Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1992 Sep 15;89(18):8696–8700. doi: 10.1073/pnas.89.18.8696

Identification, characterization, and DNA sequence of a functional "double" groES-like chaperonin from chloroplasts of higher plants.

U Bertsch 1, J Soll 1, R Seetharam 1, P V Viitanen 1
PMCID: PMC49987  PMID: 1356267

Abstract

Chloroplasts of higher plants contain a nuclear-encoded protein that is a functional homolog of the Escherichia coli chaperonin 10 (cpn10; also known as groES). In pea (Pisum sativum), chloroplast cpn10 was identified by its ability to (i) assist bacterial chaperonin 60 (cpn60; also known as groEL) in the ATP-dependent refolding of chemically denatured ribulose-1,5-bisphosphate carboxylase and (ii) form a stable complex with bacterial cpn60 in the presence of Mg.ATP. The subunit size of the pea protein is approximately 24 kDa--about twice the size of bacterial cpn10. A cDNA encoding a spinach (Spinacea oleracea) chloroplast cpn10 was isolated, sequenced, and expressed in vitro. The spinach protein is synthesized as a higher molecular mass precursor and has a typical chloroplast transit peptide. Surprisingly, however, attached to the transit peptide is a single protein, comprised of two distinct cpn10 molecules in tandem. Moreover, both halves of this "double" cpn10 are highly conserved at a number of residues that are present in all cpn10s that have been examined. Upon import into chloroplasts the spinach cpn10 precursor is processed to its mature form of approximately 24 kDa. N-terminal amino acid sequence analysis reveals that the mature pea and spinach cpn10 are identical at 13 of 21 residues.

Full text

PDF

Images in this article

8698Image
on p.8698
8699Image
on p.8699

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barraclough R., Ellis R. J. Protein synthesis in chloroplasts. IX. Assembly of newly-synthesized large subunits into ribulose bisphosphate carboxylase in isolated intact pea chloroplasts. Biochim Biophys Acta. 1980 Jun 27;608(1):19–31. doi: 10.1016/0005-2787(80)90129-x. [DOI] [PubMed] [Google Scholar]
  2. Bloom M. V., Milos P., Roy H. Light-dependent assembly of ribulose-1,5-bisphosphate carboxylase. Proc Natl Acad Sci U S A. 1983 Feb;80(4):1013–1017. doi: 10.1073/pnas.80.4.1013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buchner J., Schmidt M., Fuchs M., Jaenicke R., Rudolph R., Schmid F. X., Kiefhaber T. GroE facilitates refolding of citrate synthase by suppressing aggregation. Biochemistry. 1991 Feb 12;30(6):1586–1591. doi: 10.1021/bi00220a020. [DOI] [PubMed] [Google Scholar]
  4. Chandrasekhar G. N., Tilly K., Woolford C., Hendrix R., Georgopoulos C. Purification and properties of the groES morphogenetic protein of Escherichia coli. J Biol Chem. 1986 Sep 15;261(26):12414–12419. [PubMed] [Google Scholar]
  5. Fayet O., Ziegelhoffer T., Georgopoulos C. The groES and groEL heat shock gene products of Escherichia coli are essential for bacterial growth at all temperatures. J Bacteriol. 1989 Mar;171(3):1379–1385. doi: 10.1128/jb.171.3.1379-1385.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gething M. J., Sambrook J. Protein folding in the cell. Nature. 1992 Jan 2;355(6355):33–45. doi: 10.1038/355033a0. [DOI] [PubMed] [Google Scholar]
  7. Goloubinoff P., Christeller J. T., Gatenby A. A., Lorimer G. H. Reconstitution of active dimeric ribulose bisphosphate carboxylase from an unfoleded state depends on two chaperonin proteins and Mg-ATP. Nature. 1989 Dec 21;342(6252):884–889. doi: 10.1038/342884a0. [DOI] [PubMed] [Google Scholar]
  8. Goloubinoff P., Gatenby A. A., Lorimer G. H. GroE heat-shock proteins promote assembly of foreign prokaryotic ribulose bisphosphate carboxylase oligomers in Escherichia coli. Nature. 1989 Jan 5;337(6202):44–47. doi: 10.1038/337044a0. [DOI] [PubMed] [Google Scholar]
  9. Hartman D. J., Hoogenraad N. J., Condron R., Høj P. B. Identification of a mammalian 10-kDa heat shock protein, a mitochondrial chaperonin 10 homologue essential for assisted folding of trimeric ornithine transcarbamoylase in vitro. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3394–3398. doi: 10.1073/pnas.89.8.3394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hemmingsen S. M., Ellis R. J. Purification and properties of ribulosebisphosphate carboxylase large subunit binding protein. Plant Physiol. 1986 Jan;80(1):269–276. doi: 10.1104/pp.80.1.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hemmingsen S. M., Woolford C., van der Vies S. M., Tilly K., Dennis D. T., Georgopoulos C. P., Hendrix R. W., Ellis R. J. Homologous plant and bacterial proteins chaperone oligomeric protein assembly. Nature. 1988 May 26;333(6171):330–334. doi: 10.1038/333330a0. [DOI] [PubMed] [Google Scholar]
  12. Hendrix R. W. Purification and properties of groE, a host protein involved in bacteriophage assembly. J Mol Biol. 1979 Apr 15;129(3):375–392. doi: 10.1016/0022-2836(79)90502-3. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Laminet A. A., Ziegelhoffer T., Georgopoulos C., Plückthun A. The Escherichia coli heat shock proteins GroEL and GroES modulate the folding of the beta-lactamase precursor. EMBO J. 1990 Jul;9(7):2315–2319. doi: 10.1002/j.1460-2075.1990.tb07403.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lissin N. M., Venyaminov SYu, Girshovich A. S. (Mg-ATP)-dependent self-assembly of molecular chaperone GroEL. Nature. 1990 Nov 22;348(6299):339–342. doi: 10.1038/348339a0. [DOI] [PubMed] [Google Scholar]
  17. Lubben T. H., Gatenby A. A., Donaldson G. K., Lorimer G. H., Viitanen P. V. Identification of a groES-like chaperonin in mitochondria that facilitates protein folding. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7683–7687. doi: 10.1073/pnas.87.19.7683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Martel R., Cloney L. P., Pelcher L. E., Hemmingsen S. M. Unique composition of plastid chaperonin-60: alpha and beta polypeptide-encoding genes are highly divergent. Gene. 1990 Oct 15;94(2):181–187. doi: 10.1016/0378-1119(90)90385-5. [DOI] [PubMed] [Google Scholar]
