Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Sep 2;134(6):1375–1386. doi: 10.1083/jcb.134.6.1375

The molecular chaperone Hsp78 confers compartment-specific thermotolerance to mitochondria

PMCID: PMC2120990  PMID: 8830768

Abstract

Hsp78, a member of the family of Clp/Hsp100 proteins, exerts chaperone functions in mitochondria of S. cerevisiae which overlap with those of mitochondrial Hsp70. In the present study, the role of Hsp78 under extreme stress was analyzed. Whereas deletion of HSP78 does not affect cell growth at temperatures up to 39 decrees C and cellular thermotolerance at 50 degrees C, Hsp78 is crucial for maintenance of respiratory competence and for mitochondrial genome integrity under severe temperature stress (mitochondrial thermotolerance). Mitochondrial protein synthesis is identified as a thermosensitive process. Reactivation of mitochondrial protein synthesis after heat stress depends on the presence of Hsp78, though Hsp78 does not confer protection against heat-inactivation to this process. Hsp78 appears to act in concert with other mitochondrial chaperone proteins since a conditioning pretreatment of the cells to induce the cellular heat shock response is required to maintain mitochondrial functions under severe temperature stress. When expressed in the cytosol, Hsp78 can substitute for the homologous heat shock protein Hsp104 in mediating cellular thermotolerance, suggesting a conserved mode of action of the two proteins. Thus, proteins of the Clp/Hsp100-family located in the cytosol and within mitochondria confer compartment-specific protection against heat damage to the cell.

Full Text

The Full Text of this article is available as a PDF (1.7 MB).

