Abstract
In Chlamydomonas reinhardtii, the chlorophyll a/b-binding proteins of photosystem II are encoded in the nucleus by a small family of genes. We have studied the expression of one gene, which we call cabII-1, in a green-in-the-dark strain, which can synthesize chlorophyll in the dark or light, and in a yellow-in-the-dark mutant strain, which is able to make chlorophyll only in the light. In light/dark synchronized cultures of both strains, cabII-1 mRNA abundance increases during the first 6 h of a 12-h light phase, remains high for several hours, then declines. A variety of illumination conditions have been used to analyze the cabII-1 mRNA increase: continuous or intermittent red, blue, or white light, with or without 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), an inhibitor of photosystem II. Our results suggest that light induces increased cabII-1 transcript abundance in two ways: 1) by virtue of its role in the light reactions of photosynthesis and 2) by a blue lightstimulated mechanism which is independent of photosynthesis.
We have also examined the role of acetate in regulating cabII-1 mRNA levels in the dark. In both green- and yellow-in-the-dark strains, 15 mM Na-acetate, added to synchronized cells in the dark, induces an increase in cabII-1 mRNA abundance with a temporal accumulation pattern very similar to that induced by continuous white light. We suggest that by providing an energy source, acetate stimulates cellular growth, cell cycle progression, and increased cabII-1 mRNA abundance. Interestingly, in cells exposed to light, acetate inhibits the light-induced increase in cabII-1 mRNA abundance by a mechanism which is not yet understood.
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References
Batschauer A, Mösinger E, Kreuz K, Dörr I, Apel K: The implications of a plastid-derived factor in the transcriptional control of nuclear genes encoding the light-harvesting chlorophyll a/b protein. Eur J Biochem 154: 625–634 (1986).
Bednarik DP, Hoober JK: Synthesis of chlorophyllide b from protochlorophyllide in Chlamydomonas reinhardtii y-1. Science 230: 450–453 (1985).
Chua N-H, Gillham NW: The sites of synthesis of the principal thylakoid membrane polypeptides in Chlamydomonas reinhardtii. J Cell Biol 74: 441–452 (1977).
Cordonnier MM, Greppin H, Pratt LH: Identification of a highly conserved domain on phytochrome from angiosperms to algae. Plant Physiol 80: 982–987 (1986).
Davies DR, Plaskitt A: Genetical and structural analyses of cell-wall formation in Chlamydomonas reinhardtii. Genet Res (Camb) 17: 33–43 (1971).
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 256: 9300–9307 (1981).
Eytan G, Ohad I: Biogenesis of chloroplast membranes. J Biol Chem 247: 122–129 (1972).
Fluhr R, Chua NH: Developmental regulation of two genes encoding ribulose-bisphosphate carboxylase small subunit in pea and transgenic petunia plants: Phytochrome response and blue-light induction. Proc Natl Acad Sci USA 83: 2358–2362 (1986).
Ford C, Wang WY: Instability of the y-1 locus of Chlamydomonas reinhardtii. Mol Gen Genet 187: 286–290 (1982).
Foster KW, Saranak J, Patel N, Zarilli G, Okabe M, Kline T, Nakanishi K: A rhodopsin is the functional photoreceptor for phototaxis in the unicellular eukaryote Chlamydomonas. Nature 311: 756–759 (1984).
Gallagher TF, Jenkins GI, Ellis RJ: Rapid modulation of transcription of nuclear genes encoding chloroplast proteins by light. FEBS Lett 186: 211–245 (1985).
Goldschmidt-Clermont M: The two genes for the small subunit of RuBP carboxylase/oxygenase are closely linked in Chlamydomonas reinhardtii. Plant Molec Biol 6: 13–21 (1986).
Hase E: Effect of blue light on greening in microalgae. In: Senger H (ed.) The Blue Light Syndrome. Springer-Verlag (1980) pp 512–525.
Hoober JK: Sites of synthesis of chloroplast membrane polypeptides in Chlamydomonas reinhardtii y-1. J Biol Chem 245: 4327–4334 (1970).
Hoober JH, Marks DB, Keller BJ, Margulies M: Regulation of the major thylakoid polypeptides in Chlamydomonas reinhardtii y-1 at 25°C and 38°C. J Cell Biol 95: 552–558 (1982).
Hoober JK, Millington RH, D'Angelo LP: Structural similarities between the major polypeptides of thylakoid membranes from Chlamydomonas reinhardtii. Arch Biochem Biophys 202: 221–234 (1980).
Hoober JK, Stegeman WJ: Control of the synthesis of a major polypeptide of chloroplast membranes in Chlamydomonas reinhardtii. J Cell Biol 56: 1–12 (1973).
Hoober JK, Stegeman WJ: Kinetics and regulation of synthesis of the major polypeptides of thylakoid membranes in Chlamydomonas reinhardtii y-1 at elevated temperatures. J Cell Biol 70: 326–337 (1976).
Johanningmeier U, Howell SH: Regulation of light-harvesting chlorophyll-binding protein mRNA accumulation in Chlamydomonas reinhardtii. J Biol Chem 259: 13541–13549 (1984).
Kaufman LS, Briggs WR, Thompson WF: Phytochrome control of specific mRNA levels in developing pea buds. Plant Physiol 78: 388–393 (1985).
Kaufman LS, Thompson WF, Briggs WR: Different red light requirements for phytochrome-induced accumulation of cab RNA and rbcS RNA. Science 226: 1447–1449 (1984).
Kaufman LS, Watson JC, Briggs WR, Thompson WF: Photoregulation of nucleus-encoded transcripts: blue-light regulation of specific transcript abundance. In: Steinbeck KE, Bonitz S, Arntzen CJ, Bogorad L (eds) Molecular Biology of the Photosynthetic Apparatus. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1985) pp 367–372.
Klein O, Senger H: Biosynthetic pathways to δ-aminolevulinic acid induced by blue light in the pigment mutant C-2A′ of Scendesmus obliquus. Photochem Photobiol 27: 203–208 (1978).
Kowallik W: Blue light effects on respiration. Ann Rev Plant Physiol 33: 51–72 (1982).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
Mayfield SP, Taylor WC: Carotenoid-deficient maize seedlings fail to accumulate light-harvesting chlorophyll a/b binding protein (LHCP) mRNA. Eur J Biochem 144: 79–84 (1984).
Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR: Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucl Acids Res 12: 7035–7056 (1984).
Ohad I, Siekevitz P, Palade GE: Biogenesis of chloroplast membranes. I. Plastid dedifferentiation in a dark-grown algal mutant (Chlamydomonas reinhardtii). J Cell Biol 35: 521–552 (1967).
Ohad I, Siekevitz P, Palade GE: Biogenesis of chloroplast membranes. II. Plastid differentiation during greening of a dark-grown algal mutant (Chlamydomonas reinhardtii). J Cell Biol 35: 553–584 (1967).
Oh-hama T, Hase E: Blue light effect on chlorophyll formation in Chlorella protothecoides. Photochem Photobiol 27: 199–202 (1978).
Parthier B: The cooperation of nuclear and plastid genomes in plastid biogenesis and differentiation. Biochem Physiol Pflanzen 177: 283–317 (1982).
Roscher E, Zetsche K: The effects of light quality and intensity on the synthesis of ribulose-1,5-bisphosphate carboxylase and its mRNA in the green alga Chlorogonium elongatum. Planta 167: 582–586 (1986).
Sager R: Photosynthetic pigments in mjtant strains of Chlamydomonas. Carnegie Inst Wash Year Book 60: 374–376 (1961).
Schmidt GW, Bartlett SG, Grossman AR, Cashmore AR, Chua N-H: Biosynthetic pathways of two polypeptide subunits of the light-harvesting chlorophyll a/b complex. J Cell Biol 91: 468–478 (1981).
Schmidt W: Bluelight physiology. BioScience 34: 698–704 (1984).
Schwartzbach SD, Schiff JA, Goldstein NH: Events surrounding the early development of Euglena chloroplasts. Plant Physiol 56: 313–317 (1975).
Senger H: Blue Light Effects in Biological Systems. Springer-Verlag (1984).
Shepherd HS, Ledoigt G, Howell SH: Regulation of light-harvesting chlorophyll-binding protein (LHCP) mRNA accumulation during the cell cycle in Chlamydomonas reinhardtii. Cell 32: 99–107 (1983).
Smith H: Phytochrome and Photomorphogenesis. McGraw-Hill, London, New York (1975).
Spudich JL, Sager R: Regulation of the Chlamydomonas cell cycle by light and dark. J Cell Biol 85: 136–145 (1980).
Staehelin LA, Arntzen CJ: Regulation of chloroplast membrane function: protein phosphorylation changes the spatial organization of membrane components. J Cell Biol 97: 1327–1337 (1983).
Steinbeck KE, McIntosh L, Arntzen CJ, Bogorad L: Indentification of the triazine receptor protein as a chloroplast gene product. Proc Natl Acad Sci USA 78: 7463–7467 (1981).
Steinbiss HJ, Zetsche K: Light and metabolite regulation of the synthesis of ribulose-1,5-bisphosphate carboxylase/oxygenase and the corresponding mRNAs in the unicellular alga Chlorogonium. Planta 167: 575–581 (1986).
Steinmüller K, Zetsche K: Photo- and metabolite regulation for the synthesis of ribulose bisphosphate carboxylase/oxygenase and the phycobiliproteins in the alga Cyanidium caldarium. Plant Physiol 76: 935–939 (1984).
Surzycki S: Synchronously grown cultures of Chlamydomonas reinhardtii. Methods in Enzymol XXIII: 66–73 (1971).
Thompson RJ, Mosig G: Light and genetic determinants in the control of specific chloroplast transcripts in Chlamydomonas reinhardtii. Plant Physiol 76: 1–6 (1984).
Thompson RJ, Davies JP, Mosig G: “Dark-lethality” of certain Chlamydomonas reinhardtii strains is prevented by dim blue light. Plant Physiol 79: 903–907 (1985).
Tobin EM, Silverthorne J: Light regulation of gene expression in higher plants. Ann Rev Plant Physiol 36: 569–593 (1985).
Zuber H: Yearly review — structure and function of light-harvesting complexes and their polypeptides. Photochem Photobiol 42: 821–844 (1985).
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Kindle, K.L. Expression of a gene for a light-harvesting chlorophyll a/b-binding protein in Chlamydomonas reinhardtii: effect of light and acetate. Plant Mol Biol 9, 547–563 (1987). https://doi.org/10.1007/BF00020532
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DOI: https://doi.org/10.1007/BF00020532