Skip to main content
SpringerLink
Log in

Comparison of the expression of a plastidic chaperonin 60 in different plant tissues and under photosynthetic and non-photosynthetic conditions

  • Published:
Planta Aims and scope Submit manuscript

Abstract

A partial cDNA which codes for the β-subunit of a plastidic chaperonin 60 (cpn60-β) from rye (Secale cereale L.) leaves was identified and sequenced, except for 46 amino acids of the N-terminus of the mature protein and the transit sequence. This is the first cpn60-β sequence determined for a monocotyledonous plant. Specific antibodies against cpn60-β were affinity-purified from an antiserum raised against the total soluble protein fraction of ribosome-deficient plastids. The localization of cpn60-β in chloroplasts or non-green plastids was confirmed by immunodetection in Percoll gradient-purified organelles. The expression and occurrence of cpn60-β was analysed by immunoblotting with the specific antibodies and Northern hybridization. The cpn60-β protein was constitutively expressed in various green and non-green tissues. It was evenly distributed along the major part of a rye leaf, while highest transcript levels occurred in the youngest and oldest leaf sections. The expression of the cpn60-β protein was not enhanced by a heat-shock treatment at 42 °C. The cpn60-β transcript and protein were more strongly expressed in various non-green, for instance etiolated, 70S-ribosome-deficient 32 °C-grown, or herbicide-bleached tissues, than in green leaves of rye. A rapid increase in the cpn60-β transcript level was also observed when green leaves were transferred from light to darkness while the protein level was not affected. The dark-induced increase in the cpn60-β transcript was totally suppressed in the presence of 2% sucrose. Inhibitor treatments suggested that the change in cpn60-β transcript level was not related to changes of the ATP supply of the tissue. While the large subunit of the photosynthetic protein ribulose-1,5-bisphosphate carboxylase was largely degraded during ripening of tomato fruits, high levels of cpn60-β were detected in tomato chromoplasts and in the yellow flower petals of Narcissus. Low levels of cpn60-β were detected in root tissue.

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

cpn60:

chaperonin 60

HSP:

heat shock protein

Rubisco:

ribulose-1,5-bisphosphate carboxylase/oxygenase

References

  • Baneyx F, Bertsch U, Kalbach CE, van der Vies SM, Soll J, Gatenby AA (1995) Spinach chloroplast cpn21 co-chaperonin possesses two functional domains fused together in a toroidal structure and exhibits nucleotide-dependent binding to plastid chaperonin 60. J Biol Chem 270: 10695–10702

    Google Scholar 

  • Berberich T, Feierabend J (1994) The onset of 70S chloroplast ribosome formation is determined by an early heat-sensitive stage in the ontogeny of rye leaves. Plant Cell Physiol 35: 907–916

    Google Scholar 

  • Bertsch U, Soll J, Seetharam R, Viitanen PV (1992) Identification, characterization, and DNA sequence of a functional “double” groES-like chaperonin from chloroplasts of higher plants. Proc Natl Acad Sci USA 89: 8696–8700

    Google Scholar 

  • Chen GG, Jagendorf AT (1994) Chloroplast molecular chaperonessisted refolding and reconstitution of an active multisubmit coupling factor CF1 core. Proc Natl Acad Sci USA 91: 11497–11501

    Google Scholar 

  • Cheng MY, Hartl F-U, Martin J, Pollock RA, Kalousek F, Neupert W, Hallberg RL, Horwich AL (1989) Mitochondrial heat-shock protein hsp60 is essential for assembly of proteins imported into yeast mitochondria. Nature 337: 620–625

    Google Scholar 

  • Cloney LP, Wu HB, Hemmingsen SM (1992a) Expression of plant chaperonin-60 genes in Escherichia coli. J Biol Chem 267: 23327–23332

    Google Scholar 

  • Cloney LP, Bekkaoui DR, Wood MG, Hemmingsen SM (1992b) Assessment of plant chaperonin-60 gene function in Escherichia coli. J Biol Chem 267: 23333–23336

    Google Scholar 

  • Cohen Y, Yalovsky S, Nechushtai R (1995) Integration and assembly of photosynthetic protein complexes in chloroplast thylakoid membranes. Biochim Biophys Acta 1241: 1–30

    Google Scholar 

  • Dahlin C, Cline K (1991) Developmental regulation of the plastid protein import apparatus. Plant Cell 3: 1131–1140

    Google Scholar 

  • Ehmann B, Kranz M, Mummert E, Schäfer E (1993) Two Tcp-1related but highly divergent gene families exist in oat encoding proteins of assumed chaperone function. FEBS Lett 336: 313–316

    Google Scholar 

  • Feierabend J (1982) Inhibition of chloroplast ribosome formation by heat in higher plants. In: Edelman M, Hallick RB, Chua N-H (eds) Methods in chloroplast molecular biology. Elsevier Biomedical Press, pp 671–680

  • Feierabend J, Berberich T (1991) Heat-induced ribosome-deficiency of plastids — mechanism and applications. In: Mache R, Stutz E, Subramanian AR (eds) The translational apparatus of photosynthetic organelles. Springer, Berlin, Heidelberg, New York, pp 215–227

    Google Scholar 

  • Feierabend J, Winkelhüsener T, Kemmerich P, Schulz U (1982) Mechanism of bleaching in leaves treated with chlorosis-inducing herbicides. Z Naturforsch 37c: 898–907

    Google Scholar 

  • Feierabend J, Schlüter W, Tebartz K (1988) Unassembled polypeptides of the plastidic ribosomes in heat-treated 70S ribosome deficient rye leaves. Planta 174: 542–550

    Google Scholar 

  • Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13

    CAS  PubMed  Google Scholar 

  • Gatenby AA, Ellis RJ (1990) Chaperone function: The assembly of ribulose bisphosphate carboxylase-oxygenase. Annu Rev Cell Biol 6: 125–149

    Google Scholar 

  • Gatenby AA, Viitanen PV (1994) Structural and functional aspects of chaperonin-mediated protein folding. Annu Rev Plant Physiol Plant Mol Biol 45: 469–491

    Google Scholar 

  • Gerhardt B, Beevers H (1968) Influence of sucrose on protein determination by the Lowry procedure. Anal Biochem 24: 337–339

    Google Scholar 

  • Gray JC, Row PE (1995) Protein translocation across chloroplast envelope membranes. Trends Cell Biol 5: 243–247

    Google Scholar 

  • Grimm R, Speth V, Gatenby AA, Schäfer E (1991) GroEL-related molecular chaperones are present in the cytosol of oat cells. FEBS Lett 286: 155–158

    Google Scholar 

  • Hartl F-U, Hlodan R, Langer T (1994) Molecular chaperones in protein folding: the art of avoiding sticky situations. Trends Biol Sci 19: 20–25

    Google Scholar 

  • Hemmingsen SM, Wooford CA, van der Vies SM, Tilly K, Dennis DT, Georgopoulos CP, Hendrix RW, Ellis RJ (1988) Homologous plant and bacterial proteins chaperone oligomeric protein assembly. Nature 333: 330–334

    Google Scholar 

  • Hendrick JP, Hartl F-U (1993) Molecular chaperone functions of heat-shock proteins. Annu Rev Biochem 62: 349–84

    Google Scholar 

  • Höinghaus R, Feierabend J (1983) Rapid purification of intact chloroplasts and heat-bleached ribosome-deficient plastids from rye leaves on discontinuous Percoll gradients. Protoplasma 118: 114–120

    Google Scholar 

  • Jakob U, Buchner J (1994) Assisting spontaneity: the role of HSP90 and small HSPs as molecular chaperones. Trends Biol Sci 19: 205–211

    Google Scholar 

  • Jang J-C, Sheen J (1994) Sugar sensing in higher plants. Plant Cell 6: 1665–1679

    Google Scholar 

  • Kim S-R, Costa MA, An G (1991) Sugar response element enhances wound response of potato proteinase inhibitor II promoter in transgenic tobacco. Plant Mol Biol 17: 973–983

    Google Scholar 

  • Koning AJ, Rose R, Comai L (1992) Developmental expression of tomato heat-shock cognate protein 80. Plant Physiol 100: 801–811

    Google Scholar 

  • Krishna P, Sacco M, Cherutti JF, Hill S (1995) Cold-induced accumulation of hsp90 transcripts in Brassica napus. Plant Physiol 107: 915–923

    Google Scholar 

  • Kruse E, Liu Z, Kloppstech K (1993) Expression of heat shock proteins during development of barley. Plant Mol Biol 23: 111–122

    Google Scholar 

  • Kyhse-Andersen J (1984) Electroblotting of multiple gels: A simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods 10: 203–209

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    PubMed  Google Scholar 

  • Lam H-M, Peng S-Y, Coruzzi GM (1994) Metabolic regulation of the gene encoding glutamine-dependent asparagine synthetase in Arabidopsis thaliana. Plant Physiol 106: 1347–1357

    Google Scholar 

  • Logemann J, Schell J, Willmitzer L (1987) Improved method for the isolation of RNA from plant tissues. Anal Biochem 163: 16–20

    CAS  PubMed  Google Scholar 

  • Lubben TH, Donaldson GK, Viitanen PV, Gatenby AA (1989) Several proteins imported into chloroplasts form stable complexes with the GroEL-related chloroplast molecular chaperone. Plant Cell 1: 1223–1230

    Google Scholar 

  • Marshall JS, Keegstra K (1992) Isolation and characterization of a cDNA clone encoding the major HSP70 of the pea chloroplastic stroma. Plant Physiol 100: 1048–1054

    Google Scholar 

  • Marshall JS, DeRocher AE, Keegstra K, Vierling E (1990) Identification of heat shock protein hsp70 homologues in chloroplasts. Proc Natl Acad Sci USA 87: 374–378

    Google Scholar 

  • Martel R, Cloney LP, Pelcher LE, Hemmingsen SM (1990) Unique composition of plastid chaperonin-60: α and β polypeptide encoding genes are highly divergent. Gene 94: 181–187

    Google Scholar 

  • Mori M, Murata K, Kubota H, Yamamoto A, Matsushiro A, Morita T (1992) Cloning of a cDNA encoding the Tcp-1 (t complex polypeptide 1) homologue of Arabidopsis thaliana. Gene 122: 381–382

    Google Scholar 

  • Nover L (1991) Heat shock response. CRC Press, Boca Raton, Ann Arbor, Boston, London

    Google Scholar 

  • Otto S, Feierabend J (1989) Enzymes of starch and sugar phosphate metabolism in achlorophyllous ribosome-deficient plastids from high-temperature-grown rye leaves. Physiol Plant 76: 65–73

    Google Scholar 

  • Prasad TK, Stewart CR (1992) cDNA clones encoding Arabidopsis thaliana and Zea mays mitochondrial chaperonin HSP60 and gene expression during seed germination and heat shock. Plant Mol Biol 18: 873–885

    Google Scholar 

  • Reiß T, Bergfeld R, Link G, Thien W, Mohr H (1983) Photooxidative destruction of chloroplasts and its consequences for cytosolic enzyme levels and plant development. Planta 159: 518–528

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual. Cold Spring Harbor Lab Press, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467

    CAS  PubMed  Google Scholar 

  • Schmidt M, Svendsen I, Feierabend J (1995) Analysis of the primary structure of the chloroplast isozyme of triosephosphate isomerase from rye leaves by protein and cDNA sequencing indicates a eukaryotic origin of its gene. Biochim Biophys Acta 1261: 257–264

    Google Scholar 

  • Schmitz G, Schmidt M, Feierabend J (1996) Characterization of a plastid-specific HSP90 homologue: Identification of a cDNA sequence, phylogenetic descendence and analysis of its mRNA and protein expression. Plant Mol Biol 30: 479–492

    Google Scholar 

  • Takahashi T, Naito S, Komeda Y (1991) Isolation and analysis of the expression of two genes for the 81 kilodalton heat-shock proteins from Arabidopsis. Plant Physiol 99: 383–390

    Google Scholar 

  • Thompson MD, Paavola CD, Lenvik TR, Gantt JS (1995) Chlamydomonas transcripts encoding three divergent plastid chaperonins are heat-inducible. Plant Mol Biol 27: 1031–1035

    Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354

    CAS  PubMed  Google Scholar 

  • Tseng T-C, Tsai T-H, Lue M-Y, Lee H (1995) Identification of sucrose-regulated genes in cultured rice cells using mRNA differential display. Gene 161: 179–182

    Google Scholar 

  • Tsugeki R, Mori H, Nishimura M (1992) Purification, cDNA cloning and Northern-blot analysis of mitochondrial chaperonin 60 from pumpkin cotyledons. Eur J Biochem 209: 453–458

    Google Scholar 

  • Vierling E (1991) The roles of heat shock proteins in plants. Annu Rev Plant Physiol Plant Mol Biol 42: 579–620

    Google Scholar 

  • Waegemann K, Soll J (1991) Characterization of the protein import apparatus in isolated outer envelopes of chloroplasts. Plant J 1: 149–158

    Google Scholar 

  • Winter U, Feierabend J (1990) Multiple coordinate controls contribute to a balanced expression of ribulose-1,5-bisphosphate carboxylase/oxygenase subunits in rye leaves. Eur J Biochem 187: 445–453

    Google Scholar 

  • Yost HJ, Lindquist S (1986) RNA splicing is interrupted by heat shock and is rescued by heat shock protein synthesis. Cell 45: 185–193

    Google Scholar 

  • Zabaleta E, Oropeza A, Jiménez B, Salerno G, Crespi M, Herrera-Estrella L (1992) Isolation and characterization of genes encoding chaperonin 60β from Arabidopsis thaliana. Gene 111: 175–181

    Google Scholar 

  • Zabaleta E, Oropeza A, Assad N, Mandel A, Salerno G, Herrera-Estrella L (1994) Antisense expression of chaperonin 60β in transgenic tobacco plants leads to abnormal phenotypes and altered distribution of photoassimilates. Plant J 6: 425–432

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Botanisches Institut, J.W. Goethe-Universität, Postfach 11 1932, D-60054, Frankfurt am Main, Germany

    G. Schmitz, M. Schmidt & J. Feierabend

Authors
  1. G. Schmitz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Schmidt
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. Feierabend
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to J. Feierabend.

Additional information

Financial support by the Fonds der Chemischen Industrie, Germany, is greatly appreciated.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schmitz, G., Schmidt, M. & Feierabend, J. Comparison of the expression of a plastidic chaperonin 60 in different plant tissues and under photosynthetic and non-photosynthetic conditions. Planta 200, 326–334 (1996). https://doi.org/10.1007/BF00200300

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00200300

Key words

Search

Navigation