  19. Martin J., Langer T., Boteva R., Schramel A., Horwich A. L., Hartl F. U. Chaperonin-mediated protein folding at the surface of groEL through a 'molten globule'-like intermediate. Nature. 1991 Jul 4;352(6330):36–42. doi: 10.1038/352036a0. [DOI] [PubMed] [Google Scholar]
  20. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  21. McMullin T. W., Hallberg R. L. A highly evolutionarily conserved mitochondrial protein is structurally related to the protein encoded by the Escherichia coli groEL gene. Mol Cell Biol. 1988 Jan;8(1):371–380. doi: 10.1128/mcb.8.1.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mendoza J. A., Rogers E., Lorimer G. H., Horowitz P. M. Chaperonins facilitate the in vitro folding of monomeric mitochondrial rhodanese. J Biol Chem. 1991 Jul 15;266(20):13044–13049. [PubMed] [Google Scholar]
  23. Morrison R. P., Belland R. J., Lyng K., Caldwell H. D. Chlamydial disease pathogenesis. The 57-kD chlamydial hypersensitivity antigen is a stress response protein. J Exp Med. 1989 Oct 1;170(4):1271–1283. doi: 10.1084/jem.170.4.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Reading D. S., Hallberg R. L., Myers A. M. Characterization of the yeast HSP60 gene coding for a mitochondrial assembly factor. Nature. 1989 Feb 16;337(6208):655–659. doi: 10.1038/337655a0. [DOI] [PubMed] [Google Scholar]
  25. Reed J. E., Cline K., Stephens L. C., Bacot K. O., Viitanen P. V. Early events in the import/assembly pathway of an integral thylakoid protein. Eur J Biochem. 1990 Nov 26;194(1):33–42. doi: 10.1111/j.1432-1033.1990.tb19423.x. [DOI] [PubMed] [Google Scholar]
  26. Saibil H., Dong Z., Wood S., auf der Mauer A. Binding of chaperonins. Nature. 1991 Sep 5;353(6339):25–26. doi: 10.1038/353025b0. [DOI] [PubMed] [Google Scholar]
  27. Salomon M., Fischer K., Flügge U. I., Soll J. Sequence analysis and protein import studies of an outer chloroplast envelope polypeptide. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5778–5782. doi: 10.1073/pnas.87.15.5778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tilly K., Georgopoulos C. Evidence that the two Escherichia coli groE morphogenetic gene products interact in vivo. J Bacteriol. 1982 Mar;149(3):1082–1088. doi: 10.1128/jb.149.3.1082-1088.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tilly K., Murialdo H., Georgopoulos C. Identification of a second Escherichia coli groE gene whose product is necessary for bacteriophage morphogenesis. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1629–1633. doi: 10.1073/pnas.78.3.1629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Van Dyk T. K., Gatenby A. A., LaRossa R. A. Demonstration by genetic suppression of interaction of GroE products with many proteins. Nature. 1989 Nov 23;342(6248):451–453. doi: 10.1038/342451a0. [DOI] [PubMed] [Google Scholar]
  32. Viitanen P. V., Donaldson G. K., Lorimer G. H., Lubben T. H., Gatenby A. A. Complex interactions between the chaperonin 60 molecular chaperone and dihydrofolate reductase. Biochemistry. 1991 Oct 8;30(40):9716–9723. doi: 10.1021/bi00104a021. [DOI] [PubMed] [Google Scholar]
  33. Viitanen P. V., Gatenby A. A., Lorimer G. H. Purified chaperonin 60 (groEL) interacts with the nonnative states of a multitude of Escherichia coli proteins. Protein Sci. 1992 Mar;1(3):363–369. doi: 10.1002/pro.5560010308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Viitanen P. V., Lorimer G. H., Seetharam R., Gupta R. S., Oppenheim J., Thomas J. O., Cowan N. J. Mammalian mitochondrial chaperonin 60 functions as a single toroidal ring. J Biol Chem. 1992 Jan 15;267(2):695–698. [PubMed] [Google Scholar]
  35. Viitanen P. V., Lubben T. H., Reed J., Goloubinoff P., O'Keefe D. P., Lorimer G. H. Chaperonin-facilitated refolding of ribulosebisphosphate carboxylase and ATP hydrolysis by chaperonin 60 (groEL) are K+ dependent. Biochemistry. 1990 Jun 19;29(24):5665–5671. doi: 10.1021/bi00476a003. [DOI] [PubMed] [Google Scholar]
  36. Webb R., Reddy K. J., Sherman L. A. Regulation and sequence of the Synechococcus sp. strain PCC 7942 groESL operon, encoding a cyanobacterial chaperonin. J Bacteriol. 1990 Sep;172(9):5079–5088. doi: 10.1128/jb.172.9.5079-5088.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. van der Vies S. M., Viitanen P. V., Gatenby A. A., Lorimer G. H., Jaenicke R. Conformational states of ribulosebisphosphate carboxylase and their interaction with chaperonin 60. Biochemistry. 1992 Apr 14;31(14):3635–3644. doi: 10.1021/bi00129a012. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES