Home LiteratureArticle Details
PMID: 9584173 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Genetic and biochemical analysis of p23 and ansamycin antibiotics in the function of Hsp90-dependent signaling proteins.

Molecular and cellular biology ·Vol. 18 ·No. 6 ·1998-06-00 ·Pages 3330-9

Bohen SP

Abstract

The ubiquitous molecular chaperone Hsp90 acts in concert with a cohort of associated proteins to facilitate the functional maturation of a number of cellular signaling proteins, such as steroid hormone receptors and oncogene tyrosine kinases. The Hsp90-associated protein p23 is required for the assembly of functional steroid aporeceptor complexes in cell lysates, and Hsp90-binding ansamycin antibiotics disrupt the activity of Hsp90-dependent signaling proteins in cultured mammalian cells and prevent the association of p23 with Hsp90-receptor heterocomplexes; these observations have led to the hypotheses that p23 is required for the maturation of Hsp90 target proteins and that ansamycin antibiotics abrogate the activity of such proteins by disrupting the interaction of p23 with Hsp90. In this study, I demonstrate that ansamycin antibiotics disrupt the function of Hsp90 target proteins expressed in yeast cells; prevent the assembly of Sba1, a yeast p23-like protein, into steroid receptor-Hsp90 complexes; and result in the assembly of receptor-Hsp90 complexes that are defective for ligand binding. To assess the role of p23 in Hsp90 target protein function, I show that the activity of Hsp90 target proteins is unaffected by deletion of SBA1. Interestingly, steroid receptor activity in cells lacking Sba1 displays increased sensitivity to ansamycin antibiotics, and this phenotype is rescued by the expression of human p23 in yeast cells. These findings indicate that Hsp90-dependent signaling proteins can achieve a functional conformation in vivo in the absence of p23. Furthermore, while the presence of p23 decreases the sensitivity of Hsp90-dependent processes to ansamycin treatment, ansamycin antibiotics disrupt signaling through some mechanism other than altering the Hsp90-p23 interaction.

MeSH Terms
Amino Acid Sequence Anti-Bacterial Agents/metabolism Antibiotics, Antineoplastic/pharmacology Benzoquinones DNA-Binding Proteins/genetics,metabolism HSP90 Heat-Shock Proteins/metabolism Humans Lactams, Macrocyclic Molecular Sequence Data Oncogene Protein pp60(v-src)/metabolism Phenotype Protein-Tyrosine Kinases/antagonists & inhibitors,metabolism Quinones/pharmacology Receptors, Steroid/antagonists & inhibitors,metabolism Saccharomyces cerevisiae/drug effects,genetics Sequence Alignment Signal Transduction
Chemicals
Anti-Bacterial Agents Antibiotics, Antineoplastic Benzoquinones DNA-Binding Proteins HSP90 Heat-Shock Proteins Lactams, Macrocyclic Quinones Receptors, Steroid protein p23 macbecin I macbecin II Protein-Tyrosine Kinases Oncogene Protein pp60(v-src) geldanamycin
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Bohen S P
Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892-4255, USA. [email protected]
References (67)
67 references, click to expand
  1. Crystal structure of an Hsp90-geldanamycin complex: targeting of a protein chaperone by an antitumor agent.
    Cell. 1997 Apr 18;89(2):239-50 PMID: 9108479
  2. Folding of the glucocorticoid receptor by the reconstituted Hsp90-based chaperone machinery. The initial hsp90.p60.hsp70-dependent step is sufficient for creating the steroid binding conformation.
    J Biol Chem. 1997 May 16;272(20):13047-54 PMID: 9148915
  3. The function of steroid hormone receptors is inhibited by the hsp90-specific compound geldanamycin.
    J Biol Chem. 1997 Jul 25;272(30):18694-701 PMID: 9228040
  4. The specific DNA binding activity of the dioxin receptor is modulated by the 90 kd heat shock protein.
    EMBO J. 1990 Jan;9(1):69-76 PMID: 2153080
  5. Reduced levels of hsp90 compromise steroid receptor action in vivo.
    Nature. 1990 Nov 8;348(6297):166-8 PMID: 2234079
  6. The viral erbA oncogene protein, a constitutive repressor in animal cells, is a hormone-regulated activator in yeast.
    Cell. 1990 Dec 21;63(6):1277-86 PMID: 1979758
  7. Getting started with yeast.
    Methods Enzymol. 1991;194:3-21 PMID: 2005794
  8. Retinoic acid receptor belongs to a subclass of nuclear receptors that do not form "docking" complexes with hsp90.
    Biochemistry. 1991 Jun 4;30(22):5605-8 PMID: 1645195
  9. Use and selectivity of herbimycin A as inhibitor of protein-tyrosine kinases.
    Methods Enzymol. 1991;201:370-9 PMID: 1658554
  10. Protein components of the nonactivated glucocorticoid receptor.
    J Biol Chem. 1991 Dec 25;266(36):24601-5 PMID: 1761557
  11. Assembly of progesterone receptor with heat shock proteins and receptor activation are ATP mediated events.
    J Biol Chem. 1992 Jan 15;267(2):1350-6 PMID: 1730655
  12. Multifunctional yeast high-copy-number shuttle vectors.
    Gene. 1992 Jan 2;110(1):119-22 PMID: 1544568
  13. Improved method for high efficiency transformation of intact yeast cells.
    Nucleic Acids Res. 1992 Mar 25;20(6):1425 PMID: 1561104
  14. A model of glucocorticoid receptor unfolding and stabilization by a heat shock protein complex.
    J Steroid Biochem Mol Biol. 1992 Mar;41(3-8):223-9 PMID: 1373296
  15. The mechanism of RU486 antagonism is dependent on the conformation of the carboxy-terminal tail of the human progesterone receptor.
    Cell. 1992 May 15;69(4):703-13 PMID: 1586949
  16. A heat shock protein complex isolated from rabbit reticulocyte lysate can reconstitute a functional glucocorticoid receptor-Hsp90 complex.
    Biochemistry. 1992 Aug 18;31(32):7325-9 PMID: 1510923
  17. Heterotetrameric structure of the human progesterone receptor.
    Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8001-5 PMID: 1518826
  18. The translation machinery and 70 kd heat shock protein cooperate in protein synthesis.
    Cell. 1992 Oct 2;71(1):97-105 PMID: 1394434
  19. Genetic dissection of the signaling domain of a mammalian steroid receptor in yeast.
    Mol Biol Cell. 1992 Nov;3(11):1245-57 PMID: 1457829
  20. Steroid receptors and their associated proteins.
    Mol Endocrinol. 1993 Jan;7(1):4-11 PMID: 8446107
  21. Reconstitution of retinoid X receptor function and combinatorial regulation of other nuclear hormone receptors in the yeast Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):6929-33 PMID: 8394003
  22. Heat-shock protein hsp90 governs the activity of pp60v-src kinase.
    Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7074-8 PMID: 7688470
  23. Suppression of c-Src activity by C-terminal Src kinase involves the c-Src SH2 and SH3 domains: analysis with Saccharomyces cerevisiae.
    Mol Cell Biol. 1993 Sep;13(9):5290-300 PMID: 7689149
  24. Isolation of Hsp90 mutants by screening for decreased steroid receptor function.
    Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11424-8 PMID: 8248264
  25. Proof that hsp70 is required for assembly of the glucocorticoid receptor into a heterocomplex with hsp90.
    J Biol Chem. 1994 Feb 18;269(7):5043-9 PMID: 8106480
  26. Identification and structural characterization of the ATP/ADP-binding site in the Hsp90 molecular chaperone.
    Cell. 1997 Jul 11;90(1):65-75 PMID: 9230303
  27. Folding of the glucocorticoid receptor by the heat shock protein (hsp) 90-based chaperone machinery. The role of p23 is to stabilize receptor.hsp90 heterocomplexes formed by hsp90.p60.hsp70.
    J Biol Chem. 1997 Aug 22;272(34):21213-20 PMID: 9261129
  28. The amino-terminal domain of heat shock protein 90 (hsp90) that binds geldanamycin is an ATP/ADP switch domain that regulates hsp90 conformation.
    J Biol Chem. 1997 Sep 19;272(38):23843-50 PMID: 9295332
  29. Dynamics of heat shock protein 90-progesterone receptor binding and the disactivation loop model for steroid receptor complexes.
    Mol Endocrinol. 1993 Nov;7(11):1418-29 PMID: 7906860
  30. Complete DNA sequence of yeast chromosome XI.
    Nature. 1994 Jun 2;369(6479):371-8 PMID: 8196765
  31. Mutations in Hsp83 and cdc37 impair signaling by the sevenless receptor tyrosine kinase in Drosophila.
    Cell. 1994 Jul 1;77(7):1027-36 PMID: 8020093
  32. Inhibition of heat shock protein HSP90-pp60v-src heteroprotein complex formation by benzoquinone ansamycins: essential role for stress proteins in oncogenic transformation.
    Proc Natl Acad Sci U S A. 1994 Aug 30;91(18):8324-8 PMID: 8078881
  33. Conservation of Hsp90 macromolecular complexes in Saccharomyces cerevisiae.
    J Biol Chem. 1994 Oct 7;269(40):24983-8 PMID: 7929182
  34. A novel chaperone complex for steroid receptors involving heat shock proteins, immunophilins, and p23.
    J Biol Chem. 1994 Oct 7;269(40):24989-93 PMID: 7929183
  35. A role for Hsp90 in cell cycle control: Wee1 tyrosine kinase activity requires interaction with Hsp90.
    EMBO J. 1994 Dec 15;13(24):6099-106 PMID: 7813446
  36. LEM1, an ATP-binding-cassette transporter, selectively modulates the biological potency of steroid hormones.
    Proc Natl Acad Sci U S A. 1995 May 9;92(10):4701-5 PMID: 7753868
  37. Growth inhibition of virus transformed cells in vitro and antitumor activity in vivo of geldanamycin and its derivatives.
    J Antibiot (Tokyo). 1979 Aug;32(8):849-51 PMID: 500504
  38. Macbecins I and II, new antitumor antibiotics. I. Producing organism, fermentation and antimicrobial activities.
    J Antibiot (Tokyo). 1980 Feb;33(2):199-204 PMID: 7380729
  39. Antitumor and cytocidal activities of a newly isolated benzenoid ansamycin, macbecin I.
    Gan. 1982 Dec;73(6):938-44 PMID: 6186564
  40. 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
  41. Evidence that the 90-kDa phosphoprotein associated with the untransformed L-cell glucocorticoid receptor is a murine heat shock protein.
    J Biol Chem. 1985 Oct 15;260(23):12398-401 PMID: 3900074
  42. Steroid receptor regulated transcription of specific genes and gene networks.
    Annu Rev Genet. 1985;19:209-52 PMID: 3909942
  43. Interaction of the Rous sarcoma virus protein pp60src with the cellular proteins pp50 and pp90.
    Curr Top Microbiol Immunol. 1986;123:1-22 PMID: 2419040
  44. The common 90-kd protein component of non-transformed '8S' steroid receptors is a heat-shock protein.
    EMBO J. 1985 Dec 1;4(12):3131-5 PMID: 2419124
  45. Glucocorticoid receptor mutants that are constitutive activators of transcriptional enhancement.
    Nature. 1987 Jan 22-28;325(6102):365-8 PMID: 3808033
  46. The steroid and thyroid hormone receptor superfamily.
    Science. 1988 May 13;240(4854):889-95 PMID: 3283939
  47. Mammalian glucocorticoid receptor derivatives enhance transcription in yeast.
    Science. 1988 Aug 19;241(4868):965-7 PMID: 3043665
  48. Evidence that the 90-kDa heat shock protein is necessary for the steroid binding conformation of the L cell glucocorticoid receptor.
