Skip to main content
SpringerLink
Log in

Rubisco, rubisco activase and ribulose-5-phosphate kinase gene expression and polypeptide accumulation in a tobacco mutant defective in chloroplast protein synthesis

  • Regular Papers
  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

Expression of the genes for ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; rbcS and rbcL), Rubisco activase (rca) and ribulose-5-phosphate (Ru5-P) kinase (prk) and accumulation of the polypeptides was examined in chlorophyllous and chlorotic sectors of the DP1 mutant of Nicotiana tabacum. Plastids from chlorotic sectors of this variegated plastome mutant contained 30S and 50S ribosomal subunits, but had abnormally low levels of plastid polysomes. Consequently, mutant plastids were translationally repressed, unable to synthesize plastid-encoded polypeptides including the large subunit of Rubisco despite the presence of the corresponding mRNAs. Transcripts of rbcS accumulated to near wild type levels in chlorotic sectors, but there was little accumulation of the Rubisco small subunit (SS) polypeptide or holoenzyme. Messenger-RNA isolated from chlorotic sectors effectively directed the synthesis of Rubisco SS in vitro suggesting that posttranslational factors were responsible for the decrease in Rubisco SS abundance. Transcripts of rca and prk also accumulated to near wild type levels in chlorotic sectors and a diurnal rhythm in the abundance of rca mRNA was detected in green and chlorotic sectors. Despite the low abundance of Rubisco holoenzyme in chlorotic sectors, Rubisco activase and Ru5-P kinase polypeptides accumulated to significant levels. Activities of Rubisco and Ru5-P kinase paralleled protein levels, indicating that active forms of these enzymes were present in chlorotic sectors. The data indicate that the developmental events governing the accumulation of Rubisco activase and Ru5-P kinase polypeptides and the diurnal regulation of rca expression were not dependent on the attainment of photosynthetically competent plastids or the accumulation of Rubisco.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

LS:

large subunit of Rubisco

Ru5-P:

kinase-ribulose-5-phosphate kinase

Rubisco:

ribulose-1,5-bisphosphate carboxylase/oxygenase

SS:

small subunit of Rubisco

References

  • Andrews TJ and Lorimer GH (1987) Rubisco: Structure, mechanisms, and prospects for improvement. In: Hatch MD and Boardman NK (eds) The Biochemistry of Plants, Vol 10, pp 131–218. Academic Press, New York

    Google Scholar 

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts. Polyphenol-oxidase in Beta vulgaris. Plant Physiol 25: 1–15

    Google Scholar 

  • Barkan A (1993) Nuclear mutants of maize with defects in chloroplast polysome assembly have altered chloroplast RNA metabolism. Plant Cell 5: 389–402

    Google Scholar 

  • Baumgartner BJ, Rapp JC and Mullet JE (1989) Plastid transcription activity and DNA copy number increase early in barley chloroplast development. Plant Physiol 89: 1011–1018

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 73: 248–274

    Google Scholar 

  • Burk LG, Stewart RN and Dermen H (1964) Histogenesis and genetics of a plastid-controlled chlorophyll variegation in tobacco. Am J Bot 51 (7): 713–724

    Google Scholar 

  • Crafts-Brandner SB, Salvucci ME and Egli DB (1990) Changes in ribulosebisphosphate carboxylase/oxygenase and ribulose 5-phosphate kinase abundances and photosynthetic capacity during leaf senescence. Photosynthe Res 23: 223–230

    Google Scholar 

  • Gatenby AA and Ellis RJ (1990) Chaperone function. The assembly of ribulosebisphosphate carboxylase-oxygenase. Ann Rev Cell Biol 6: 125–149

    Google Scholar 

  • Gilmartin PM, Sarokin L, Memelink J and Chua N-H (1990) Molecular light switches for plant genes. Plant Cell 2: 369–378

    Google Scholar 

  • Giuliano G, Hoffman NE, Ko K, Scolnik PA and Cashmore AR (1988) A light-entrained circadian clock controls transcription of several plant genes. EMBO J 7: 3635–3642

    Google Scholar 

  • Hess WR, Muller A, Nagy F and Borner T (1994) Ribosome-deficient plastids affect transcription of light-induced nuclear genes: Genetic evidence for a plastid-derived signal. Mol Gen Genet 242: 305–312

    Google Scholar 

  • Hurwitz J, Weissbach A, Horecker BL and Smyrniotis PZ (1956) Spinach phosphoribulokinase. J Biol Chem 218: 769–783

    Google Scholar 

  • Jiang CZ, Quick WP, Alred R, Kliebenstein D and Rodermel SR (1994) Antisense RNA inhibition of Rubisco activase expression. Plant J 5(6): 787–798

    Google Scholar 

  • Klein RR and Mullet JE (1986) Regulation of chloroplast-encoded chlorophyll-binding protein translation during higher plant chloroplast biogenesis. J Biol Chem 261: 11138–11145

    Google Scholar 

  • Klein RR and Mullet JE (1987) Control of gene expression during higher plant chloroplast biogenesis. Protein synthesis and transcript levels of psbA, psaA-psaB, and rbcL in dark-grown and illuminated barley seedlings. J Biol Chem 262: 4341–4348

    Google Scholar 

  • Klein RR and Mullet JE (1990) Light-induced transcription of chloroplast genes. PsbA transcription is differentially enhanced in illuminated barley. J Biol Chem 265: 1895–1902.

    Google Scholar 

  • Klein RR and Salvucci ME (1992) Photoaffinity labeling of mature and precursor forms of the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase after expression in Escherichia coli. Plant Physiol 98: 546–553

    Google Scholar 

  • Klein RR, Mason HS and Mullet JE (1988) Light-regulated translation of chloroplast proteins. I. Transcripts of psaA-psaB, psbA and rbcL are associated with polysomes in dark-grown and illuminated barley seedlings. J Cell Biol 106: 289–301

    Google Scholar 

  • Klein RR, Crafts-Brandner SJ and Salvucci ME (1993) Cloning and developmental expression of the sucrose-phosphate-synthase gene from spinach. Planta 190: 498–510

    Google Scholar 

  • Kuhlemeier C, Green PJ and Chua N-H (1987) Regulation of gene expression in higher plants. Ann Rev Plant Physiol 38: 221–257

    Google Scholar 

  • Martino-Catt S and Ort DR (1992) Low temperature interrupts circadian regulation of transcriptional activity in chilling-sensitive plants. Proc Natl Acad Sci 89: 3731–3735

    Google Scholar 

  • Mate CJ, Hudson GS, von Caemmerer S, Evans JR and Andrews TJ (1993) Reduction of ribulosebisphosphate carboxylase activase levels in tobacco (Nicotiana tabacum) by antisense RNA reduces ribulose bisphosphate carbamylation and impairs photosynthesis. Plant Physiol 102: 1119–1128

    Google Scholar 

  • Michalowski CB, DeRocher EJ, Bohnert HJ and Salvucci ME (1992) Phosphoribulokinase from ice plant: Transcription, transcripts and protein expression during environmental stress. Photosynth Res 31: 127–138

    Google Scholar 

  • Mullet JE (1988) Chloroplast development and gene expression. Ann Rev Plant Physiol Plant Mol Biol 39: 475–502

    Google Scholar 

  • Mullet JE (1993) Dynamic regulation of chloroplast transcription. Plant Physiol 103: 309–313

    Google Scholar 

  • Mullet JE, Klein RR and Grossman AR (1986) Optimization of protein synthesis in isolated higher plant chloroplasts. Identification of paused translation intermediates. Eur J Biochem 155: 331–338

    Google Scholar 

  • Nagy F, Kay SA and Chua N-H (1988a) Gene regulation by phytochrome. TIG 4: 37–41

    Google Scholar 

  • Nagy F, Kay SA and Chua N-H (1988b) A circadian clock regulates transcription of the wheat Cab-1 gene. Genes Dev 2: 376–382

    Google Scholar 

  • Pilgrim ML and McClung CR (1993) Differential involvement of the circadian clock in the expression of genes required for ribulose-1,5-bisphosphate carboxylase/oxygenase synthesis, assembly, and activation in Arabidopsis thaliana. Plant Physiol 103: 553–564

    Google Scholar 

  • Raines CA, Longstaff M, Lloyd JC and Dyer TA (1989) Complete coding sequence of wheat phosphoribulokinase: Developmental and light-dependent expression of the mRNA. Mol Gen Genet 220: 43–48

    Google Scholar 

  • Raines CA, Lloyd JC and Dyer TA (1991) Molecular biology of the C3 photosynthetic carbon reduction cycle. Photosynth Res 27: 1–14

    Google Scholar 

  • Rochaix J-D (1992) Post-transcriptional steps in the expression of chloroplast genes. Ann Rev Cell Biol 8: 1–28

