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. 1986 Nov 1;103(5):2035–2052. doi: 10.1083/jcb.103.5.2035

Cellular and biochemical events in mammalian cells during and after recovery from physiological stress

PMCID: PMC2114370  PMID: 3536957

Abstract

We have examined and compared a number of cellular and biochemical events associated with the recovery process of rat fibroblasts placed under stress by different agents. Metabolic pulse-labeling studies of cells recovering from either heat-shock treatment, exposure to sodium arsenite, or exposure to an amino acid analogue of proline, L-azetidine 2-carboxylic acid, revealed interesting differences with respect to the individual stress proteins produced, their kinetics of induction, as well as the decay in their synthesis during the recovery period. In the initial periods of recovery, the major stress-induced 72-kD protein accumulates within the altered nucleoli in close association with the pre-ribosomal-containing granular region. During the later times of recovery from stress, the nucleoli begin to regain a normal morphology, show a corresponding loss of the 72-kD protein, and the majority of the protein now begins to accumulate within the cytoplasm in three distinct locales: the perinuclear region, along the perimeter of the cells, and finally in association with large phase-dense structures. These latter structures appear to consist of large aggregates of phase-dense material with no obvious encapsulating membrane. More interestingly we show, using double-label indirect immunofluorescence analysis, that much of the perinuclear and cell perimeter-distributed 72-kD protein coincides with the distribution of the cytoplasmic ribosomes. We discuss the possible implications of the presence of the 72-kD stress proteins within the pre-ribosomal-containing granular region of the nucleolus as well as its subsequent colocalization with cytoplasmic ribosomes in terms of the translational changes which occur in cells both during and after recovery from physiological stress.

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Selected References

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  1. Amalric F., Simard R., Zalta J. P. Effet de la température supra-optimale sur les ribonucléoprotéines et le RNA nucléolaire. II. Etude biochimique. Exp Cell Res. 1969 Jun;55(3):370–377. doi: 10.1016/0014-4827(69)90571-0. [DOI] [PubMed] [Google Scholar]
  2. Arrigo A. P., Darlix J. L., Khandjian E. W., Simon M., Spahr P. F. Characterization of the prosome from Drosophila and its similarity to the cytoplasmic structures formed by the low molecular weight heat-shock proteins. EMBO J. 1985 Feb;4(2):399–406. doi: 10.1002/j.1460-2075.1985.tb03642.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ashburner M., Bonner J. J. The induction of gene activity in drosophilia by heat shock. Cell. 1979 Jun;17(2):241–254. doi: 10.1016/0092-8674(79)90150-8. [DOI] [PubMed] [Google Scholar]
  4. Bernstein R. M., Bunn C. C., Hughes G. R., Francoeur A. M., Mathews M. B. Cellular protein and RNA antigens in autoimmune disease. Mol Biol Med. 1984 Apr;2(2):105–120. [PubMed] [Google Scholar]
  5. Catelli M. G., Binart N., Jung-Testas I., Renoir J. M., Baulieu E. E., Feramisco J. R., Welch W. J. The common 90-kd protein component of non-transformed '8S' steroid receptors is a heat-shock protein. EMBO J. 1985 Dec 1;4(12):3131–3135. doi: 10.1002/j.1460-2075.1985.tb04055.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Craig E. A. The heat shock response. CRC Crit Rev Biochem. 1985;18(3):239–280. doi: 10.3109/10409238509085135. [DOI] [PubMed] [Google Scholar]
  7. Francoeur A. M., Peebles C. L., Heckman K. J., Lee J. C., Tan E. M. Identification of ribosomal protein autoantigens. J Immunol. 1985 Oct;135(4):2378–2384. [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. Ingolia T. D., Craig E. A. Four small Drosophila heat shock proteins are related to each other and to mammalian alpha-crystallin. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2360–2364. doi: 10.1073/pnas.79.7.2360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Levinson W., Oppermann H., Jackson J. Transition series metals and sulfhydryl reagents induce the synthesis of four proteins in eukaryotic cells. Biochim Biophys Acta. 1980;606(1):170–180. doi: 10.1016/0005-2787(80)90108-2. [DOI] [PubMed] [Google Scholar]
  12. Lewis M. J., Pelham H. R. Involvement of ATP in the nuclear and nucleolar functions of the 70 kd heat shock protein. EMBO J. 1985 Dec 1;4(12):3137–3143. doi: 10.1002/j.1460-2075.1985.tb04056.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Miyachi K., Tan E. M. Antibodies reacting with ribosomal ribonucleoprotein in connective tissue diseases. Arthritis Rheum. 1979 Jan;22(1):87–93. doi: 10.1002/art.1780220114. [DOI] [PubMed] [Google Scholar]
  15. Nover L., Scharf K. D., Neumann D. Formation of cytoplasmic heat shock granules in tomato cell cultures and leaves. Mol Cell Biol. 1983 Sep;3(9):1648–1655. doi: 10.1128/mcb.3.9.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. Riehl R. M., Sullivan W. P., Vroman B. T., Bauer V. J., Pearson G. R., Toft D. O. Immunological evidence that the nonhormone binding component of avian steroid receptors exists in a wide range of tissues and species. Biochemistry. 1985 Nov 5;24(23):6586–6591. doi: 10.1021/bi00344a042. [DOI] [PubMed] [Google Scholar]
  19. Rubin G. M., Hogness D. S. Effect of heat shock on the synthesis of low molecular weight RNAs in drosophilia: accumulation of a novel form of 5S RNA. Cell. 1975 Oct;6(2):207–213. doi: 10.1016/0092-8674(75)90011-2. [DOI] [PubMed] [Google Scholar]
  20. Sanchez E. R., Toft D. O., Schlesinger M. J., Pratt W. B. Evidence that the 90-kDa phosphoprotein associated with the untransformed L-cell glucocorticoid receptor is a murine heat shock protein. J Biol Chem. 1985 Oct 15;260(23):12398–12401. [PubMed] [Google Scholar]
  21. Shiu R. P., Pouyssegur J., Pastan I. Glucose depletion accounts for the induction of two transformation-sensitive membrane proteinsin Rous sarcoma virus-transformed chick embryo fibroblasts. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3840–3844. doi: 10.1073/pnas.74.9.3840. [DOI] [PMC free article] [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. 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]
  26. 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]
  27. 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]
  28. Welch W. J., Garrels J. I., Thomas G. P., Lin J. J., Feramisco J. R. Biochemical characterization of the mammalian stress proteins and identification of two stress proteins as glucose- and Ca2+-ionophore-regulated proteins. J Biol Chem. 1983 Jun 10;258(11):7102–7111. [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. Whelan S. A., Hightower L. E. Induction of stress proteins in chicken embryo cells by low-level zinc contamination in amino acid-free media. J Cell Physiol. 1985 Feb;122(2):205–209. doi: 10.1002/jcp.1041220207. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Wu F. S., Park Y. C., Roufa D., Martonosi A. Selective stimulation of the synthesis of an 80,000-dalton protein by calcium ionophores. J Biol Chem. 1981 Jun 10;256(11):5309–5312. [PubMed] [Google Scholar]
  34. 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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