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Intracellular localization of constitutive and inducible heat shock protein 70 in rat liver after in vivo heat stress

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Abstract

The level and intracellular redistribution of the two nucleo-cytoplasmic members of 70 kDa heat shock protein family (constitutive, Hsc70 or Hsp73, and inducible, Hsp72) were studied in rat liver during a 24-h period after exposure of the animals to 41 °C whole body hyperthermic stress. The examined proteins were detected in the liver cytosol and nuclei by Western blotting and immunohistochemical staining of paraffin sections, as well as by immnocytochemical staining of isolated nuclear smears. All three techniques applied were based on the use of monoclonal antibodies recognizing both constitutive and inducible Hsp70 isoforms or only the inducible isoform, and gave consistent results. The exposure of the animals to in vivo heat stress was shown to induce the synthesis of otherwise non-existing Hsp72, rendering Hsc70 level unchanged in comparison to unstressed controls. However, immediately after the stress the intracellular redistribution of Hsc70, i.e. its nuclear accumulation, was observed. The maximal level of Hsp70 both in the cytoplasm and in the nuclei was registered 5 h after the stress, which coincided with the maximal level of Hsp72 induction. The alterations in the level and intracellular distribution of examined proteins were still noticeable 24 h after the stress. The results of this study could shed some more light on, as yet uncertain, differences between cellular functions of these two proteins, as well as on the role of the constitutive form under normal and stress conditions. (Mol Cell Biochem 265: 27–35, 2004)

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References

  1. Georgopoulos C, Welch WJ: Role of the major heat shock proteins as molecular chaperones. Ann Rev Cell Biol 9: 601–634, 1993

    Google Scholar 

  2. Kiang JG, Tsokos GC: Heat shock protein 70 kDa: Molecular biology, biochemistry and physiology. Pharmacol Ther 80: 183–201, 1998

    Google Scholar 

  3. Schiaffonati L, Tacchini L, Pappalardo C: Heat shock response in the liver: Expression and regulation of the HSP70 gene family and early response genes after in vivo hyperthermia. Hepatology 20: 975–983, 1994

    Google Scholar 

  4. Craig EA, Baxter BK, Becker J, Halladay J, Ziegelhoffer T: Cytosolic Hsp70s of Saccharomyces cerevisiae: Roles in protein synthesis, pro-tein translocation, proteolysis, and regulation. In: R.I. Morimoto, A. Tissieres, C. Geordopoulos (eds.). The Biology of Heat Shock Proteins and Molecular Chaperones. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 1994, pp. 31–53

    Google Scholar 

  5. Olazabal UE, Pfaff DW, Mobbs CV: Sex differences in the regulation of heat shock protein 70 kDa and 90 kDa in the rat ventromedial hy-pothalamus by estrogen. Brain Res 596: 311–314, 1992

    Google Scholar 

  6. Udelsman R, Li D-G, Stagg CA: Adrenergic regulation of adrenal and aortic heat shock protein. Surgery 116: 177–182, 1994

    Google Scholar 

  7. Milarski KL, Morimoto RI: Expression of human hsp70 during the synthetic phase of the cell cycle. Proc Natl Acad Sci USA 83: 9517–9521, 1986

    Google Scholar 

  8. Zakeri ZF, Wolgemuth DJ: Developmental stage-specific expression of the hsp70 gene family during differentiation of the mammalian germ line. Mol Cell Biol 7: 1791–1796, 1987

    Google Scholar 

  9. Raibowol KT, Mizzen LA, Welch WJ: Heat shock is lethal to fibroblasts microinjected with antibodies against Hsp70. Science 242: 433–436, 1988

    Google Scholar 

  10. Beckman RP, Mizzen LA, Welch WJ: Interaction of hsp70 with newly synthetized proteins: Implications for protein folding and assembly. Sci-ence 248: 850–854, 1990

    Google Scholar 

  11. Hightower LE: Heat shock, stress proteins, chaperones and proteotoxicity. Cell 66: 1–20, 1991

    Google Scholar 

  12. Ellis RJ: The molecular chaperon concept. Semin Cell Biol 1: 1–9, 1990

    Google Scholar 

  13. Manzerra P, Rush SJ, Brown JR: Tissue-specific differences in heat shock protein hsc70 and hsp72 in the control and hyperthermic rabbit. J Cell Physiol 170: 130–137, 1997

    Google Scholar 

  14. Thayer JM, Mirkes PE: Induction of Hsp72 and transient nuclear lo-calization of Hsp73 and Hsp72 correlate with the acquisition and loss of thermotolerance in postimplantation rat embyos. Develop Dynamics 208: 227–243, 1997

    Google Scholar 

  15. Foster JA, Brown JR: Intracellular localization of heat shock m/RNAs (hsc70 and hsp70) to neural cell bodies and processes in the control and hyperthermic rabbit brain. J Neurosci Res 46: 652–665, 1996

    Google Scholar 

  16. Dean DO, Kent CR, Tytell M: Constitutive and inducible heat shock protein 70 immunoreactivity in the normal rat eye. Invest Ophthalmol Visual Sci 40: 2952–2962, 1999

    Google Scholar 

  17. Ellis S, Killender M, Anderson RL: Heat-induced alterations in the localization of HSP72 and HSP73 as measured by indirect immuno-histochemistry and immunogold electron microscopy. J Histochem Cytochem 48: 321–331, 2000

    Google Scholar 

  18. Currie RW, White FP: Trauma-induced protein in rat tissues: A physiological role for a heat-shock protein? Science 214: 72–73, 1981

    Google Scholar 

  19. Chauveau J, Moule J, Roulier C: Isolation of pure and unaltered liver nuclei. Morphology and biochemical composition. Exp Cell Res 11: 317–321, 1956

    Google Scholar 

  20. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970

    Google Scholar 

  21. Lowry OH, Rosebrough NJ, Farr AJ, Randall RJ: Protein measure-ment with the Folin phenol reagent. J Biol Chem 123: 265–275, 1951

    Google Scholar 

  22. Burton K: A study of the conditions and mechanisms of the dipheny-lamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J 62: 315–323, 1956

    Google Scholar 

  23. Velazquez JM, Lindquist S: Hsp70: Nuclear concentration during en-vironmental stress, cytoplasmic storage during recovery. Cell 36: 655–663, 1984

    Google Scholar 

  24. Welch WJ, Feramisco JR: Nuclear and nucleolar localization of the 72,000-dalton heat shock protein in heat shocked mammalian cells. J Biol Chem 259: 4501–4513, 1984

    Google Scholar 

  25. Leoni S, Brambilla D, Risuleo G, deFeo G, Scarsella G: Effect of different whole body hyperthermic sessions on the heat shock re-sponse in mice liver and brain. Mol Cell Biochem 204: 41–47, 2000

    Google Scholar 

  26. Cummings CJ, Mancini MA, Antalffy B, DeFranco DB, Orr HT, Zoghbi, HY: Chaperone suppression of aggregation and altered subcellular pro-teasome localization imply protein misfolding in SCA1. Nat Genet 19: 148–154, 1998

    Google Scholar 

  27. Pelham HRB: HSP70 accelerates the recovery of nucleolar morphology after heat shock. EMBO J 3: 3095–3100, 1984

    Google Scholar 

  28. Welch WJ, Suhan JP: Cellular and biochemical events in mammalian cells during and after recovery from physiological stress. J Cell Biol 103: 2035–2053, 1986

    Google Scholar 

  29. Rubin GM, Hogness DS: Effects of heat shock on low molecular weight RNAs in Drosophila melanogaster. Accumulation of a novel form of 5S RNA. Cell 6: 207–213, 1975

    Google Scholar 

  30. Welch WJ: The mammalian stress response: Cell physiology and biochemistry of stress proteins. In: R.J. Morimoto, A. Tissieres, C. Geordopoulos (eds.). Stress Proteins in Biology and Medicine. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 1990, pp. 223–278.

    Google Scholar 

  31. Vogel JL, Parsell DA, Lindquist S: Heat-shock proteins Hsp104 and Hsp70 reactivate mRNA splicing after heat inactivation. Curr Biol 5: 306–317, 1995

    Google Scholar 

  32. Schneiter R, Kadowaki T, Tartakoff AM: mRNA transport in yeast: Time to reinvestigate the functions of the nucleolus. Mol Biol Cell 6: 357–370, 1995

    Google Scholar 

  33. Kadowaki T, Chen S, Hitomi M, Jacobs E, Kumagai C, Liang S, Schneiter R, Singleton D, Wisniewska J, T artakoff AM: Isolation and characterization of Sacharomyces cerevisiae mRNA transport-defective (mtr) mutants. J Cell Biol 126: 649–659, 1994

    Google Scholar 

  34. Kloetzel PM, Bautz EKF: Heat-shock proteins are associated with hnRNA in Drosophila melanogaster tissue culture cells. EMBO J 2: 705–710, 1983

    Google Scholar 

  35. Hietakangas V, Ahlskog JK, Jakobsson AM, Hellesuo M, Sahlberg NM, Holmberg CI, Mikhailov A, P alvimo JJ, Pirkkala L, Sistonen L: Phos-phorylation of serine 303 is a prerequisite for the stress-inducible SUMO modification of heat shock factor 1. Mol Cell Biol 23: 2953–2968, 2003

    Google Scholar 

  36. Teutsch HF, Schuerfeld D, Groezinger E: Three-dimensional recon-struction of parenchymal units in the liver of the rat.Hepatology 29: 494–505, 1999

    Google Scholar 

  37. Schmidt B, Vogelsang M, Haubitz I, Hildebrand R: Lobular distribution pattern of lactate dehydrogenase and 6-phospogluconate dehydrogenase activity in rat liver. Acta Histochimica 102: 37–47, 2000

    Google Scholar 

  38. Hall DM, Oberley TD, Moseley PM, Buettner G, Oberley LW, Weindruch R, Kregel KC: Caloric restriction improves thermotolerance and reduces hyperthermia-induced cellular damage in old rats. FASEB J 14: 76–86, 2000

    Google Scholar 

  39. Manzerra P, Brown JR: The neuronal stress response: Nuclear translo-cation of heat shock proteins as an indicator of hyperthermic stress. Exp Cell Res 229: 35–47, 1996

    Google Scholar 

  40. Mosser DD, Duchaine J, Massie B: The DNA binding activity of the human heat shock transcription factor is regulated in vivo by hsp70. Mol Cell Biol 13: 5427–5438, 1993

    Google Scholar 

  41. Welch WJ, Mizzen LA: Characterization of the thermotolerant cell. II. Effects on the intracellular distribution of heat shock protein 70, intermediate filaments and small ribonucleoprotein complexes. J Cell Biol 106: 1117–1130, 1988

    Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, Institute for Biological Research, Belgrade, Serbia and Montenegro (, Yugoslavia)

    Aleksandra Čvoro & Gordana Matić

  2. Institute of Zoology, Faculty of Biology, University of Belgrade, Serbia and Montenegro (, Yugoslavia)

    Aleksandra Korać

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  1. Aleksandra Čvoro
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  2. Aleksandra Korać
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Čvoro, A., Korać, A. & Matić, G. Intracellular localization of constitutive and inducible heat shock protein 70 in rat liver after in vivo heat stress. Mol Cell Biochem 265, 27–35 (2004). https://doi.org/10.1023/B:MCBI.0000044312.59958.c8

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