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. 1988 Apr 1;106(4):1117–1130. doi: 10.1083/jcb.106.4.1117

Characterization of the thermotolerant cell. II. Effects on the intracellular distribution of heat-shock protein 70, intermediate filaments, and small nuclear ribonucleoprotein complexes

PMCID: PMC2115010  PMID: 2966179

Abstract

Here we further characterize a number of properties inherent to the thermotolerant cell. In the preceding paper, we showed that the acquisition of the thermotolerant state (by a prior induction of the heat-shock proteins) renders cells translationally tolerant to a subsequent severe heat-shock treatment and thereby results in faster kinetics of both the synthesis and subsequent repression of the stress proteins. Because of the apparent integral role of the 70-kD stress proteins in the acquisition of tolerance, we compared the intracellular distribution of these proteins in both tolerant and nontolerant cells before and after a severe 45 degrees C/30-min shock. In both HeLa and rat embryo fibroblasts, the synthesis and migration of the major stress- induced 72-kD protein into the nucleolus and its subsequent exit was markedly faster in the tolerant cells as compared with the nontolerant cells. Migration of preexisting 72-kD into the nucleolus was shown to be dependent upon heat-shock treatment and independent of active heat- shock protein synthesis. Using both microinjection and immunological techniques, we observed that the constitutive and abundant 73-kD stress protein similarly showed a redistribution from the cytoplasm and nucleus into the nucleolus as a function of heat-shock treatment. We show also that other lesions that occur in cells after heat shock can be prevented or at least minimized if the cells are first made tolerant. Specifically, the heat-induced collapse of the intermediate filament cytoskeleton did not occur in cells rendered thermotolerant. Similarly, the disruption of intranuclear staining patterns of the small nuclear ribonucleoprotein complexes after heat-shock treatment was less apparent in tolerant cells exposed to a subsequent heat-shock treatment.

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Selected References

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  1. Arrigo A. P. Cellular localization of HSP23 during Drosophila development and following subsequent heat shock. Dev Biol. 1987 Jul;122(1):39–48. doi: 10.1016/0012-1606(87)90330-7. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Collier N. C., Schlesinger M. J. The dynamic state of heat shock proteins in chicken embryo fibroblasts. J Cell Biol. 1986 Oct;103(4):1495–1507. doi: 10.1083/jcb.103.4.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Hall B. G. Yeast thermotolerance does not require protein synthesis. J Bacteriol. 1983 Dec;156(3):1363–1365. doi: 10.1128/jb.156.3.1363-1365.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Henle K. J., Leeper D. B. Interaction of hyperthermia and radiation in CHO cells: recovery kinetics. Radiat Res. 1976 Jun;66(3):505–518. [PubMed] [Google Scholar]
  7. Henle K. J., Leeper D. B. Modification of the heat response and thermotolerance by cycloheximide, hydroxyurea, and lucanthone in CHO cells. Radiat Res. 1982 May;90(2):339–347. [PubMed] [Google Scholar]
  8. Hightower L. E. Cultured animal cells exposed to amino acid analogues or puromycin rapidly synthesize several polypeptides. J Cell Physiol. 1980 Mar;102(3):407–427. doi: 10.1002/jcp.1041020315. [DOI] [PubMed] [Google Scholar]
  9. Hunt C., Morimoto R. I. Conserved features of eukaryotic hsp70 genes revealed by comparison with the nucleotide sequence of human hsp70. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6455–6459. doi: 10.1073/pnas.82.19.6455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Landry J., Chrétien P. Relationship between hyperthermia-induced heat-shock proteins and thermotolerance in Morris hepatoma cells. Can J Biochem Cell Biol. 1983 Jun;61(6):428–437. doi: 10.1139/o83-058. [DOI] [PubMed] [Google Scholar]
  11. Lee Y. J., Dewey W. C. Effect of cycloheximide or puromycin on induction of thermotolerance by sodium arsenite in Chinese hamster ovary cells: involvement of heat shock proteins. J Cell Physiol. 1987 Jul;132(1):41–48. doi: 10.1002/jcp.1041320106. [DOI] [PubMed] [Google Scholar]
  12. Lerner M. R., Steitz J. A. Snurps and scyrps. Cell. 1981 Aug;25(2):298–300. doi: 10.1016/0092-8674(81)90047-7. [DOI] [PubMed] [Google Scholar]
  13. Li G. C., Werb Z. Correlation between synthesis of heat shock proteins and development of thermotolerance in Chinese hamster fibroblasts. Proc Natl Acad Sci U S A. 1982 May;79(10):3218–3222. doi: 10.1073/pnas.79.10.3218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lim L., Hall C., Leung T., Whatley S. The relationship of the rat brain 68 kDa microtubule-associated protein with synaptosomal plasma membranes and with the Drosophila 70 kDa heat-shock protein. Biochem J. 1984 Dec 1;224(2):677–680. doi: 10.1042/bj2240677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lowe D. G., Moran L. A. Molecular cloning and analysis of DNA complementary to three mouse Mr = 68,000 heat shock protein mRNAs. J Biol Chem. 1986 Feb 15;261(5):2102–2112. [PubMed] [Google Scholar]
  16. Napolitano E. W., Pachter J. S., Chin S. S., Liem R. K. beta-Internexin, a ubiquitous intermediate filament-associated protein. J Cell Biol. 1985 Oct;101(4):1323–1331. doi: 10.1083/jcb.101.4.1323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Napolitano E. W., Pachter J. S., Liem R. K. Intracellular distribution of mammalian stress proteins. Effects of cytoskeletal-specific agents. J Biol Chem. 1987 Feb 5;262(4):1493–1504. [PubMed] [Google Scholar]
  18. Nover L., Munsche D., Neumann D., Ohme K., Scharf K. D. Control of ribosome biosynthesis in plant cell cultures under heat-shock conditions. Ribosomal RNA. Eur J Biochem. 1986 Oct 15;160(2):297–304. doi: 10.1111/j.1432-1033.1986.tb09971.x. [DOI] [PubMed] [Google Scholar]
  19. O'Malley K., Mauron A., Barchas J. D., Kedes L. Constitutively expressed rat mRNA encoding a 70-kilodalton heat-shock-like protein. Mol Cell Biol. 1985 Dec;5(12):3476–3483. doi: 10.1128/mcb.5.12.3476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pelham H. R. Hsp70 accelerates the recovery of nucleolar morphology after heat shock. EMBO J. 1984 Dec 20;3(13):3095–3100. doi: 10.1002/j.1460-2075.1984.tb02264.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Spector D. L., Watt R. A., Sullivan N. F. The v- and c-myc oncogene proteins colocalize in situ with small nuclear ribonucleoprotein particles. Oncogene. 1987 Mar;1(1):5–12. [PubMed] [Google Scholar]
  22. Subjeck J. R., Shyy T. T. Stress protein systems of mammalian cells. Am J Physiol. 1986 Jan;250(1 Pt 1):C1–17. doi: 10.1152/ajpcell.1986.250.1.C1. [DOI] [PubMed] [Google Scholar]
  23. Thomas G. P., Welch W. J., Mathews M. B., Feramisco J. R. Molecular and cellular effects of heat-shock and related treatments of mammalian tissue-culture cells. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):985–996. doi: 10.1101/sqb.1982.046.01.092. [DOI] [PubMed] [Google Scholar]
  24. Voellmy R., Ahmed A., Schiller P., Bromley P., Rungger D. Isolation and functional analysis of a human 70,000-dalton heat shock protein gene segment. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4949–4953. doi: 10.1073/pnas.82.15.4949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wang C., Gomer R. H., Lazarides E. Heat shock proteins are methylated in avian and mammalian cells. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3531–3535. doi: 10.1073/pnas.78.6.3531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Weatherbee J. A., Luftig R. B., Weihing R. R. Purification and reconstitution of HeLa cell microtubules. Biochemistry. 1980 Aug 19;19(17):4116–4123. doi: 10.1021/bi00558a033. [DOI] [PubMed] [Google Scholar]
  27. Welch W. J., Feramisco J. R., Blose S. H. The mammalian stress response and the cytoskeleton: alterations in intermediate filaments. Ann N Y Acad Sci. 1985;455:57–67. doi: 10.1111/j.1749-6632.1985.tb50403.x. [DOI] [PubMed] [Google Scholar]
  28. Welch W. J., Feramisco J. R. Nuclear and nucleolar localization of the 72,000-dalton heat shock protein in heat-shocked mammalian cells. J Biol Chem. 1984 Apr 10;259(7):4501–4513. [PubMed] [Google Scholar]
  29. Welch W. J., Feramisco J. R. Purification of the major mammalian heat shock proteins. J Biol Chem. 1982 Dec 25;257(24):14949–14959. [PubMed] [Google Scholar]
  30. Welch W. J., Feramisco J. R. Rapid purification of mammalian 70,000-dalton stress proteins: affinity of the proteins for nucleotides. Mol Cell Biol. 1985 Jun;5(6):1229–1237. doi: 10.1128/mcb.5.6.1229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Welch W. J. Phorbol ester, calcium ionophore, or serum added to quiescent rat embryo fibroblast cells all result in the elevated phosphorylation of two 28,000-dalton mammalian stress proteins. J Biol Chem. 1985 Mar 10;260(5):3058–3062. [PubMed] [Google Scholar]
  32. Welch W. J., Suhan J. P. Cellular and biochemical events in mammalian cells during and after recovery from physiological stress. J Cell Biol. 1986 Nov;103(5):2035–2052. doi: 10.1083/jcb.103.5.2035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Welch W. J., Suhan J. P. Morphological study of the mammalian stress response: characterization of changes in cytoplasmic organelles, cytoskeleton, and nucleoli, and appearance of intranuclear actin filaments in rat fibroblasts after heat-shock treatment. J Cell Biol. 1985 Oct;101(4):1198–1211. doi: 10.1083/jcb.101.4.1198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Widelitz R. B., Magun B. E., Gerner E. W. Effects of cycloheximide on thermotolerance expression, heat shock protein synthesis, and heat shock protein mRNA accumulation in rat fibroblasts. Mol Cell Biol. 1986 Apr;6(4):1088–1094. doi: 10.1128/mcb.6.4.1088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wu B. J., Morimoto R. I. Transcription of the human hsp70 gene is induced by serum stimulation. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6070–6074. doi: 10.1073/pnas.82.18.6070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yost H. J., Lindquist S. RNA splicing is interrupted by heat shock and is rescued by heat shock protein synthesis. Cell. 1986 Apr 25;45(2):185–193. doi: 10.1016/0092-8674(86)90382-x. [DOI] [PubMed] [Google Scholar]

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