(21):6343–6349. doi: 10.1021/bi970082c. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Jacobsen S. E., Binkowski K. A., Olszewski N. E. SPINDLY, a tetratricopeptide repeat protein involved in gibberellin signal transduction in Arabidopsis. Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):9292–9296. doi: 10.1073/pnas.93.17.9292. [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. Lamb J. R., Tugendreich S., Hieter P. Tetratrico peptide repeat interactions: to TPR or not to TPR? Trends Biochem Sci. 1995 Jul;20(7):257–259. doi: 10.1016/s0968-0004(00)89037-4. [DOI] [PubMed] [Google Scholar]
  20. Leonhard K., Herrmann J. M., Stuart R. A., Mannhaupt G., Neupert W., Langer T. AAA proteases with catalytic sites on opposite membrane surfaces comprise a proteolytic system for the ATP-dependent degradation of inner membrane proteins in mitochondria. EMBO J. 1996 Aug 15;15(16):4218–4229. [PMC free article] [PubMed] [Google Scholar]
  21. Lubben T. H., Donaldson G. K., Viitanen P. V., Gatenby A. A. Several proteins imported into chloroplasts form stable complexes with the GroEL-related chloroplast molecular chaperone. Plant Cell. 1989 Dec;1(12):1223–1230. doi: 10.1105/tpc.1.12.1223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Madueno F., Napier J. A., Gray J. C. Newly Imported Rieske Iron-Sulfur Protein Associates with Both Cpn60 and Hsp70 in the Chloroplast Stroma. Plant Cell. 1993 Dec;5(12):1865–1876. doi: 10.1105/tpc.5.12.1865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Malnoë P., Mayfield S. P., Rochaix J. D. Comparative analysis of the biogenesis of photosystem II in the wild-type and Y-1 mutant of Chlamydomonas reinhardtii. J Cell Biol. 1988 Mar;106(3):609–616. doi: 10.1083/jcb.106.3.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mannan R. M., Whitmarsh J., Nyman P., Pakrasi H. B. Directed mutagenesis of an iron-sulfur protein of the photosystem I complex in the filamentous cyanobacterium Anabaena variabilis ATCC 29413. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10168–10172. doi: 10.1073/pnas.88.22.10168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mayfield S. P., Bennoun P., Rochaix J. D. Expression of the nuclear encoded OEE1 protein is required for oxygen evolution and stability of photosystem II particles in Chlamydomonas reinhardtii. EMBO J. 1987 Feb;6(2):313–318. doi: 10.1002/j.1460-2075.1987.tb04756.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mor T. S., Ohad I., Hirschberg J., Pakrasi H. B. An unusual organization of the genes encoding cytochrome b559 in Chlamydomonas reinhardtii: psbE and psbF genes are separately transcribed from different regions of the plastid chromosome. Mol Gen Genet. 1995 Mar 10;246(5):600–604. doi: 10.1007/BF00298966. [DOI] [PubMed] [Google Scholar]
  27. Ohad I., Siekevitz P., Palade G. E. Biogenesis of chloroplast membranes. II. Plastid differentiation during greening of a dark-grown algal mutant (Chlamydomonas reinhardi). J Cell Biol. 1967 Dec;35(3):553–584. doi: 10.1083/jcb.35.3.553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pakrasi H. B. Genetic analysis of the form and function of photosystem I and photosystem II. Annu Rev Genet. 1995;29:755–776. doi: 10.1146/annurev.ge.29.120195.003543. [DOI] [PubMed] [Google Scholar]
  29. Rochaix J. D. Post-transcriptional regulation of chloroplast gene expression in Chlamydomonas reinhardtii. Plant Mol Biol. 1996 Oct;32(1-2):327–341. doi: 10.1007/BF00039389. [DOI] [PubMed] [Google Scholar]
  30. Stampacchia O., Girard-Bascou J., Zanasco J. L., Zerges W., Bennoun P., Rochaix J. D. A nuclear-encoded function essential for translation of the chloroplast psaB mRNA in chlamydomonas. Plant Cell. 1997 May;9(5):773–782. doi: 10.1105/tpc.9.5.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Szczepaniak A., Huang D., Keenan T. W., Cramer W. A. Electrostatic destabilization of the cytochrome b6f complex in the thylakoid membrane. EMBO J. 1991 Oct;10(10):2757–2764. doi: 10.1002/j.1460-2075.1991.tb07824.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Takahashi Y., Matsumoto H., Goldschmidt-Clermont M., Rochaix J. D. Directed disruption of the Chlamydomonas chloroplast psbK gene destabilizes the photosystem II reaction center complex. Plant Mol Biol. 1994 Mar;24(5):779–788. doi: 10.1007/BF00029859. [DOI] [PubMed] [Google Scholar]
  34. Tzagoloff A., Yue J., Jang J., Paul M. F. A new member of a family of ATPases is essential for assembly of mitochondrial respiratory chain and ATP synthetase complexes in Saccharomyces cerevisiae. J Biol Chem. 1994 Oct 21;269(42):26144–26151. [PubMed] [Google Scholar]
  35. Wilde A., Härtel H., Hübschmann T., Hoffmann P., Shestakov S. V., Börner T. Inactivation of a Synechocystis sp strain PCC 6803 gene with homology to conserved chloroplast open reading frame 184 increases the photosystem II-to-photosystem I ratio. Plant Cell. 1995 May;7(5):649–658. doi: 10.1105/tpc.7.5.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Xie Z., Merchant S. The plastid-encoded ccsA gene is required for heme attachment to chloroplast c-type cytochromes. J Biol Chem. 1996 Mar 1;271(9):4632–4639. doi: 10.1074/jbc.271.9.4632. [DOI] [PubMed] [Google Scholar]
  37. Xu Q., Hoppe D., Chitnis V. P., Odom W. R., Guikema J. A., Chitnis P. R. Mutational analysis of photosystem I polypeptides in the cyanobacterium Synechocystis sp. PCC 6803. Targeted inactivation of psaI reveals the function of psaI in the structural organization of psaL. J Biol Chem. 1995 Jul 7;270(27):16243–16250. doi: 10.1074/jbc.270.27.16243. [DOI] [PubMed] [Google Scholar]
  38. Yu J., Smart L. B., Jung Y. S., Golbeck J., McIntosh L. Absence of PsaC subunit allows assembly of photosystem I core but prevents the binding of PsaD and PsaE in Synechocystis sp. PCC6803. Plant Mol Biol. 1995 Oct;29(2):331–342. doi: 10.1007/BF00043656. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES