Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1990 Apr 1;267(1):125–132. doi: 10.1042/bj2670125

The human heat-shock protein family. Expression of a novel heat-inducible HSP70 (HSP70B') and isolation of its cDNA and genomic DNA.

T K Leung 1, M Y Rajendran 1, C Monfries 1, C Hall 1, L Lim 1
PMCID: PMC1131254  PMID: 2327978

Abstract

The human heat-shock protein multigene family comprises several highly conserved proteins with structural and functional properties in common, but which vary in the extent of their inducibility in response to metabolic stress. We have isolated and characterized a novel human HSP70 cDNA, HSP70B' cDNA, and its corresponding gene sequence. HSP70B' cDNA hybrid-selected an mRNA encoding a more basic 70 kDa heat-shock protein that both the major stress-inducible HSP70 and constitutively expressed HSC70 heat-shock proteins, which in common with other heat-shock 70 kDa proteins bound ATP. The complete HSP70B' gene was sequenced and, like the major inducible HSP70 gene, is devoid of introns. The HSP70B' gene has 77% sequence similarity to the HSP70 gene and 70% similarity to HSC70 cDNA, with greatest sequence divergence towards the 3'-terminus. The HSP70B' gene represents a functional gene, as indicated by Northern-blot analysis with specific oligonucleotides, hybrid-selected translation with a specific 3' cDNA sequence and S1 nuclease protection experiments. In contrast with HSP70 mRNA, which is present at low concentrations in HeLa cells and readily induced by heat or CdCl2 treatment in both fibroblasts and HeLa cells, HSP70B' mRNA was induced only at higher temperature and showed no basal expression. The differences in patterns of induction may be due to the special features of the promoter region of the HSP70B' gene.

Full text

PDF
125

Images in this article

126Fig. 1.
on p.126
130Fig. 4.
on p.130
131Fig. 6.
on p.131

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amin J., Ananthan J., Voellmy R. Key features of heat shock regulatory elements. Mol Cell Biol. 1988 Sep;8(9):3761–3769. doi: 10.1128/mcb.8.9.3761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson N. L., Giometti C. S., Gemmell M. A., Nance S. L., Anderson N. G. A two-dimensional electrophoretic analysis of the heat-shock-induced proteins of human cells. Clin Chem. 1982 Apr;28(4 Pt 2):1084–1092. [PubMed] [Google Scholar]
  3. Birnstiel M. L., Busslinger M., Strub K. Transcription termination and 3' processing: the end is in site! Cell. 1985 Jun;41(2):349–359. doi: 10.1016/s0092-8674(85)80007-6. [DOI] [PubMed] [Google Scholar]
  4. Brown M. E., Amin J., Schiller P., Voellmy R., Scott W. A. Determinants for the DNase I-hypersensitive chromatin structure 5' to a human HSP70 gene. J Mol Biol. 1988 Sep 5;203(1):107–117. doi: 10.1016/0022-2836(88)90095-2. [DOI] [PubMed] [Google Scholar]
  5. Dworniczak B., Mirault M. E. Structure and expression of a human gene coding for a 71 kd heat shock 'cognate' protein. Nucleic Acids Res. 1987 Jul 10;15(13):5181–5197. doi: 10.1093/nar/15.13.5181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Goate A. M., Cooper D. N., Hall C., Leung T. K., Solomon E., Lim L. Localization of a human heat-shock HSP 70 gene sequence to chromosome 6 and detection of two other loci by somatic-cell hybrid and restriction fragment length polymorphism analysis. Hum Genet. 1987 Feb;75(2):123–128. doi: 10.1007/BF00591072. [DOI] [PubMed] [Google Scholar]
  7. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Greene J. M., Larin Z., Taylor I. C., Prentice H., Gwinn K. A., Kingston R. E. Multiple basal elements of a human hsp70 promoter function differently in human and rodent cell lines. Mol Cell Biol. 1987 Oct;7(10):3646–3655. doi: 10.1128/mcb.7.10.3646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hall C., Lowndes C. M., Leung T. K., Cooper D. N., Goate A. M., Lim L. Expression and developmental regulation of two unique mRNAs specific to brain membrane-bound polyribosomes. Biochem J. 1987 Jun 1;244(2):359–366. doi: 10.1042/bj2440359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hall C., Mahadevan L., Whatley S., Biswas G., Lim L. Characterization of translation products of the polyadenylated RNA of free and membrane-bound polyribosomes of rat forebrain. Biochem J. 1984 May 1;219(3):751–761. doi: 10.1042/bj2190751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Harrison G. S., Drabkin H. A., Kao F. T., Hartz J., Hart I. M., Chu E. H., Wu B. J., Morimoto R. I. Chromosomal location of human genes encoding major heat-shock protein HSP70. Somat Cell Mol Genet. 1987 Mar;13(2):119–130. doi: 10.1007/BF01534692. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Kaplan B. B., Bernstein S. L., Gioio A. E. An improved method for the rapid isolation of brain ribonucleic acid. Biochem J. 1979 Oct 1;183(1):181–184. doi: 10.1042/bj1830181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. LeFranc M. P., Forster A., Baer R., Stinson M. A., Rabbitts T. H. Diversity and rearrangement of the human T cell rearranging gamma genes: nine germ-line variable genes belonging to two subgroups. Cell. 1986 Apr 25;45(2):237–246. doi: 10.1016/0092-8674(86)90388-0. [DOI] [PubMed] [Google Scholar]
  16. Lim L., Hall C., Leung T., Whatley S. The relationship of the rat brain 68 kDa microtubule-associated protein with synaptosomal plasma membranes and with the Drosophila 70 kDa heat-shock protein. Biochem J. 1984 Dec 1;224(2):677–680. doi: 10.1042/bj2240677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lindquist S., Craig E. A. The heat-shock proteins. Annu Rev Genet. 1988;22:631–677. doi: 10.1146/annurev.ge.22.120188.003215. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Luckow B., Schütz G. CAT constructions with multiple unique restriction sites for the functional analysis of eukaryotic promoters and regulatory elements. Nucleic Acids Res. 1987 Jul 10;15(13):5490–5490. doi: 10.1093/nar/15.13.5490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Moore M., Schaack J., Baim S. B., Morimoto R. I., Shenk T. Induced heat shock mRNAs escape the nucleocytoplasmic transport block in adenovirus-infected HeLa cells. Mol Cell Biol. 1987 Dec;7(12):4505–4512. doi: 10.1128/mcb.7.12.4505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mues G. I., Munn T. Z., Raese J. D. A human gene family with sequence homology to Drosophila melanogaster Hsp70 heat shock genes. J Biol Chem. 1986 Jan 15;261(2):874–877. [PubMed] [Google Scholar]
  22. Munro S., Pelham H. R. An Hsp70-like protein in the ER: identity with the 78 kd glucose-regulated protein and immunoglobulin heavy chain binding protein. Cell. 1986 Jul 18;46(2):291–300. doi: 10.1016/0092-8674(86)90746-4. [DOI] [PubMed] [Google Scholar]
  23. Napolitano E. W., Pachter J. S., Liem R. K. Intracellular distribution of mammalian stress proteins. Effects of cytoskeletal-specific agents. J Biol Chem. 1987 Feb 5;262(4):1493–1504. [PubMed] [Google Scholar]
  24. 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]
  25. Pelham H. R., Bienz M. A synthetic heat-shock promoter element confers heat-inducibility on the herpes simplex virus thymidine kinase gene. EMBO J. 1982;1(11):1473–1477. doi: 10.1002/j.1460-2075.1982.tb01340.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Richter W. W., Issinger O. G. Differential heat shock response of primary human cell cultures and established cell lines. Biochem Biophys Res Commun. 1986 Nov 26;141(1):46–52. doi: 10.1016/s0006-291x(86)80332-1. [DOI] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schiller P., Amin J., Ananthan J., Brown M. E., Scott W. A., Voellmy R. Cis-acting elements involved in the regulated expression of a human HSP70 gene. J Mol Biol. 1988 Sep 5;203(1):97–105. doi: 10.1016/0022-2836(88)90094-0. [DOI] [PubMed] [Google Scholar]
  29. Sorger P. K., Pelham H. R. Purification and characterization of a heat-shock element binding protein from yeast. EMBO J. 1987 Oct;6(10):3035–3041. doi: 10.1002/j.1460-2075.1987.tb02609.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Wang C., Gomer R. H., Lazarides E. Heat shock proteins are methylated in avian and mammalian cells. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3531–3535. doi: 10.1073/pnas.78.6.3531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Watowich S. S., Morimoto R. I. Complex regulation of heat shock- and glucose-responsive genes in human cells. Mol Cell Biol. 1988 Jan;8(1):393–405. doi: 10.1128/mcb.8.1.393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Whatley S. A., Hall C., Davison A. N., Lim L. Alterations in the relative amounts of specific mRNA species in the developing human brain in Down's syndrome. Biochem J. 1984 May 15;220(1):179–187. doi: 10.1042/bj2200179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Whatley S. A., Leung T., Hall C., Lim L. The brain 68-kilodalton microtubule-associated protein is a cognate form of the 70-kilodalton mammalian heat-shock protein and is present as a specific isoform in synaptosomal membranes. J Neurochem. 1986 Nov;47(5):1576–1583. doi: 10.1111/j.1471-4159.1986.tb00797.x. [DOI] [PubMed] [Google Scholar]
  36. Wu B. J., Kingston R. E., Morimoto R. I. Human HSP70 promoter contains at least two distinct regulatory domains. Proc Natl Acad Sci U S A. 1986 Feb;83(3):629–633. doi: 10.1073/pnas.83.3.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wu B., Hunt C., Morimoto R. Structure and expression of the human gene encoding major heat shock protein HSP70. Mol Cell Biol. 1985 Feb;5(2):330–341. doi: 10.1128/mcb.5.2.330. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES