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. 1990 Jan;87(1):374–378. doi: 10.1073/pnas.87.1.374

Identification of heat shock protein hsp70 homologues in chloroplasts.

J S Marshall 1, A E DeRocher 1, K Keegstra 1, E Vierling 1
PMCID: PMC53266  PMID: 2296591

Abstract

Cytoplasmic members of the heat shock protein hsp70 family have recently been implicated in the transport of proteins to the endoplasmic reticulum and mitochondria. In addition, other hsp70 homologues have been found in the endoplasmic reticulum and mitochondria and, at least for the endoplasmic reticulum hsp70 homologue, may be involved in the proper folding and assembly of newly transported proteins. Since chloroplasts are an important site of protein transport in plant cells, we were interested in determining whether hsp70 proteins might be located in this organelle. By using immunoblotting techniques and two antibody preparations against hsp70 proteins, we have identified three chloroplastic proteins of approximately 70 kDa that are related to hsp70 proteins. One of these proteins was tightly associated with the outer envelope membrane and was not exposed at the outer surface of the chloroplasts. The other two were soluble proteins located in the stroma. Steady-state levels of the chloroplastic hsp70 homologues did not change after heat stress nor were any additional hsp70 homologues detected in chloroplasts isolated from heat-stressed plants. We discuss the possible functions of these hsp70 homologues in the transport of proteins into and within chloroplasts.

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Selected References

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

  1. Alfano C., McMacken R. Heat shock protein-mediated disassembly of nucleoprotein structures is required for the initiation of bacteriophage lambda DNA replication. J Biol Chem. 1989 Jun 25;264(18):10709–10718. [PubMed] [Google Scholar]
  2. Bochkareva E. S., Lissin N. M., Girshovich A. S. Transient association of newly synthesized unfolded proteins with the heat-shock GroEL protein. Nature. 1988 Nov 17;336(6196):254–257. doi: 10.1038/336254a0. [DOI] [PubMed] [Google Scholar]
  3. Chappell T. G., Welch W. J., Schlossman D. M., Palter K. B., Schlesinger M. J., Rothman J. E. Uncoating ATPase is a member of the 70 kilodalton family of stress proteins. Cell. 1986 Apr 11;45(1):3–13. doi: 10.1016/0092-8674(86)90532-5. [DOI] [PubMed] [Google Scholar]
  4. Cheng M. Y., Hartl F. U., Martin J., Pollock R. A., Kalousek F., Neupert W., Hallberg E. M., Hallberg R. L., Horwich A. L. Mitochondrial heat-shock protein hsp60 is essential for assembly of proteins imported into yeast mitochondria. Nature. 1989 Feb 16;337(6208):620–625. doi: 10.1038/337620a0. [DOI] [PubMed] [Google Scholar]
  5. Chirico W. J., Waters M. G., Blobel G. 70K heat shock related proteins stimulate protein translocation into microsomes. Nature. 1988 Apr 28;332(6167):805–810. doi: 10.1038/332805a0. [DOI] [PubMed] [Google Scholar]
  6. Cline K., Andrews J., Mersey B., Newcomb E. H., Keegstra K. Separation and characterization of inner and outer envelope membranes of pea chloroplasts. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3595–3599. doi: 10.1073/pnas.78.6.3595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cline K., Werner-Washburne M., Andrews J., Keegstra K. Thermolysin is a suitable protease for probing the surface of intact pea chloroplasts. Plant Physiol. 1984 Jul;75(3):675–678. doi: 10.1104/pp.75.3.675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cline K., Werner-Washburne M., Lubben T. H., Keegstra K. Precursors to two nuclear-encoded chloroplast proteins bind to the outer envelope membrane before being imported into chloroplasts. J Biol Chem. 1985 Mar 25;260(6):3691–3696. [PubMed] [Google Scholar]
  9. Craig E. A., Kramer J., Shilling J., Werner-Washburne M., Holmes S., Kosic-Smithers J., Nicolet C. M. SSC1, an essential member of the yeast HSP70 multigene family, encodes a mitochondrial protein. Mol Cell Biol. 1989 Jul;9(7):3000–3008. doi: 10.1128/mcb.9.7.3000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Deshaies R. J., Koch B. D., Werner-Washburne M., Craig E. A., Schekman R. A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides. Nature. 1988 Apr 28;332(6167):800–805. doi: 10.1038/332800a0. [DOI] [PubMed] [Google Scholar]
  11. Flynn G. C., Chappell T. G., Rothman J. E. Peptide binding and release by proteins implicated as catalysts of protein assembly. Science. 1989 Jul 28;245(4916):385–390. doi: 10.1126/science.2756425. [DOI] [PubMed] [Google Scholar]
  12. Guerriero V., Jr, Raynes D. A., Gutierrez J. A. HSP70-related proteins in bovine skeletal muscle. J Cell Physiol. 1989 Sep;140(3):471–477. doi: 10.1002/jcp.1041400310. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Jensenius J. C., Andersen I., Hau J., Crone M., Koch C. Eggs: conveniently packaged antibodies. Methods for purification of yolk IgG. J Immunol Methods. 1981;46(1):63–68. doi: 10.1016/0022-1759(81)90333-1. [DOI] [PubMed] [Google Scholar]
  15. Kassenbrock C. K., Garcia P. D., Walter P., Kelly R. B. Heavy-chain binding protein recognizes aberrant polypeptides translocated in vitro. Nature. 1988 May 5;333(6168):90–93. doi: 10.1038/333090a0. [DOI] [PubMed] [Google Scholar]
  16. Keegstra K., Cline K. Evidence that Envelope and Thylakoid Membranes from Pea Chloroplasts Lack Glycoproteins. Plant Physiol. 1982 Jul;70(1):232–237. doi: 10.1104/pp.70.1.232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lindquist S., Craig E. A. The heat-shock proteins. Annu Rev Genet. 1988;22:631–677. doi: 10.1146/annurev.ge.22.120188.003215. [DOI] [PubMed] [Google Scholar]
  18. Murakami H., Pain D., Blobel G. 70-kD heat shock-related protein is one of at least two distinct cytosolic factors stimulating protein import into mitochondria. J Cell Biol. 1988 Dec;107(6 Pt 1):2051–2057. doi: 10.1083/jcb.107.6.2051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Olsen L. J., Theg S. M., Selman B. R., Keegstra K. ATP is required for the binding of precursor proteins to chloroplasts. J Biol Chem. 1989 Apr 25;264(12):6724–6729. [PubMed] [Google Scholar]
  20. Palter K. B., Watanabe M., Stinson L., Mahowald A. P., Craig E. A. Expression and localization of Drosophila melanogaster hsp70 cognate proteins. Mol Cell Biol. 1986 Apr;6(4):1187–1203. doi: 10.1128/mcb.6.4.1187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Rothman J. E., Kornberg R. D. Cell biology. An unfolding story of protein translocation. Nature. 1986 Jul 17;322(6076):209–210. doi: 10.1038/322209a0. [DOI] [PubMed] [Google Scholar]
  23. Shinozaki K., Ohme M., Tanaka M., Wakasugi T., Hayashida N., Matsubayashi T., Zaita N., Chunwongse J., Obokata J., Yamaguchi-Shinozaki K. The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression. EMBO J. 1986 Sep;5(9):2043–2049. doi: 10.1002/j.1460-2075.1986.tb04464.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Vierling E., Harris L. M., Chen Q. The major low-molecular-weight heat shock protein in chloroplasts shows antigenic conservation among diverse higher plant species. Mol Cell Biol. 1989 Feb;9(2):461–468. doi: 10.1128/mcb.9.2.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Vierling E., Mishkind M. L., Schmidt G. W., Key J. L. Specific heat shock proteins are transported into chloroplasts. Proc Natl Acad Sci U S A. 1986 Jan;83(2):361–365. doi: 10.1073/pnas.83.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zimmermann R., Sagstetter M., Lewis M. J., Pelham H. R. Seventy-kilodalton heat shock proteins and an additional component from reticulocyte lysate stimulate import of M13 procoat protein into microsomes. EMBO J. 1988 Sep;7(9):2875–2880. doi: 10.1002/j.1460-2075.1988.tb03144.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Zylicz M., Ang D., Liberek K., Georgopoulos C. Initiation of lambda DNA replication with purified host- and bacteriophage-encoded proteins: the role of the dnaK, dnaJ and grpE heat shock proteins. EMBO J. 1989 May;8(5):1601–1608. doi: 10.1002/j.1460-2075.1989.tb03544.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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