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. 1995 Feb;15(2):1071–1078. doi: 10.1128/mcb.15.2.1071

In vivo growth of a murine lymphoma cell line alters regulation of expression of HSP72.

S Davidson 1, P Høj 1, T Gabriele 1, R L Anderson 1
PMCID: PMC232009  PMID: 7823922

Abstract

We have identified a murine B-cell lymphoma cell line, CH1, that has a much-diminished capacity to express increased levels of heat shock proteins in response to heat stress in vitro. In particular, these cells cannot synthesize the inducible 72-kDa heat shock protein (HSP72) which is normally expressed at high levels in stressed cells. We show here that CH1 fails to transcribe HSP72 mRNA after heat shock, even though the heat shock transcription factor, HSF, is activated correctly. After heat shock, HSF from CH1 is found in the nucleus and is phosphorylated, trimerized, and capable of binding the heat shock element. We propose that additional signals which CH1 cells are unable to transduce are normally required to activate hsp72 transcription in vitro. Surprisingly, we have found that when the CH1 cells are heated in situ in a mouse, they show normal expression of HSP72 mRNA and protein. Therefore, CH1 cells have a functional hsp72 gene which can be transcribed and translated when the cells are in an appropriate environment. A diffusible factor present in ascites fluid is capable of restoring normal HSP72 induction in CH1 cells. We conclude that as-yet-undefined factors are required for regulation of the hsp72 gene or, alternatively, that heat shock in vivo causes activation of hsp70 through a novel pathway which the defect in CH1 has exposed and which is distinct from that operating in vitro. This unique system offers an opportunity to study a physiologically relevant pathway of heat shock induction and to biochemically define effectors involved in the mammalian stress response.

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Selected References

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  1. Abravaya K., Myers M. P., Murphy S. P., Morimoto R. I. The human heat shock protein hsp70 interacts with HSF, the transcription factor that regulates heat shock gene expression. Genes Dev. 1992 Jul;6(7):1153–1164. doi: 10.1101/gad.6.7.1153. [DOI] [PubMed] [Google Scholar]
  2. Agoff S. N., Hou J., Linzer D. I., Wu B. Regulation of the human hsp70 promoter by p53. Science. 1993 Jan 1;259(5091):84–87. doi: 10.1126/science.8418500. [DOI] [PubMed] [Google Scholar]
  3. Anderson R. L., Van Kersen I., Kraft P. E., Hahn G. M. Biochemical analysis of heat-resistant mouse tumor cell strains: a new member of the HSP70 family. Mol Cell Biol. 1989 Aug;9(8):3509–3516. doi: 10.1128/mcb.9.8.3509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Anderson R. L., Wang C. Y., van Kersen I., Lee K. J., Welch W. J., Lavagnini P., Hahn G. M. An immunoassay for heat shock protein 73/72: use of the assay to correlate HSP73/72 levels in mammalian cells with heat response. Int J Hyperthermia. 1993 Jul-Aug;9(4):539–552. doi: 10.3109/02656739309005051. [DOI] [PubMed] [Google Scholar]
  5. Aujame L. The major heat-shock protein hsp 68 is not induced by stress in mouse erythroleukemia cell lines. Biochem Cell Biol. 1988 Jul;66(7):691–701. doi: 10.1139/o88-079. [DOI] [PubMed] [Google Scholar]
  6. Baler R., Dahl G., Voellmy R. Activation of human heat shock genes is accompanied by oligomerization, modification, and rapid translocation of heat shock transcription factor HSF1. Mol Cell Biol. 1993 Apr;13(4):2486–2496. doi: 10.1128/mcb.13.4.2486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Blake M. J., Udelsman R., Feulner G. J., Norton D. D., Holbrook N. J. Stress-induced heat shock protein 70 expression in adrenal cortex: an adrenocorticotropic hormone-sensitive, age-dependent response. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9873–9877. doi: 10.1073/pnas.88.21.9873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bruce J. L., Price B. D., Coleman C. N., Calderwood S. K. Oxidative injury rapidly activates the heat shock transcription factor but fails to increase levels of heat shock proteins. Cancer Res. 1993 Jan 1;53(1):12–15. [PubMed] [Google Scholar]
  9. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  10. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fisher B., Kraft P., Hahn G. M., Anderson R. L. Thermotolerance in the absence of induced heat shock proteins in a murine lymphoma. Cancer Res. 1992 May 15;52(10):2854–2861. [PubMed] [Google Scholar]
  12. Foos G., Natour S., Klempnauer K. H. TATA-box dependent trans-activation of the human HSP70 promoter by Myb proteins. Oncogene. 1993 Jul;8(7):1775–1782. [PubMed] [Google Scholar]
  13. Gething M. J., Sambrook J. Protein folding in the cell. Nature. 1992 Jan 2;355(6355):33–45. doi: 10.1038/355033a0. [DOI] [PubMed] [Google Scholar]
  14. Giebel L. B., Dworniczak B. P., Bautz E. K. Developmental regulation of a constitutively expressed mouse mRNA encoding a 72-kDa heat shock-like protein. Dev Biol. 1988 Jan;125(1):200–207. doi: 10.1016/0012-1606(88)90073-5. [DOI] [PubMed] [Google Scholar]
  15. Hahn G. M., Ning S. C., Elizaga M., Kapp D. S., Anderson R. L. A comparison of thermal responses of human and rodent cells. Int J Radiat Biol. 1989 Nov;56(5):817–825. doi: 10.1080/09553008914552101. [DOI] [PubMed] [Google Scholar]
  16. Haughton G., Arnold L. W., Bishop G. A., Mercolino T. J. The CH series of murine B cell lymphomas: neoplastic analogues of Ly-1+ normal B cells. Immunol Rev. 1986 Oct;93:35–51. doi: 10.1111/j.1600-065x.1986.tb01501.x. [DOI] [PubMed] [Google Scholar]
  17. Hensold J. O., Hunt C. R., Calderwood S. K., Housman D. E., Kingston R. E. DNA binding of heat shock factor to the heat shock element is insufficient for transcriptional activation in murine erythroleukemia cells. Mol Cell Biol. 1990 Apr;10(4):1600–1608. doi: 10.1128/mcb.10.4.1600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hickey E., Brandon S. E., Sadis S., Smale G., Weber L. A. Molecular cloning of sequences encoding the human heat-shock proteins and their expression during hyperthermia. Gene. 1986;43(1-2):147–154. doi: 10.1016/0378-1119(86)90018-1. [DOI] [PubMed] [Google Scholar]
  19. Hunt C., Calderwood S. Characterization and sequence of a mouse hsp70 gene and its expression in mouse cell lines. Gene. 1990 Mar 15;87(2):199–204. doi: 10.1016/0378-1119(90)90302-8. [DOI] [PubMed] [Google Scholar]
  20. Jurivich D. A., Sistonen L., Kroes R. A., Morimoto R. I. Effect of sodium salicylate on the human heat shock response. Science. 1992 Mar 6;255(5049):1243–1245. doi: 10.1126/science.1546322. [DOI] [PubMed] [Google Scholar]
