Skip to main content
Log in

Thermal response in murine L929 cells lacking αB-crystallin expression and αB-crystallin expressing L929 transfectants

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

We investigated the role of αB-crystallin expression in the development of thermotolerance in murine L929 cells. An initial heat-shock of 10 min at 45°C induced thermotolerance in these cells to a heat challenge at 45°C administered 24 h later. The thermotolerance ratio at 10−1 isosurvival was 1.7. Expression of αB-crystallin gene was not detected during the 24 h incubation at 37°C following heat shock by either northern or western blots. In contrast, inducible HSP70 synthesis was observed during this time period. Thus, this cell line provided an unique system in which to examine the effects of transfected αB-crystallin on thermoresistance and thermotolerance. Cells stably transfected with αB-crystallin under the control of an inducible promoter did not show a significant increase in the ability to develop thermotolerance. However, a stably transfected L929 clone expressing high levels of constitutive αB-crystallin exhibited an approximately 50% increase in thermal resistance over parental and control cells. Though expression of αB-crystallin is not requisite for the development of thermotolerance in L929 cells, overexpression of transfected αB-crystallin can contribute to increased thermoresistance.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Tissieres A, Mitchell HK, Tracy U: Protein synthesis in salivary glands ofDrosophila melanogaster. Relation to chromosome puffs. J Mol Biol 84: 389–398, 1974

    Google Scholar 

  2. Peluso RW, Lamb RA, Choppin, PW: Polypeptide synthesis in simian virus 5-infected cells. J Virol 23: 177–187, 1977

    Google Scholar 

  3. Sorger PK, Pelham HRB: Yeast heat shock factor is an essential DNA-binding protein that exhibits temperature-dependent phosphorylation. Cell 54: 855–864, 1988

    Google Scholar 

  4. Li GC: Correlation between synthesis of heat shock proteins and development of thermotolerance in Chinese hamster fibroblasts. Proc Natl Acad Sci USA 79: 3218–3222, 1982

    Google Scholar 

  5. Subjeck JR, Sciandra JJ, Johnson RJ: Heat shock proteins and thermotolerance: A comparison of induction kinetics. Br J Radiol 55: 579–584, 1982

    Google Scholar 

  6. Kim D, Lee YJ, Corry PM: Employment of a turbidometric assay system to study the biochemical role of HSP70 in heat-induced protein aggregation. J Therm Biol 18: 165–175, 1993

    Google Scholar 

  7. Hahn GM, Li GC: Thermotolerance, thermoresistance, and thermosensitization. In: R.I. Morimoto, A. Tissieres, C. Georgopoulis (eds). Stress Proteins in Biology and Medicine. Cold Spring Harbor Laboratory Press/Cold Spring Harbor, NY, 1990, pp. 79–100

    Google Scholar 

  8. Landry J, Chretien P, Lambert H, Hickey E, Weber LA: Heat shock resistance conferred by expression of the human HSP27 gene in rodent cells. J Cell Biol 109: 7–15, 1989

    Google Scholar 

  9. Klemenz R, Frohli E, Aoyama A, Hoffmann S, Simpson RJ, Moritz RL, Schafer RL: αB crystallin accumulation is a specific response to Ha-ras and v-mos oncogene expression in mouse NIH 3T3 fibroblasts. Mol Cell Biol 11: 803–812, 1991a

    Google Scholar 

  10. Klemenz R, Frohli E, Steiger RH, Schafer R, Aoyama A: αB-crystallin is a small heat shock protein. Proc Natl Acad Sci USA 88: 3652–3656, 1991b

    Google Scholar 

  11. Horwitz J: α-Crystallin can function as a molecular chaperone. Proc Natl Acad Sci USA 89: 10449–10453, 1992

    Google Scholar 

  12. Jakob U, Gaestel M, Engel K, Buchner J: Small heat shock proteins are molecular chaperones. J Biol Chem 268: 1517–1520, 1993

    Google Scholar 

  13. Aoyama A, Frohli E, Schafer R, Klemenz R: αB-crystallin expression in mouse NIH 3T3 fibroblasts: glucocorticoid responsiveness and involvement in thermal protection. Mol Cell Biol 13: 1824–1835, 1993

    Google Scholar 

  14. Mehlen P, Preville X, Chareyron P, Briolay J, Klemenz R, Arrigo AP: Constitutive expression of human hsp27,Drosophila hsp27, human αB-crystallin confers resistance to TNF- and oxidative stress-induced cytotoxicity in stably transfected murine L929 fibroblasts. J Immunol 154: 363–374, 1995

    Google Scholar 

  15. Fröhli E, Aoyama A, Klemenz R: Cloning of the mouse hsp25 gene and an extremely conserved hsp25 pseudogene. Gene 128: 273–277, 1993

    Google Scholar 

  16. Gaestel M, Gotthardt R, Milller T: Structure and organisation of a murine gene encoding small heat-shock protein Hsp25. Gene 128: 279–283, 1993

    Google Scholar 

  17. Hickey E, Brandon SE, Potter R, Stein G, Stein J, Weber LA: Sequence and organization of genes encoding the human 27 kDa heat shock protein. Nucl Acids Res 14: 4127–4145, 1986

    Google Scholar 

  18. Lee YJ, Hou Z, Curetty L, Borrelli MJ: Development of acute thermotolerance in L929 cells: lack of HSP28 synthesis and phosphorylation. J Cell Physiol 152: 118–125, 1992a

    Google Scholar 

  19. Lee YJ, Hou Z, Curetty L, Borrelli MJ, Corry PM: Absence of HSP28 sythesis and phosphorylation during the development of chronic thermotolerance in murine L929 cells. Cancer Res 52: 5780–5787, 1992b

    Google Scholar 

  20. Ingolia TD, Craig EA: Four smallDrosophila heat shock proteins are related to each other and to mammalian α-crystallin. Proc Natl Acad Sci USA 79: 2360–2364, 1982

    Google Scholar 

  21. Tushinski R, Sussman P, Yu L, Bancroft F: Pregrowth hormone messenger RNA: Glucocorticoid induction and identification in rat pituitary cells. Proc Nat Acad Sci USA 74: 2357–2361, 1977

    Google Scholar 

  22. Lehrach H, Diamond L, Wozney J, Boedtker H: RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry 16: 4743–4751, 1977

    Google Scholar 

  23. Gunning P, Leavitt J, Muscat G, Ng S-Y, Kedes L: A human β-actin expression vector system directs high-level accumulation of antisense transcripts. Proc Natl Acad Sci USA 84: 4831–4835, 1987

    Google Scholar 

  24. Parker CS, Topol JA:Drosophila RNA polymerase II transcription factor binds to the regulatory site of an lisp 70 gene. Cell 37: 273–283, 1984

    Google Scholar 

  25. Wu C: An exonuclease protection assay reveals heat-shock element and TATA-box binding proteins in crude nuclear extracts. Nature 317: 84–87, 1985

    Google Scholar 

  26. Wu C, Wilson S, Walker B, David I, Paisley T, Zimarino V, Ueda H: Purification and properties ofDrosophila heat shock activator protein. Science 238: 1247–1253, 1987

    Google Scholar 

  27. Kingston RE, Schuetz TJ, Larin Z: Heat inducible human factor that binds to a human hsp70 promoter. Mol Cell Biol 7: 1530–1534, 1987

    Google Scholar 

  28. Sorger PK, Pelham HRB: Purification and characterization of a heat shock element binding protein from yeast. EMBO J 6: 3035–3041, 1987

    Google Scholar 

  29. Wiederrecht G, Shuey DJ, Kibbe WA, Parker CS: The Saccharomyces and Drosophila heat shock transcription factors are identical in size and DNA binding properties. Cell 48: 507–515, 1987

    Google Scholar 

  30. Li GC: Induction of thermotolerance and enhanced heat shock protein synthesis in Chinese hamster fibroblasts by sodium arsenite and by ethanol. J Cell Physiol 115: 116–122, 1983

    Google Scholar 

  31. Lee YJ, Hou Z-Z, Curetty L, Erdos G, Stromberg JS, Carper SW, Cho JM, Corry PM: Regulation of HSP70 and HSP28 gene expression: absence of compensatory interactions. Mol Cell Biochem 137: 155–167, 1994

    Google Scholar 

  32. Li GC, Li L, Liu Y-K, Mak JY, Chen L, Lee WMF: Thermal response of rat fibroblasts stably transfected with the human 70-kDa heat shock protein-encoding gene. Proc Natl Acad Sci USA 88: 1681–1685, 1991

    Google Scholar 

  33. Laszlo A: Evidence for two states of thermotolerance in mammalian cells. Int J Hyperthermia 4: 513–526, 1988

    Google Scholar 

  34. Lee YJ, Dewey WC: Thermotolerance induced by heat, sodium arsenite, or puromycin: Its inhibition and differences between 43°C and 45°C. J Cell Physiol 135: 397–406, 1988

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Blackburn, R., Galoforo, S., Berns, C.M. et al. Thermal response in murine L929 cells lacking αB-crystallin expression and αB-crystallin expressing L929 transfectants. Mol Cell Biochem 155, 51–60 (1996). https://doi.org/10.1007/BF00714333

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00714333

Key words

Navigation