Selected References

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

  1. Beck S. C., De Maio A. Stabilization of protein synthesis in thermotolerant cells during heat shock. Association of heat shock protein-72 with ribosomal subunits of polysomes. J Biol Chem. 1994 Aug 26;269(34):21803–21811. [PubMed] [Google Scholar]
  2. Caplan A. J., Douglas M. G. Characterization of YDJ1: a yeast homologue of the bacterial dnaJ protein. J Cell Biol. 1991 Aug;114(4):609–621. doi: 10.1083/jcb.114.4.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Douglas M. G., Butow R. A. Variant forms of mitochondrial translation products in yeast: evidence for location of determinants on mitochondrial DNA. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1083–1086. doi: 10.1073/pnas.73.4.1083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gambill B. D., Voos W., Kang P. J., Miao B., Langer T., Craig E. A., Pfanner N. A dual role for mitochondrial heat shock protein 70 in membrane translocation of preproteins. J Cell Biol. 1993 Oct;123(1):109–117. doi: 10.1083/jcb.123.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gerner E. W., Schneider M. J. Induced thermal resistance in HeLa cells. Nature. 1975 Aug 7;256(5517):500–502. doi: 10.1038/256500a0. [DOI] [PubMed] [Google Scholar]
  6. Gottesman S., Clark W. P., de Crecy-Lagard V., Maurizi M. R. ClpX, an alternative subunit for the ATP-dependent Clp protease of Escherichia coli. Sequence and in vivo activities. J Biol Chem. 1993 Oct 25;268(30):22618–22626. [PubMed] [Google Scholar]
  7. Gottesman S., Squires C., Pichersky E., Carrington M., Hobbs M., Mattick J. S., Dalrymple B., Kuramitsu H., Shiroza T., Foster T. Conservation of the regulatory subunit for the Clp ATP-dependent protease in prokaryotes and eukaryotes. Proc Natl Acad Sci U S A. 1990 May;87(9):3513–3517. doi: 10.1073/pnas.87.9.3513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hallberg E. M., Hallberg R. L. Translational thermotolerance in Saccharomyces cerevisiae. Cell Stress Chaperones. 1996 Apr;1(1):70–77. doi: 10.1379/1466-1268(1996)001<0070:ttisc>2.3.co;2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hendrick J. P., Hartl F. U. Molecular chaperone functions of heat-shock proteins. Annu Rev Biochem. 1993;62:349–384. doi: 10.1146/annurev.bi.62.070193.002025. [DOI] [PubMed] [Google Scholar]
  10. Horwich A. L. Molecular chaperones. Resurrection or destruction? Curr Biol. 1995 May 1;5(5):455–458. doi: 10.1016/s0960-9822(95)00089-3. [DOI] [PubMed] [Google Scholar]
  11. Hwang B. J., Woo K. M., Goldberg A. L., Chung C. H. Protease Ti, a new ATP-dependent protease in Escherichia coli, contains protein-activated ATPase and proteolytic functions in distinct subunits. J Biol Chem. 1988 Jun 25;263(18):8727–8734. [PubMed] [Google Scholar]
  12. Jämsä E., Vakula N., Arffman A., Kilpeläinen I., Makarow M. In vivo reactivation of heat-denatured protein in the endoplasmic reticulum of yeast. EMBO J. 1995 Dec 1;14(23):6028–6033. doi: 10.1002/j.1460-2075.1995.tb00291.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Katayama Y., Gottesman S., Pumphrey J., Rudikoff S., Clark W. P., Maurizi M. R. The two-component, ATP-dependent Clp protease of Escherichia coli. Purification, cloning, and mutational analysis of the ATP-binding component. J Biol Chem. 1988 Oct 15;263(29):15226–15236. [PubMed] [Google Scholar]
  14. Langer T., Pajic A., Wagner I., Neupert W. Proteolytic breakdown of membrane-associated polypeptides in mitochondria of Saccharomyces cerevisiae. Methods Enzymol. 1995;260:495–503. doi: 10.1016/0076-6879(95)60161-9. [DOI] [PubMed] [Google Scholar]
  15. Lavoie J. N., Gingras-Breton G., Tanguay R. M., Landry J. Induction of Chinese hamster HSP27 gene expression in mouse cells confers resistance to heat shock. HSP27 stabilization of the microfilament organization. J Biol Chem. 1993 Feb 15;268(5):3420–3429. [PubMed] [Google Scholar]
  16. Lee Y. R., Nagao R. T., Key J. L. A soybean 101-kD heat shock protein complements a yeast HSP104 deletion mutant in acquiring thermotolerance. Plant Cell. 1994 Dec;6(12):1889–1897. doi: 10.1105/tpc.6.12.1889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lenne C., Block M. A., Garin J., Douce R. Sequence and expression of the mRNA encoding HSP22, the mitochondrial small heat-shock protein in pea leaves. Biochem J. 1995 Nov 1;311(Pt 3):805–813. doi: 10.1042/bj3110805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Leonhardt S. A., Fearson K., Danese P. N., Mason T. L. HSP78 encodes a yeast mitochondrial heat shock protein in the Clp family of ATP-dependent proteases. Mol Cell Biol. 1993 Oct;13(10):6304–6313. doi: 10.1128/mcb.13.10.6304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Martin J., Horwich A. L., Hartl F. U. Prevention of protein denaturation under heat stress by the chaperonin Hsp60. Science. 1992 Nov 6;258(5084):995–998. doi: 10.1126/science.1359644. [DOI] [PubMed] [Google Scholar]
  20. Mizzen L. A., Welch W. J. Characterization of the thermotolerant cell. I. Effects on protein synthesis activity and the regulation of heat-shock protein 70 expression. J Cell Biol. 1988 Apr;106(4):1105–1116. doi: 10.1083/jcb.106.4.1105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Moczko M., Schönfisch B., Voos W., Pfanner N., Rassow J. The mitochondrial ClpB homolog Hsp78 cooperates with matrix Hsp70 in maintenance of mitochondrial function. J Mol Biol. 1995 Dec 8;254(4):538–543. doi: 10.1006/jmbi.1995.0636. [DOI] [PubMed] [Google Scholar]
  22. Myers A. M., Pape L. K., Tzagoloff A. Mitochondrial protein synthesis is required for maintenance of intact mitochondrial genomes in Saccharomyces cerevisiae. EMBO J. 1985 Aug;4(8):2087–2092. doi: 10.1002/j.1460-2075.1985.tb03896.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Parsell D. A., Kowal A. S., Singer M. A., Lindquist S. Protein disaggregation mediated by heat-shock protein Hsp104. Nature. 1994 Dec 1;372(6505):475–478. doi: 10.1038/372475a0. [DOI] [PubMed] [Google Scholar]
  24. Parsell D. A., Sanchez Y., Stitzel J. D., Lindquist S. Hsp104 is a highly conserved protein with two essential nucleotide-binding sites. Nature. 1991 Sep 19;353(6341):270–273. doi: 10.1038/353270a0. [DOI] [PubMed] [Google Scholar]
  25. Plesofsky-Vig N., Brambl R. Disruption of the gene for hsp30, an alpha-crystallin-related heat shock protein of Neurospora crassa, causes defects in thermotolerance. Proc Natl Acad Sci U S A. 1995 May 23;92(11):5032–5036. doi: 10.1073/pnas.92.11.5032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Praekelt U. M., Meacock P. A. HSP12, a new small heat shock gene of Saccharomyces cerevisiae: analysis of structure, regulation and function. Mol Gen Genet. 1990 Aug;223(1):97–106. doi: 10.1007/BF00315801. [DOI] [PubMed] [Google Scholar]
  27. Prip-Buus C., Westerman B., Schmitt M., Langer T., Neupert W., Schwarz E. Role of the mitochondrial DnaJ homologue, Mdj1p, in the prevention of heat-induced protein aggregation. FEBS Lett. 1996 Feb 12;380(1-2):142–146. doi: 10.1016/0014-5793(96)00049-x. [DOI] [PubMed] [Google Scholar]
  28. Rowley N., Prip-Buus C., Westermann B., Brown C., Schwarz E., Barrell B., Neupert W. Mdj1p, a novel chaperone of the DnaJ family, is involved in mitochondrial biogenesis and protein folding. Cell. 1994 Apr 22;77(2):249–259. doi: 10.1016/0092-8674(94)90317-4. [DOI] [PubMed] [Google Scholar]
  29. Sanchez Y., Lindquist S. L. HSP104 required for induced thermotolerance. Science. 1990 Jun 1;248(4959):1112–1115. doi: 10.1126/science.2188365. [DOI] [PubMed] [Google Scholar]
  30. Sanchez Y., Parsell D. A., Taulien J., Vogel J. L., Craig E. A., Lindquist S. Genetic evidence for a functional relationship between Hsp104 and Hsp70. J Bacteriol. 1993 Oct;175(20):6484–6491. doi: 10.1128/jb.175.20.6484-6491.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sanchez Y., Taulien J., Borkovich K. A., Lindquist S. Hsp104 is required for tolerance to many forms of stress. EMBO J. 1992 Jun;11(6):2357–2364. doi: 10.1002/j.1460-2075.1992.tb05295.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schirmer E. C., Lindquist S., Vierling E. An Arabidopsis heat shock protein complements a thermotolerance defect in yeast. Plant Cell. 1994 Dec;6(12):1899–1909. doi: 10.1105/tpc.6.12.1899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schmitt M., Neupert W., Langer T. Hsp78, a Clp homologue within mitochondria, can substitute for chaperone functions of mt-hsp70. EMBO J. 1995 Jul 17;14(14):3434–3444. doi: 10.1002/j.1460-2075.1995.tb07349.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schröder H., Langer T., Hartl F. U., Bukau B. DnaK, DnaJ and GrpE form a cellular chaperone machinery capable of repairing heat-induced protein damage. EMBO J. 1993 Nov;12(11):4137–4144. doi: 10.1002/j.1460-2075.1993.tb06097.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Skowyra D., Georgopoulos C., Zylicz M. The E. coli dnaK gene product, the hsp70 homolog, can reactivate heat-inactivated RNA polymerase in an ATP hydrolysis-dependent manner. Cell. 1990 Sep 7;62(5):939–944. doi: 10.1016/0092-8674(90)90268-j. [DOI] [PubMed] [Google Scholar]
  36. Solomon J. M., Rossi J. M., Golic K., McGarry T., Lindquist S. Changes in hsp70 alter thermotolerance and heat-shock regulation in Drosophila. New Biol. 1991 Nov;3(11):1106–1120. [PubMed] [Google Scholar]
  37. Squires C. L., Pedersen S., Ross B. M., Squires C. ClpB is the Escherichia coli heat shock protein F84.1. J Bacteriol. 1991 Jul;173(14):4254–4262. doi: 10.1128/jb.173.14.4254-4262.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Squires C., Squires C. L. The Clp proteins: proteolysis regulators or molecular chaperones? J Bacteriol. 1992 Feb;174(4):1081–1085. doi: 10.1128/jb.174.4.1081-1085.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wawrzynow A., Wojtkowiak D., Marszalek J., Banecki B., Jonsen M., Graves B., Georgopoulos C., Zylicz M. The ClpX heat-shock protein of Escherichia coli, the ATP-dependent substrate specificity component of the ClpP-ClpX protease, is a novel molecular chaperone. EMBO J. 1995 May 1;14(9):1867–1877. doi: 10.1002/j.1460-2075.1995.tb07179.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Werner-Washburne M., Stone D. E., Craig E. A. Complex interactions among members of an essential subfamily of hsp70 genes in Saccharomyces cerevisiae. Mol Cell Biol. 1987 Jul;7(7):2568–2577. doi: 10.1128/mcb.7.7.2568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wickner S., Gottesman S., Skowyra D., Hoskins J., McKenney K., Maurizi M. R. A molecular chaperone, ClpA, functions like DnaK and DnaJ. Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):12218–12222. doi: 10.1073/pnas.91.25.12218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wojtkowiak D., Georgopoulos C., Zylicz M. Isolation and characterization of ClpX, a new ATP-dependent specificity component of the Clp protease of Escherichia coli. J Biol Chem. 1993 Oct 25;268(30):22609–22617. [PubMed] [Google Scholar]
  43. Woo K. M., Kim K. I., Goldberg A. L., Ha D. B., Chung C. H. The heat-shock protein ClpB in Escherichia coli is a protein-activated ATPase. J Biol Chem. 1992 Oct 5;267(28):20429–20434. [PubMed] [Google Scholar]
  44. Yaffe M. P., Schatz G. Two nuclear mutations that block mitochondrial protein import in yeast. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4819–4823. doi: 10.1073/pnas.81.15.4819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. van Heusden G. P., Wenzel T. J., Lagendijk E. L., de Steensma H. Y., van den Berg J. A. Characterization of the yeast BMH1 gene encoding a putative protein homologous to mammalian protein kinase II activators and protein kinase C inhibitors. FEBS Lett. 1992 May 11;302(2):145–150. doi: 10.1016/0014-5793(92)80426-h. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES