Home LiteratureArticle Details
PMID: 2527334 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Sequence and regulation of a gene encoding a human 89-kilodalton heat shock protein.

Molecular and cellular biology ·Vol. 9 ·No. 6 ·1989-06-00 ·Pages 2615-26

Hickey E, Brandon SE, Smale G, Lloyd D, Weber LA

Abstract

Vertebrate cells synthesize two forms of the 82- to 90-kilodalton heat shock protein that are encoded by distinct gene families. In HeLa cells, both proteins (hsp89 alpha and hsp89 beta) are abundant under normal growth conditions and are synthesized at increased rates in response to heat stress. Only the larger form, hsp89 alpha, is induced by the adenovirus E1A gene product (M. C. Simon, K. Kitchener, H. T. Kao, E. Hickey, L. Weber, R. Voellmy, N. Heintz, and J. R. Nevins, Mol. Cell. Biol. 7:2884-2890, 1987). We have isolated a human hsp89 alpha gene that shows complete sequence identity with heat- and E1A-inducible cDNA used as a hybridization probe. The 5'-flanking region contained overlapping and inverted consensus heat shock control elements that can confer heat-inducible expression on a beta-globin reporter gene. The gene contained 10 intervening sequences. The first intron was located adjacent to the translation start codon, an arrangement also found in the Drosophila hsp82 gene. The spliced mRNA sequence contained a single open reading frame encoding an 84,564-dalton polypeptide showing high homology with the hsp82 to hsp90 proteins of other organisms. The deduced hsp89 alpha protein sequence differed from the human hsp89 beta sequence reported elsewhere (N. F. Rebbe, J. Ware, R. M. Bertina, P. Modrich, and D. W. Stafford (Gene 53:235-245, 1987) in at least 99 out of the 732 amino acids. Transcription of the hsp89 alpha gene was induced by serum during normal cell growth, but expression did not appear to be restricted to a particular stage of the cell cycle. hsp89 alpha mRNA was considerably more stable than the mRNA encoding hsp70, which can account for the higher constitutive rate of hsp89 synthesis in unstressed cells.

MeSH Terms
Adenovirus Early Proteins Amino Acid Sequence Base Sequence Blood Blotting, Southern Cell Line Cloning, Molecular DNA/genetics Gene Expression Regulation Genes Heat-Shock Proteins/genetics Humans Molecular Sequence Data Molecular Weight Multigene Family Oncogene Proteins, Viral/physiology Promoter Regions, Genetic RNA, Messenger/genetics Restriction Mapping Transfection
Chemicals
Adenovirus Early Proteins Heat-Shock Proteins Oncogene Proteins, Viral RNA, Messenger DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Hickey E
Biology Department, University of South Florida, Tampa 33620.
Brandon S E
Smale G
Lloyd D
Weber L A
References (83)
83 references, click to expand
  1. Biochemical characterization of the mammalian stress proteins and identification of two stress proteins as glucose- and Ca2+-ionophore-regulated proteins.
    J Biol Chem. 1983 Jun 10;258(11):7102-11 PMID: 6406494
  2. Regulation of human histone gene expression: kinetics of accumulation and changes in the rate of synthesis and in the half-lives of individual histone mRNAs during the HeLa cell cycle.
    Mol Cell Biol. 1983 Apr;3(4):539-50 PMID: 6406835
  3. Coordinate regulation of multiple histone mRNAs during the cell cycle in HeLa cells.
    Nucleic Acids Res. 1983 Apr 25;11(8):2391-410 PMID: 6304651
  4. New M13 vectors for cloning.
    Methods Enzymol. 1983;101:20-78 PMID: 6310323
  5. Metabolite regulation of heat shock protein levels.
    Proc Natl Acad Sci U S A. 1983 Sep;80(17):5325-9 PMID: 6577432
  6. Mouse and Drosophila genes encoding the major heat shock protein (hsp70) are highly conserved.
    Mol Cell Biol. 1983 Aug;3(8):1540-3 PMID: 6413838
  7. Changes in liver-specific compared to common gene transcription during primary culture of mouse hepatocytes.
    Mol Cell Biol. 1983 Sep;3(9):1552-61 PMID: 6633533
  8. Regulation of histone mRNA production and stability in serum-stimulated mouse 3T6 fibroblasts.
    Mol Cell Biol. 1983 Nov;3(11):1920-9 PMID: 6656760
  9. DNA sequence analysis on the IBM-PC.
    Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):601-4 PMID: 6546432
  10. A DNA sequence analysis package for the IBM personal computer.
    Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):605-14 PMID: 6546433
  11. Complete sequence of the heat shock-inducible HSP90 gene of Saccharomyces cerevisiae.
    J Biol Chem. 1984 May 10;259(9):5745-51 PMID: 6325446
  12. Molecular cloning of a steroid-regulated 108K heat shock protein gene from hen oviduct.
    Nucleic Acids Res. 1986 Dec 22;14(24):10053-69 PMID: 3027654
  13. The genome of Trypanosoma cruzi contains a constitutively expressed, tandemly arranged multicopy gene homologous to a major heat shock protein.
    Mol Cell Biol. 1987 Mar;7(3):1271-5 PMID: 3550435
  14. Detection of three protein binding sites in the serum-regulated promoter of the human gene encoding the 70-kDa heat shock protein.
    Proc Natl Acad Sci U S A. 1987 Apr;84(8):2203-7 PMID: 3031671
  15. Mouse 89 kD heat shock protein. Two polypeptides with distinct developmental regulation.
    Exp Cell Res. 1987 May;170(1):186-94 PMID: 3569431
  16. Relationship of the 90-kDa murine heat shock protein to the untransformed and transformed states of the L cell glucocorticoid receptor.
    J Biol Chem. 1987 May 25;262(15):6986-91 PMID: 3294824
  17. Expression of intron-containing C. elegans heat shock genes in mouse cells demonstrates divergence of 3' splice site recognition sequences between nematodes and vertebrates, and an inhibitory effect of heat shock on the mammalian splicing apparatus.
    Nucleic Acids Res. 1987 May 11;15(9):3723-41 PMID: 3588308
  18. Cloning and expression of a gene encoding hsc73, the major hsp70-like protein in unstressed rat cells.
    EMBO J. 1987 Apr;6(4):993-8 PMID: 3595567
  19. Structure and expression of a human gene coding for a 71 kd heat shock 'cognate' protein.
    Nucleic Acids Res. 1987 Jul 10;15(13):5181-97 PMID: 3037489
  20. Evidence that the 90-kilodalton heat shock protein is associated with tubulin-containing complexes in L cell cytosol and in intact PtK cells.
    Mol Endocrinol. 1988 Aug;2(8):756-60 PMID: 3062385
  21. Eukaryotic Mr 83,000 heat shock protein has a homologue in Escherichia coli.
    Proc Natl Acad Sci U S A. 1987 Aug;84(15):5177-81 PMID: 3299380
  22. Subcellular localization of the 84,000 dalton heat-shock protein in mouse neuroblastoma cells: evidence for a cytoplasmic and nuclear location.
    Eur J Cell Biol. 1987 Jun;43(3):469-78 PMID: 3305023
  23. Electrophoresis in agarose and acrylamide gels.
    Methods Enzymol. 1987;152:61-87 PMID: 2443811
  24. Mapping of gene transcripts by nuclease protection assays and cDNA primer extension.
    Methods Enzymol. 1987;152:611-32 PMID: 2821358
  25. RNA splice junctions of different classes of eukaryotes: sequence statistics and functional implications in gene expression.
    Nucleic Acids Res. 1987 Sep 11;15(17):7155-74 PMID: 3658675
  26. Transformation of glucocorticoid and progesterone receptors to the DNA-binding state.
    J Cell Biochem. 1987 Sep;35(1):51-68 PMID: 3312247
  27. Selective induction of human heat shock gene transcription by the adenovirus E1A gene products, including the 12S E1A product.
    Mol Cell Biol. 1987 Aug;7(8):2884-90 PMID: 2959854
  28. Cloning and nucleotide sequence of the murine hsp84 cDNA and chromosome assignment of related sequences.
    Gene. 1987;56(1):29-40 PMID: 2445630
  29. The 90-kilodalton peptide of the heme-regulated eIF-2 alpha kinase has sequence similarity with the 90-kilodalton heat shock protein.
    Biochemistry. 1987 Oct 20;26(21):6583-7 PMID: 3427028
  30. Steroid hormone antagonists at the receptor level: a role for the heat-shock protein MW 90,000 (hsp 90).
    J Cell Biochem. 1987 Oct;35(2):161-74 PMID: 3323207
  31. Posttranscriptional regulation of hsp70 expression in human cells: effects of heat shock, inhibition of protein synthesis, and adenovirus infection on translation and mRNA stability.
    Mol Cell Biol. 1987 Dec;7(12):4357-68 PMID: 3437893
  32. Germline transformation used to define key features of heat-shock response elements.
    Science. 1988 Mar 4;239(4844):1139-42 PMID: 3125608
  33. Mitogen activation induces the enhanced synthesis of two heat-shock proteins in human lymphocytes.
    J Cell Biol. 1988 Mar;106(3):883-91 PMID: 3346328
  34. Sequence analysis of the nonsteroid binding component of the calf uterine estrogen receptor.
    Biochem Biophys Res Commun. 1988 Mar 30;151(3):1156-63 PMID: 3355547
  35. Effect of the 90 kDa heat shock protein, HSP90, on glucocorticoid receptor binding to DNA-cellulose.
    Biochem Biophys Res Commun. 1988 Mar 30;151(3):1226-32 PMID: 3355552
  36. Estrogenic regulation of uterine 90-kilodalton heat shock protein.
    Endocrinology. 1988 Aug;123(2):956-61 PMID: 3293991
  37. Effect of heat shock on RNA metabolism in HeLa cells.
    J Cell Physiol. 1988 Jun;135(3):377-86 PMID: 2456286
  38. Heat shock but not other stress inducers leads to the disruption of a sub-set of snRNPs and inhibition of in vitro splicing in HeLa cells.
    EMBO J. 1988 Nov;7(11):3509-18 PMID: 2974799
  39. Nucleotide sequence of a cDNA for a member of the human 90-kDa heat-shock protein family.
    Gene. 1987;53(2-3):235-45 PMID: 3301534
  40. Two human 90-kDa heat shock proteins are phosphorylated in vivo at conserved serines that are phosphorylated in vitro by casein kinase II.
    J Biol Chem. 1989 Feb 15;264(5):2431-7 PMID: 2492519
  41. Altered expression of heat shock proteins in embryonal carcinoma and mouse early embryonic cells.
    Mol Cell Biol. 1984 Apr;4(4):730-5 PMID: 6546970
  42. Common non-hormone binding component in non-transformed chick oviduct receptors of four steroid hormones.
    Nature. 1984 Apr 26-May 2;308(5962):850-3 PMID: 6201744
  43. Xenopus hsp 70 genes are constitutively expressed in injected oocytes.
    EMBO J. 1984 Nov;3(11):2477-83 PMID: 6510409
  44. Quantitation and intracellular localization of the 85K heat shock protein by using monoclonal and polyclonal antibodies.
    Mol Cell Biol. 1984 Dec;4(12):2802-10 PMID: 6396506
  45. Double-stranded DNA induces the phosphorylation of several proteins including the 90 000 mol. wt. heat-shock protein in animal cell extracts.
    EMBO J. 1985 Jan;4(1):139-45 PMID: 4018025
  46. Locus encoding a family of small heat shock genes in Caenorhabditis elegans: two genes duplicated to form a 3.8-kilobase inverted repeat.
    Mol Cell Biol. 1985 Jun;5(6):1268-78 PMID: 4033652
  47. Transcription of the human hsp70 gene is induced by serum stimulation.
    Proc Natl Acad Sci U S A. 1985 Sep;82(18):6070-4 PMID: 3862119
  48. 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):6455-9 PMID: 3931075
  49. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes.
    Cell. 1986 Jan 31;44(2):283-92 PMID: 3943125
  50. Molecular cloning and analysis of DNA complementary to three mouse Mr = 68,000 heat shock protein mRNAs.
    J Biol Chem. 1986 Feb 15;261(5):2102-12 PMID: 2868009
  51. Human HSP70 promoter contains at least two distinct regulatory domains.
    Proc Natl Acad Sci U S A. 1986 Feb;83(3):629-33 PMID: 3456160
  52. RNA splicing is interrupted by heat shock and is rescued by heat shock protein synthesis.
    Cell. 1986 Apr 25;45(2):185-93 PMID: 2421918
  53. A mouse tumor-specific transplantation antigen is a heat shock-related protein.
    Proc Natl Acad Sci U S A. 1986 May;83(10):3121-5 PMID: 3458168
  54. Sequence and organization of genes encoding the human 27 kDa heat shock protein.
    Nucleic Acids Res. 1986 May 27;14(10):4127-45 PMID: 3714473
  55. Interspecific nucleotide sequence comparisons used to identify regulatory and structural features of the Drosophila hsp82 gene.
    J Mol Biol. 1986 Apr 20;188(4):499-515 PMID: 2426456
  56. The heat-shock response.
    Annu Rev Biochem. 1986;55:1151-91 PMID: 2427013
  57. Translational activation of maternal mRNA encoding the heat-shock protein hsp90 during sea urchin embryogenesis.
    Dev Biol. 1986 Sep;117(1):286-93 PMID: 3743897
  58. Association of the heat shock protein hsp90 with steroid hormone receptors and tyrosine kinase oncogene products.
    Biochem Biophys Res Commun. 1986 Aug 14;138(3):1298-307 PMID: 3530253
  59. Molecular cloning of sequences encoding the human heat-shock proteins and their expression during hyperthermia.
    Gene. 1986;43(1-2):147-54 PMID: 3019832
  60. Cellular localization of Drosophila 83-kilodalton heat shock protein in normal, heat-shocked, and recovering cultured cells with a specific antibody.
    Biochem Cell Biol. 1986 Aug;64(8):816-25 PMID: 2429681
  61. The dynamic state of heat shock proteins in chicken embryo fibroblasts.
    J Cell Biol. 1986 Oct;103(4):1495-507 PMID: 3533955
  62. Two mammalian heat shock proteins, HSP90 and HSP100, are actin-binding proteins.
    Proc Natl Acad Sci U S A. 1986 Nov;83(21):8054-8 PMID: 3534880
  63. Calmodulin-regulated binding of the 90-kDa heat shock protein to actin filaments.
    J Biol Chem. 1986 Dec 5;261(34):16033-6 PMID: 3782106
  64. The chicken ubiquitin gene contains a heat shock promoter and expresses an unstable mRNA in heat-shocked cells.
    Mol Cell Biol. 1986 Dec;6(12):4602-10 PMID: 3025663
  65. Conserved sequences and transcription of the hsp70 gene family in Trypanosoma brucei.
    Mol Cell Biol. 1986 Dec;6(12):4657-66 PMID: 3796613
  66. A new technique for the assay of infectivity of human adenovirus 5 DNA.
    Virology. 1973 Apr;52(2):456-67 PMID: 4705382
  67. Detection of specific sequences among DNA fragments separated by gel electrophoresis.
    J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
  68. A general method for isolation of high molecular weight DNA from eukaryotes.
    Nucleic Acids Res. 1976 Sep;3(9):2303-8 PMID: 987581
  69. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I.
    J Mol Biol. 1977 Jun 15;113(1):237-51 PMID: 881736
  70. Isolation of histone messenger RNA and its translation in vitro.
    Methods Cell Biol. 1978;19:215-36 PMID: 567734
  71. The isolation and characterization of linked delta- and beta-globin genes from a cloned library of human DNA.
    Cell. 1978 Dec;15(4):1157-74 PMID: 728996
  72. Different responses to drugs and serum of cells transformed by various means.
    Cancer Res. 1979 Jul;39(7 Pt 1):2718-26 PMID: 445475
  73. Purification and characterization of a major component from the cytoplasmic matrix of cultured murine L cells.
    Biochim Biophys Acta. 1979 May 23;578(1):1-12 PMID: 454659
  74. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping.
    Methods Enzymol. 1980;65(1):718-49 PMID: 6154876
  75. The nucleotide sequence of the human beta-globin gene.
    Cell. 1980 Oct;21(3):647-51 PMID: 6254664
  76. Sequence of three copies of the gene for the major Drosophila heat shock induced protein and their flanking regions.
    Cell. 1980 Oct;21(3):669-79 PMID: 6777045
  77. A cellular protein that associates with the transforming protein of Rous sarcoma virus is also a heat-shock protein.
    Proc Natl Acad Sci U S A. 1981 Feb;78(2):1067-71 PMID: 6262754
  78. The specific interaction of the Rous sarcoma virus transforming protein, pp60src, with two cellular proteins.
    Cell. 1981 Aug;25(2):363-72 PMID: 6269742
  79. Modulation of heat-shock polypeptide synthesis in HeLa cells during hyperthermia and recovery.
    Biochemistry. 1982 Mar 30;21(7):1513-21 PMID: 7082633
  80. Antibodies to two major chicken heat shock proteins cross-react with similar proteins in widely divergent species.
    Mol Cell Biol. 1982 Mar;2(3):267-74 PMID: 7110134
  81. A regulatory upstream promoter element in the Drosophila hsp 70 heat-shock gene.
    Cell. 1982 Sep;30(2):517-28 PMID: 6814763
  82. Purification of the major mammalian heat shock proteins.
    J Biol Chem. 1982 Dec 25;257(24):14949-59 PMID: 7174676
  83. Interaction between the Rous sarcoma virus transforming protein and two cellular phosphoproteins: analysis of the turnover and distribution of this complex.
    Mol Cell Biol. 1983 Jan;3(1):9-19 PMID: 6298609
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.9.6.2615-2626.1989
Published
1989-06-00
Pages
2615-26
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC362334
Subset
IM
Grants
NCI NIH HHS · CA36207 · United States
NIGMS NIH HHS · GM32381 · United States
Databases
GENBANK
M27024
External Links
PubMed source DOI: 10.1128/mcb.9.6.2615-2626.1989 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2