    J Biol Chem. 1989 Mar 25;264(9):4992-7 PMID: 2647745
  49. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  50. hsp82 is an essential protein that is required in higher concentrations for growth of cells at higher temperatures.
    Mol Cell Biol. 1989 Sep;9(9):3919-30 PMID: 2674684
  51. Characterization of a novel 23-kilodalton protein of unactive progesterone receptor complexes.
    Mol Cell Biol. 1994 Mar;14(3):1956-63 PMID: 8114727
  52. Hold 'em and fold 'em: chaperones and signal transduction.
    Science. 1995 Jun 2;268(5215):1303-4 PMID: 7761850
  53. Role of the protein chaperone YDJ1 in establishing Hsp90-mediated signal transduction pathways.
    Science. 1995 Jun 2;268(5215):1362-5 PMID: 7761857
  54. Preclinical pharmacologic evaluation of geldanamycin as an antitumor agent.
    Cancer Chemother Pharmacol. 1995;36(4):305-15 PMID: 7628050
  55. The 23-kDa acidic protein in reticulocyte lysate is the weakly bound component of the hsp foldosome that is required for assembly of the glucocorticoid receptor into a functional heterocomplex with hsp90.
    J Biol Chem. 1995 Aug 11;270(32):18841-7 PMID: 7642537
  56. Disruption of the Raf-1-Hsp90 molecular complex results in destabilization of Raf-1 and loss of Raf-1-Ras association.
    J Biol Chem. 1995 Oct 13;270(41):24585-8 PMID: 7592678
  57. Progesterone receptor structure and function altered by geldanamycin, an hsp90-binding agent.
    Mol Cell Biol. 1995 Dec;15(12):6804-12 PMID: 8524246
  58. Hsp90 mutants disrupt glucocorticoid receptor ligand binding and destabilize aporeceptor complexes.
    J Biol Chem. 1995 Dec 8;270(49):29433-8 PMID: 7493981
  59. A role for Hsp90 in retinoid receptor signal transduction.
    Mol Biol Cell. 1995 Dec;6(12):1833-42 PMID: 8590809
  60. Binding of p23 and hsp90 during assembly with the progesterone receptor.
    Mol Endocrinol. 1995 Jun;9(6):670-8 PMID: 8592513
  61. An FK506-sensitive transporter selectively decreases intracellular levels and potency of steroid hormones.
    J Biol Chem. 1996 Jul 19;271(29):17152-6 PMID: 8663352
  62. Mutant conformation of p53 translated in vitro or in vivo requires functional HSP90.
    Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8379-83 PMID: 8710879
  63. Stable and specific binding of heat shock protein 90 by geldanamycin disrupts glucocorticoid receptor function in intact cells.
    Mol Endocrinol. 1996 Jun;10(6):705-12 PMID: 8776730
  64. A cyclophilin function in Hsp90-dependent signal transduction.
    Science. 1996 Dec 6;274(5293):1713-5 PMID: 8939862
  65. In vivo analysis of the Hsp90 cochaperone Sti1 (p60).
    Mol Cell Biol. 1997 Jan;17(1):318-25 PMID: 8972212
  66. Functional specificity among Hsp70 molecular chaperones.
    Science. 1997 Jan 17;275(5298):387-9 PMID: 8994035
  67. Nucleotides and two functional states of hsp90.
    J Biol Chem. 1997 Mar 21;272(12):8007-12 PMID: 9065472
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.18.6.3330
Email
[email protected]
Published
1998-06-00
Pages
3330-9
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC108914
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.18.6.3330 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