    Google Scholar 

  • Rodermel SR, Abbott MS and Bogorad L (1988) Nuclear-Organelle interactions: Nuclear antisense gene inhibits ribulose bisphosphate carboxylase enzyme levels in transformed tobacco plants. Cell 55: 673–681

    Google Scholar 

  • Rundle SJ and Zielinski RE (1991a) Organization and expression of two tandemly oriented genes encoding ribulose bisphosphate carboxylase/oxygenase activase in barley. J Biol Chem 266: 4677–4685

    Google Scholar 

  • Rundle SJ and Zielinski RE (1991b) Alterations in barley ribulose-1,5-bisphosphate carboxylase/oxygenase activase gene expression during development and in response to illumination. J Biol Chem 266: 14802–14807

    Google Scholar 

  • Salvucci ME and Klein RR (1994) Site-directed mutagenesis of a reactive lysine residue (Lys-247) of rubisco activase. Arch Biochem Biophys (in press)

  • Salvucci ME, Portis ARJr and Ogren WL (1985) A soluble chloroplast protein catalyzes ribulose bisphosphate carboxylase/oxygenase activation in vivo. Photosynth Res 7: 193–201

    Google Scholar 

  • Salvucci ME, Werneke JM, Ogren WL and PortisJr AR (1987) Purification and species distribution of rubisco activase. Plant Physiol 84: 930–936

    Google Scholar 

  • Salvucci ME, Rajagopalan K, Sievert G, Haley BE Watt DS (1993) Photoaffinity labeling of ribulose-1,5-bisphosphate carboxylase/oxygenase activase with ATP gamma-benzophenone. J Biol Chem 268: 14239–14244

    Google Scholar 

  • Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  • Schmidt GW and Mishkind ML (1983) Rapid degradation of unassembled ribulose 1,5-bisphosphate carboxylase small subunits in chloroplasts. Proc Natl Acad Sci (USA) 80: 2632–2636

    Google Scholar 

  • Shinozaki K, Ohme M, Tanaka M, Wakasugi T, Hayashida N et al. (1986) The complete nucleotide sequence of the tobacco chloroplast genome: Its gene organization and expression. EMBO J 5: 2043–2049

    Google Scholar 

  • Somerville CR (1986) Analysis of photosynthesis with mutants of higher plants and algae. Ann Rev Plant Physiol 37: 467–507

    Google Scholar 

  • Spreitzer RJ, Goldschmidt-Clermont M, Rahire M and Rochaix JD (1985) Non-sense mutations in the Chlamydomonas chloroplast gene that codes for the large subunit of ribulosebisphosphate carboxylase/oxygenase. Proc Natl Acad Sci (USA) 84: 5460–5464

    Google Scholar 

  • Taylor WC (1989a) Regulatory interactions between nuclear and plastid genomes. Ann Rev Plant Physiol Plant Mol Biol 40: 211–233

    Google Scholar 

  • Taylor WC (1989a) Transcriptional regulation by a circadian rhythm. Plant Cell 1: 259–264

    Google Scholar 

  • Watillon B, Kettmann R, Boxus P and Burny A (1993) Developmental and circadian pattern of rubisco activase mRNA accumulation in apple plants. Plant Mol Biol 23: 501–509

    Google Scholar 

  • Zielinski RE, Werneke JM and Jenkins ME (1989) Coordinate expression of rubisco activase and rubisco during barley leaf cell development. Plant Physiol 90: 516–521

    Google Scholar 

Download references

Author information

Author notes

  1. Robert R. Klein

    Present address: US Department of Agriculture, Agricultural Research Service, ERRC, 600 E. Mermaid Lane, 19118, Philadelphia, PA, USA

  2. Michael E. Salvucci

    Present address: Agricultural Research Service, WCRL, US Department of Agriculture, 4135 E. Broadway Road, 85040-8830, Phoenix, AZ, USA

Authors and Affiliations

  1. US Department of Agriculture, Agricultural Research Service and Agronomy Department, University of Kentucky, 40546-0076, Lexington, KY, USA

    Robert R. Klein & Michael E. Salvucci

Authors
  1. Robert R. Klein
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michael E. Salvucci
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Klein, R.R., Salvucci, M.E. Rubisco, rubisco activase and ribulose-5-phosphate kinase gene expression and polypeptide accumulation in a tobacco mutant defective in chloroplast protein synthesis. Photosynth Res 43, 213–223 (1995). https://doi.org/10.1007/BF00029934

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00029934

Key words

Search

Navigation