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Plant Molecular Biology

, Volume 39, Issue 1, pp 1–10 | Cite as

Cloning and characterization of the nuclear AC115 gene of Chlamydomonas reinhardtii

  • Pongsak Rattanachaikunsopon
  • Cristen Rosch
  • Michael R. Kuchka
Article

Abstract

The nuclear ac115 mutant of Chlamydomonas reinhardtii is specifically blocked in the synthesis of the chloroplast encoded D2 protein of the photosystem II reaction center at a point after translation initiation. Here, we report the identification of the AC115 gene through complementation rescue of the ac115 mutant strain, using an indexed cosmid library of Chlamydomonas genomic DNA. AC115 is a small, novel, intronless nuclear gene which encodes a protein of 113 amino acids. The amino terminal end of the Ac115 protein is rich in basic amino acids and has features which resemble a chloroplast transit sequence. A hydrophobic stretch of amino acids at the protein's carboxyl terminus is sufficiently large to be a membrane spanning or a protein/protein interaction domain. Various models are discussed to account for the mechanism by which Ac115p works in D2 synthesis. The ac115 mutant allele was sequenced and determined to be an A-to-T transversion at the first position of the fourth codon of the coding sequence. This mutation changes an AAG codon to a TAG nonsense codon and results in a null phenotype.

Chlamydomonas chloroplast D2 protein nuclear gene photosystem II 

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References

  1. 1.
    Andreasson L-E, Vangaard T: Electron transport in photosystem I and II. Plant Mol Biol 39: 379–411 (1988).Google Scholar
  2. 2.
    Anbudurai PR, Pakrasi HB: Mutational analysis of the psbL protein of photosystem II in the cyanobacterium Synechocystis sp. PCC6803. Z Naturforsch 48c: 267–274 (1993).Google Scholar
  3. 3.
    Boynton JE, Gillham NW, Harris EH, Hosler JP, Johnson AM, Jones AR, Randolph-Anderson BL, Robertson D, Klein TM, Shark KB, Sanford JC: Chloroplast transformation in Chlamydomonas with high velocity microprojectiles. Science 240: 1534–1538 (1988).Google Scholar
  4. 4.
    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 52: 901–912 (1988).Google Scholar
  5. 5.
    Cline K, Henry R: Import and routing of nucleus-encoded chloroplast proteins. Annu Rev Cell Devel Biol 12: 1–26 (1996).Google Scholar
  6. 6.
    Covello PS, Gray MW: Silent mitochondrial and active nuclear genes for subunit 2 of cytochrome c oxidase (cox2) in soybean: evidence for RNA-mediated gene transfer. EMBO J 11: 3815–3820 (1992).Google Scholar
  7. 7.
    DeVitry C, Diner BA, Popot J-L: Photosystem II particles from Chlamydomonas reinhardtii. J Biol Chem 266: 16614–16621 (1991).Google Scholar
  8. 8.
    Erickson JM, Delepelaire P, Rochaix, J-D: Analysis of genes encoding two photosystem II proteins of the 30–34 kD size class. In: Steinback KE (ed) Molecular Biology of the Photosynthetic Apparatus, pp. 53–66. Cold Spring Harbor Press, Cold Spring Harbor, NY (1985).Google Scholar
  9. 9.
    Erickson JM, Rahire M, Malnoe P, Girard-Bascou J, Pierre Y, Bennoun P, Rochaix J-D: Lack of D2 protein in a Chlamydomonas reinhardtii psbD mutant affects photosystem II stability and D1 expression. EMBO J 5: 1745-1754 (1986).Google Scholar
  10. 10.
    Goldschmidt-Clermont M, Rahire M: Sequence, evolution and differential expression of the two genes encoding variant small subunits of ribulose bisphosphate carboxylase/oxygenase in Chlamydomonas reinhardtii. J Mol Biol 191: 421–432 (1986).Google Scholar
  11. 11.
    Goldschmidt-Clermont M, Girard-Bascou J, Choquet Y, Rochaix J-D: Trans-splicing mutants of Chlamydomonas reinhardtii. Mol Gen Genet 223: 417–425 (1990).Google Scholar
  12. 12.
    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 Chamydomonas reinhardtii. Cell 65: 135–143 (1991).Google Scholar
  13. 13.
    Goldschmidt-Clermont M: Coordination of nuclear and chloroplast gene expression in plant cells. Int Rev Cytol 177: 115–180 (1998).Google Scholar
  14. 14.
    Goodenough UW, Levine RP: Chloroplast ultrastructure in mutant strains of Chlamydomonas reinhardtii lacking components of the photosynthetic apparatus. Plant Physiol 44: 990–1000 (1969).Google Scholar
  15. 15.
    Gorman DS, Levine RP: Cytochrome f and plastocyanin: their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardi. Proc Natl Acad Sci USA 54: 1665–1669 (1965).Google Scholar
  16. 16.
    Harris E: The Chlamydomonas Sourcebook: A Comprehensive Guide to Biology and Laboratory Use. Academic Press, San Diego, CA (1989).Google Scholar
  17. 17.
    Heidecker G, Messing J: Structural analysis of plant genes. Annu Rev Plant Physiol 37: 439–466 (1986).Google Scholar
  18. 18.
    Herrin D, Michaels A: The chloroplast 32 kDa protein is synthesized on thylakoid-bound ribosomes in Chlamydomonas reinhardtii. FEBS Lett 184: 90–95 (1985).Google Scholar
  19. 19.
    Hyams J, Davies CR: The induction and characterization of cell wall mutants of Chlamydomonas reinhardi. Mutat Res 14: 381–389 (1972).Google Scholar
  20. 20.
    Infante A, Lo S, Hall JL: A Chlamydomonas genomic library in yeast artificial chromosomes. Genetics 141: 87–93 (1995).Google Scholar
  21. 21.
    Irrgang KD, Shi LX, Funk C, Schroder WP: A nuclearencoded subunit of the photosystem II reaction center. J Biol Chem 270: 17588–17593 (1995).Google Scholar
  22. 22.
    Kapazoglou A, Sagliocco F, Dure L: PSII-T, a new nuclear encoded lumenal protein from photosystem II. Targeting and processing in isolated chloroplasts. J Biol Chem 270: 12197–12202 (1995).Google Scholar
  23. 23.
    Kim J, Gamble Klein P, Mullet JE: Vir-115 gene product is required to stabilize D1 translation intermediates in chloroplasts. Plant Mol Biol 25: 459–467 (1994).Google Scholar
  24. 24.
    Kindle KL: High-frequency nuclear transformation of Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 87: 1228–1232 (1990).Google Scholar
  25. 25.
    Kindle KL, Sodeinde OA: Nuclear and chloroplast transformation in Chlamydomonas reinhardtii: strategies for genetic manipulation and gene expression. J Appl Phycol 6: 231–238 (1994).Google Scholar
  26. 26.
    Kuchka MR, Mayfield SP, 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 7: 319–324 (1988).Google Scholar
  27. 27.
    Kuchka MR, Goldschmidt-Clermont M, vanDillewijn J, Rochaix J-D: Mutation at the Chlamydomonas nuclear NAC2 locus specifically affects stability of the chloroplast psbD transcript encoding polypeptide D2 of PSII. Cell 58: 869–876 (1989).Google Scholar
  28. 28.
    Levine RP, Volkmann D: Mutants with impaired photosynthesis in Chlamydomonas reinhardtii. Biochem Biophys Res Commun 6: 264–269 (1961).Google Scholar
  29. 29.
    Levine RP, Goodenough UW: The genetics of photosynthesis and the chloroplast in Chlamydomonas reinhardi. Annu Rev Genet 4: 397–408 (1970).Google Scholar
  30. 30.
    Lorkovic ZJ, Schroder WP, Pakrasi IHB, Irrgang KD, Herrman RG, Oelmuller R: Molecular characterization of PsbW, a nuclear-encoded component of the photosystem II reaction center complex in spinach. Proc Natl Acad Sci USA 92: 8930–8934 (1995).Google Scholar
  31. 31.
    Maniatis T, Fritsch EF, and Sambrook J: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).Google Scholar
  32. 32.
    Mattoo AK, Marder JB, Edelman E: Dynamics of the photosystem II reaction center. Cell 56: 241–246 (1989).Google Scholar
  33. 33.
    Merchant S, Bogorad L: The Cu(II)-repressible plastidic cytochrome c: cloning and sequence of a complementary DNA for the pre-apoprotein. J Biol Chem 262: 9062–9067 (1987).Google Scholar
  34. 34.
    Nanba O, Satoh K: Isolation of a photosystem II reaction center consisting of D-1 and D-2 polypeptides and cytochrome b559. Proc Natl Acad Sci USA 84: 109–112 (1987).Google Scholar
  35. 35.
    Nickelson J, vanDillewijn, J, Rahire M, Rochaix J-D: Determinants for stability of the chloroplast psbD RNA are located within its short leader region in Chlamydomonas reinhardtii. EMBO J 13: 3182–3191 (1994).Google Scholar
  36. 36.
    Nugent JM, Palmer JD: RNA-mediated transfer of the gene coxII from mitochondrion to the nucleus during flowering plant evolution. Cell 66: 473–481 (1991).Google Scholar
  37. 37.
    Palmer J: Plastid chromosomes: structure and evolution. In: Bogorad L, Vasil IK (eds) The Molecular Biology of Plastids, pp. 5–53. Academic Press, New York (1991).Google Scholar
  38. 38.
    Purton S, Rochaix J-D: Complementation of a Chlamydomonas reinhardtii mutant using a genomic cosmid library. Plant Mol Biol 24: 533–537 (1994).Google Scholar
  39. 39.
    Rochaix J-D, Mayfield S, Goldschmidt-Clermont M, Erickson J: Molecular biology of Chlamydomonas. In: Shaw CH (ed) Plant Molecular Biology: A Practical Approach, pp. 253–277. IRL Press, New York (1988).Google Scholar
  40. 40.
    Silflow CD, Chisholm RL, Conner TW, Ranum LPW: The two alpha-tubulin genes of Chlamydomonas reinhardtii code for slightly different proteins. Mol Cell Biol 5: 2389–2398 (1985).Google Scholar
  41. 41.
    Smillie RM, Levine RP: The photosynthetic electron transport chain of Chlamydomonas reinhardtii. J Biol Chem 238: 4058–4062 (1962).Google Scholar
  42. 42.
    Sodeinde OA, Kindle KL: Homologous recombination in the nuclear genome of Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 90: 9199–9203 (1993).Google Scholar
  43. 43.
    Thorsness PE, Weber ER: Escape and migration of nucleic acids between chloroplasts, mitochondria, and the nucleus. Int Rev Cytol 165: 207–234 (1996).Google Scholar
  44. 44.
    Wu HY, Kuchka MR: A nuclear suppressor overcomes defects in the synthesis of the chloroplast psbD gene product caused bymutations in two distinct nuclear genes of Chlamydomonas. Curr Genet 27: 263–269 (1995).Google Scholar
  45. 45.
    Youngblom J, Schloss JA, Silflow CD: The two beta-tubulin gene of Chlamydomonas reinhardtii code for identical proteins. Mol Cell Biol 4: 2686–2696 (1984).Google Scholar
  46. 46.
    Zhang H, Herman PL, Weeks DP: Gene isolation through genomic complementation using an indexed library of Chlamydomonas reinhardtii DNA. Plant Mol Biol 24: 633–672 (1994).Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Pongsak Rattanachaikunsopon
    • 2
  • Cristen Rosch
    • 1
  • Michael R. Kuchka
    • 1
  1. 1.Department of Biological SciencesLehigh UniversityBethlehemUSA
  2. 2.Ubon Ratchathani University, UbonRatchathaniThailand

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