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.
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Tissieres A, Mitchell HK, Tracy U: Protein synthesis in salivary glands ofDrosophila melanogaster. Relation to chromosome puffs. J Mol Biol 84: 389–398, 1974
Peluso RW, Lamb RA, Choppin, PW: Polypeptide synthesis in simian virus 5-infected cells. J Virol 23: 177–187, 1977
Sorger PK, Pelham HRB: Yeast heat shock factor is an essential DNA-binding protein that exhibits temperature-dependent phosphorylation. Cell 54: 855–864, 1988
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
Subjeck JR, Sciandra JJ, Johnson RJ: Heat shock proteins and thermotolerance: A comparison of induction kinetics. Br J Radiol 55: 579–584, 1982
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
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
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
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
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
Horwitz J: α-Crystallin can function as a molecular chaperone. Proc Natl Acad Sci USA 89: 10449–10453, 1992
Jakob U, Gaestel M, Engel K, Buchner J: Small heat shock proteins are molecular chaperones. J Biol Chem 268: 1517–1520, 1993
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
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
Fröhli E, Aoyama A, Klemenz R: Cloning of the mouse hsp25 gene and an extremely conserved hsp25 pseudogene. Gene 128: 273–277, 1993
Gaestel M, Gotthardt R, Milller T: Structure and organisation of a murine gene encoding small heat-shock protein Hsp25. Gene 128: 279–283, 1993
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
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
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
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
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
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
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
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
Wu C: An exonuclease protection assay reveals heat-shock element and TATA-box binding proteins in crude nuclear extracts. Nature 317: 84–87, 1985
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
Kingston RE, Schuetz TJ, Larin Z: Heat inducible human factor that binds to a human hsp70 promoter. Mol Cell Biol 7: 1530–1534, 1987
Sorger PK, Pelham HRB: Purification and characterization of a heat shock element binding protein from yeast. EMBO J 6: 3035–3041, 1987
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
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
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
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
Laszlo A: Evidence for two states of thermotolerance in mammalian cells. Int J Hyperthermia 4: 513–526, 1988
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
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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
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DOI: https://doi.org/10.1007/BF00714333