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. 1986 Nov 1;103(5):1837–1845. doi: 10.1083/jcb.103.5.1837

Regulation of genes encoding the large subunit of ribulose-1,5- bisphosphate carboxylase and the photosystem II polypeptides D-1 and D- 2 during the cell cycle of Chlamydomonas reinhardtii

PMCID: PMC2114377  PMID: 3536953

Abstract

Synthesis of the major chloroplast proteins is temporally regulated in light-dark-synchronized Chlamydomonas cells. We have used cloned chloroplast DNA probes, and in vitro and in vivo protein synthesis to examine the cell cycle regulation of photosystem II polypeptides D-1 and D-2, and the large subunit of ribulose-1,5-bisphosphate carboxylase (RuBPCase LS). Synthesis and accumulation of D-1 and D-2 mRNAs occurs during the first half of the light period (G1), correlating with increasing synthesis of the polypeptides. Rifampicin, added immediately before the light period, inhibited the normal increase in D-1, D-2 polypeptide synthesis. During the dark period D-1, D-2 mRNAs persist at high levels despite reduced rates of mRNA synthesis and translation during this period. Cell-free translation analyses indicate that the D- 1 mRNA present during the dark period is efficient at directing synthesis of the D-1 precursor in vitro. We conclude that expression of the psbA (D-1) and psbD (D-2) genes are regulated primarily at the transcriptional level during the light-induction period but at the translational level for the remainder of the cell cycle. Transcripts of the RuBPCase LS gene (rbcL) are also found at high levels during the light and dark periods but, unlike D-1 and D-2, LS mRNA levels do not increase until the last half of the light period and measurable synthesis and accumulation of this mRNA occurs during the dark. Furthermore, induction of LS polypeptide synthesis during the light period is insensitive to rifampicin. We conclude that LS production is regulated primarily at the translational level during the cell cycle.

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Selected References

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  1. Ares M., Jr, Howell S. H. Cell cycle stage-specific accumulation of mRNAs encoding tubulin and other polypeptides in Chlamydomonas. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5577–5581. doi: 10.1073/pnas.79.18.5577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Armstrong J. J., Moll B., Surzycki S. J., Levine R. P. Genetic transcription and translation specifying chloroplast components in Chlamydomonas reinhardi. Biochemistry. 1971 Feb 16;10(4):692–701. doi: 10.1021/bi00780a022. [DOI] [PubMed] [Google Scholar]
  3. Baker E. J., Schloss J. A., Rosenbaum J. L. Rapid changes in tubulin RNA synthesis and stability induced by deflagellation in Chlamydomonas. J Cell Biol. 1984 Dec;99(6):2074–2081. doi: 10.1083/jcb.99.6.2074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beck D. P., Levine R. P. Synthesis of chloroplast membrane polypeptides during synchronous growth of Chlamydomonas reinhardtii. J Cell Biol. 1974 Dec;63(3):759–772. doi: 10.1083/jcb.63.3.759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Berry J. O., Nikolau B. J., Carr J. P., Klessig D. F. Transcriptional and post-transcriptional regulation of ribulose 1,5-bisphosphate carboxylase gene expression in light- and dark-grown amaranth cotyledons. Mol Cell Biol. 1985 Sep;5(9):2238–2246. doi: 10.1128/mcb.5.9.2238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chamberlain J. P. Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate. Anal Biochem. 1979 Sep 15;98(1):132–135. doi: 10.1016/0003-2697(79)90716-4. [DOI] [PubMed] [Google Scholar]
  8. Chiang K. S., Sueoka N. Replication of chloroplast DNA in Chlamydomonas reinhardi during vegetative cell cycle: its mode and regulation. Proc Natl Acad Sci U S A. 1967 May;57(5):1506–1513. doi: 10.1073/pnas.57.5.1506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chua N. H., Blomberg F. Immunochemical studies of thylakoid membrane polypeptides from spinach and Chlamydomonas reinhardtii. A modified procedure for crossed immunoelectrophoresis of dodecyl sulfate.protein complexes. J Biol Chem. 1979 Jan 10;254(1):215–223. [PubMed] [Google Scholar]
  10. Chua N. H., Gillham N. W. The sites of synthesis of the principal thylakoid membrane polypeptides in Chlamydomonas reinhardtii. J Cell Biol. 1977 Aug;74(2):441–452. doi: 10.1083/jcb.74.2.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dallman T., Ares M., Jr, Howell S. H. Analysis of transcription during the cell cycle in toluenized Chlamydomonas reinhardi cells. Mol Cell Biol. 1983 Aug;3(8):1537–1539. doi: 10.1128/mcb.3.8.1537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Delepelaire P., Chua N. H. Electrophoretic purification of chlorophyll a/b-protein complexes from Chlamydomonas reinhardtii and spinach and analysis of their polypeptide compositions. J Biol Chem. 1981 Sep 10;256(17):9300–9307. [PubMed] [Google Scholar]
  13. Delepelaire P. Partial characterization of the biosynthesis and integration of the Photosystem II reaction centers in the thylakoid membrane of Chlamydomonas reinhardtii. EMBO J. 1984 Apr;3(4):701–706. doi: 10.1002/j.1460-2075.1984.tb01872.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dron M., Rahire M., Rochaix J. D. Sequence of the chloroplast DNA region of Chlamydomonas reinhardii containing the gene of the large subunit of ribulose bisphosphate carboxylase and parts of its flanking genes. J Mol Biol. 1982 Dec 25;162(4):775–793. doi: 10.1016/0022-2836(82)90547-2. [DOI] [PubMed] [Google Scholar]
  15. Erickson J. M., Rahire M., Rochaix J. D. Chlamydomonas reinhardii gene for the 32 000 mol. wt. protein of photosystem II contains four large introns and is located entirely within the chloroplast inverted repeat. EMBO J. 1984 Dec 1;3(12):2753–2762. doi: 10.1002/j.1460-2075.1984.tb02206.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fromm H., Devic M., Fluhr R., Edelman M. Control of psbA gene expression: in mature Spirodela chloroplasts light regulation of 32-kd protein synthesis is independent of transcript level. EMBO J. 1985 Feb;4(2):291–295. doi: 10.1002/j.1460-2075.1985.tb03628.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Herrin D., Michaels A. In vitro synthesis and assembly of the peripheral subunits of coupling factor CF1 (alpha and beta) by thylakoid-bound ribosomes. Arch Biochem Biophys. 1985 Feb 15;237(1):224–236. doi: 10.1016/0003-9861(85)90273-5. [DOI] [PubMed] [Google Scholar]
  18. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  19. Howell S. H., Blaschko W. J., Drew C. M. Inhibitor effects during the cell cycle in Chlamydomonas reinhardtii. Determination of transition points in asynchronous cultures. J Cell Biol. 1975 Oct;67(1):126–135. doi: 10.1083/jcb.67.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Howell S. H., Naliboff J. A. Conditional mutants in Chlamydomonas reinhardtii blocked in the vegetative cell cycle. I. An analysis of cell cycle block points. J Cell Biol. 1973 Jun;57(3):760–772. doi: 10.1083/jcb.57.3.760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Howell S. H., Posakony J. W., Hill K. R. The cell cycle program of polypeptide labeling in Chlamydomonas reinhardtii. J Cell Biol. 1977 Feb;72(2):223–241. doi: 10.1083/jcb.72.2.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Howell S. H., Walker L. L. Transcription of the nuclear and chloroplast genomes during the vegetative cell cycle in Chlamydomonas reinhardi. Dev Biol. 1977 Mar;56(1):11–23. doi: 10.1016/0012-1606(77)90151-8. [DOI] [PubMed] [Google Scholar]
  23. Inamine G., Nash B., Weissbach H., Brot N. Light regulation of the synthesis of the large subunit of ribulose-1,5-bisphosphate carboxylase in peas: Evidence for translational control. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5690–5694. doi: 10.1073/pnas.82.17.5690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Janero D. R., Barrnett R. Thylakoid membrane biogenesis in Chlamydomonas reinhardtii 137+. II. Cell-cycle variations in the synthesis and assembly of pigment. J Cell Biol. 1982 May;93(2):411–416. doi: 10.1083/jcb.93.2.411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Janero D. R., Barrnett R. Thylakoid membrane biogenesis in Chlamydomonas reinhardtii 137+: cell cycle variations in the synthesis and assembly of polar glycerolipid. J Cell Biol. 1981 Oct;91(1):126–134. doi: 10.1083/jcb.91.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kafatos F. C., Jones C. W., Efstratiadis A. Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. Nucleic Acids Res. 1979 Nov 24;7(6):1541–1552. doi: 10.1093/nar/7.6.1541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  29. 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]
  30. Miller M. E., Jurgenson J. E., Reardon E. M., Price C. A. Plastid translation in organello and in vitro during light-induced development in Euglena. J Biol Chem. 1983 Dec 10;258(23):14478–14484. [PubMed] [Google Scholar]
  31. Nelson T., Harpster M. H., Mayfield S. P., Taylor W. C. Light-regulated gene expression during maize leaf development. J Cell Biol. 1984 Feb;98(2):558–564. doi: 10.1083/jcb.98.2.558. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Palmer J. D., Edwards H., Jorgensen R. A., Thompson W. F. Novel evolutionary variation in transcription and location of two chloroplast genes. Nucleic Acids Res. 1982 Nov 11;10(21):6819–6832. doi: 10.1093/nar/10.21.6819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rosner A., Jakob K. M., Gressel J., Sagher D. The early synthesis and possible function of a 0.5 X 10(6) Mr RNA after transfer of dark-grown Spirodela plants to light. Biochem Biophys Res Commun. 1975 Nov 3;67(1):383–391. doi: 10.1016/0006-291x(75)90327-7. [DOI] [PubMed] [Google Scholar]
  34. Shepherd H. S., Ledoigt G., Howell S. H. Regulation of light-harvesting chlorophyll-binding protein (LHCP) mRNA accumulation during the cell cycle in Chlamydomonas reinhardi. Cell. 1983 Jan;32(1):99–107. doi: 10.1016/0092-8674(83)90500-7. [DOI] [PubMed] [Google Scholar]
  35. Shinozaki K., Sasaki Y., Sakihama T., Kamikubo T. Coordinate light-induction of two mRNAs, encoded in nuclei and chloroplasts, of ribulose 1,5-bisphosphate carboxylase/oxygenase. FEBS Lett. 1982 Jul 19;144(1):73–76. doi: 10.1016/0014-5793(82)80571-1. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Spudich J. L., Sager R. Regulation of the Chlamydomonas cell cycle by light and dark. J Cell Biol. 1980 Apr;85(1):136–145. doi: 10.1083/jcb.85.1.136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Stitt M., Lilley R. M., Heldt H. W. Adenine nucleotide levels in the cytosol, chloroplasts, and mitochondria of wheat leaf protoplasts. Plant Physiol. 1982 Oct;70(4):971–977. doi: 10.1104/pp.70.4.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Surzycki S. J., Rochaix J. D. Transcriptional mapping of ribosomal RNA genes of the chloroplast and nucleus of Chlamydomonas reinhardi. J Mol Biol. 1971 Nov 28;62(1):89–109. doi: 10.1016/0022-2836(71)90133-1. [DOI] [PubMed] [Google Scholar]

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