  21. Kobayashi N., McEntee K. Identification of cis and trans components of a novel heat shock stress regulatory pathway in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Jan;13(1):248–256. doi: 10.1128/mcb.13.1.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Langer T., Lu C., Echols H., Flanagan J., Hayer M. K., Hartl F. U. Successive action of DnaK, DnaJ and GroEL along the pathway of chaperone-mediated protein folding. Nature. 1992 Apr 23;356(6371):683–689. doi: 10.1038/356683a0. [DOI] [PubMed] [Google Scholar]
  23. Lanier L. L., Lynes M., Haughton G., Wettstein P. J. Novel type of murine B-cell lymphoma. Nature. 1978 Feb 9;271(5645):554–555. doi: 10.1038/271554a0. [DOI] [PubMed] [Google Scholar]
  24. Lindquist S. The heat-shock response. Annu Rev Biochem. 1986;55:1151–1191. doi: 10.1146/annurev.bi.55.070186.005443. [DOI] [PubMed] [Google Scholar]
  25. Milarski K. L., Welch W. J., Morimoto R. I. Cell cycle-dependent association of HSP70 with specific cellular proteins. J Cell Biol. 1989 Feb;108(2):413–423. doi: 10.1083/jcb.108.2.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Morange M., Diu A., Bensaude O., Babinet C. Altered expression of heat shock proteins in embryonal carcinoma and mouse early embryonic cells. Mol Cell Biol. 1984 Apr;4(4):730–735. doi: 10.1128/mcb.4.4.730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Morimoto R. I., Sarge K. D., Abravaya K. Transcriptional regulation of heat shock genes. A paradigm for inducible genomic responses. J Biol Chem. 1992 Nov 5;267(31):21987–21990. [PubMed] [Google Scholar]
  28. Mosser D. D., Duchaine J., Massie B. The DNA-binding activity of the human heat shock transcription factor is regulated in vivo by hsp70. Mol Cell Biol. 1993 Sep;13(9):5427–5438. doi: 10.1128/mcb.13.9.5427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nakai A., Morimoto R. I. Characterization of a novel chicken heat shock transcription factor, heat shock factor 3, suggests a new regulatory pathway. Mol Cell Biol. 1993 Apr;13(4):1983–1997. doi: 10.1128/mcb.13.4.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Perry M. D., Moran L. A. Isolation of a mouse heat-shock gene (hsp68) by recombinational screening. Gene. 1987;51(2-3):227–236. doi: 10.1016/0378-1119(87)90311-8. [DOI] [PubMed] [Google Scholar]
  31. Rabindran S. K., Haroun R. I., Clos J., Wisniewski J., Wu C. Regulation of heat shock factor trimer formation: role of a conserved leucine zipper. Science. 1993 Jan 8;259(5092):230–234. doi: 10.1126/science.8421783. [DOI] [PubMed] [Google Scholar]
  32. Sarge K. D., Murphy S. P., Morimoto R. I. Activation of heat shock gene transcription by heat shock factor 1 involves oligomerization, acquisition of DNA-binding activity, and nuclear localization and can occur in the absence of stress. Mol Cell Biol. 1993 Mar;13(3):1392–1407. doi: 10.1128/mcb.13.3.1392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sistonen L., Sarge K. D., Phillips B., Abravaya K., Morimoto R. I. Activation of heat shock factor 2 during hemin-induced differentiation of human erythroleukemia cells. Mol Cell Biol. 1992 Sep;12(9):4104–4111. doi: 10.1128/mcb.12.9.4104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Spector N. L., Freedman A. S., Freeman G., Segil J., Whitman J. F., Welch W. J., Nadler L. M. Activation primes human B lymphocytes to respond to heat shock. J Exp Med. 1989 Nov 1;170(5):1763–1768. doi: 10.1084/jem.170.5.1763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Twentyman P. R., Brown J. M., Gray J. W., Franko A. J., Scoles M. A., Kallman R. F. A new mouse tumor model system (RIF-1) for comparison of end-point studies. J Natl Cancer Inst. 1980 Mar;64(3):595–604. [PubMed] [Google Scholar]
  36. Welch W. J. Mammalian stress response: cell physiology, structure/function of stress proteins, and implications for medicine and disease. Physiol Rev. 1992 Oct;72(4):1063–1081. doi: 10.1152/physrev.1992.72.4.1063. [DOI] [PubMed] [Google Scholar]
  37. Welch W. J., Mizzen L. A. Characterization of the thermotolerant cell. II. Effects on the intracellular distribution of heat-shock protein 70, intermediate filaments, and small nuclear ribonucleoprotein complexes. J Cell Biol. 1988 Apr;106(4):1117–1130. doi: 10.1083/jcb.106.4.1117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wettstein P. J., Haughton G. Production, testing, and utility of double congenic strains of mice.I. B10 H-2aa=H-48P-wis and Bio-H-4BP-Wis. Transplantation. 1974 May;17(5):513–517. doi: 10.1097/00007890-197405000-00012. [DOI] [PubMed] [Google Scholar]
  39. Williams G. T., McClanahan T. K., Morimoto R. I. E1a transactivation of the human HSP70 promoter is mediated through the basal transcriptional complex. Mol Cell Biol. 1989 Jun;9(6):2574–2587. doi: 10.1128/mcb.9.6.2574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wittig S., Hensse S., Keitel C., Elsner C., Wittig B. Heat shock gene expression is regulated during teratocarcinoma cell differentiation and early embryonic development. Dev Biol. 1983 Apr;96(2):507–514. doi: 10.1016/0012-1606(83)90187-2. [DOI] [PubMed] [Google Scholar]

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