Characterization of progesterone receptor binding to the 90- and 70- kDa heat shock proteins

D. B. Schowalter, W. P. Sullivan, Nita Jane Maihle, A. D.W. Dobson, O. M. Conneely, B. W. O'Malley, D. O. Toft

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

In this study a model system for expression of the chicken progesterone receptor in cultured cells was developed using a quail fibroblast cell line, QT6. The chicken progesterone receptor form A expressed in QT6 cells was evaluated and determined to have a number of similarities to receptor isolated from chicken oviduct. These include hormone binding, sedimentation profile, phosphorylation pattern, heat shock protein (hsp) 70 and hsp90 associations and the ability to stimulate a reporter gene construct. Therefore, the receptor expressed in this system functioned adequately for further evaluation of the particular region (or regions) involved in hsp70 and hsp90 binding. Several receptor deletion mutants were tested for hsp70/ hsp90 binding; only the d369-659 mutant, which has the entire steroid-binding domain deleted, was unable to bind hsp90 and hsp70. Three separate regions of the steroid-binding domain were found to partially restore hsp90 and hsp70 binding to the d369-659 mutant protein. However, hsp binding was not abolished when these or other regions of the steroid binding domain were deleted individually. These findings indicate that hsp90 and hsp70 both bind to the steroid-binding domain of the receptor through interactions at multiple locations or through some structural quality that is distributed throughout this region of the protein.

Original languageEnglish (US)
Pages (from-to)21165-21173
Number of pages9
JournalJournal of Biological Chemistry
Volume266
Issue number31
StatePublished - Jan 1 1991
Externally publishedYes

Fingerprint

HSP70 Heat-Shock Proteins
Progesterone Receptors
Steroids
Chickens
Cells
Phosphorylation
Quail
Oviducts
Mutant Proteins
Fibroblasts
Heat-Shock Proteins
Reporter Genes
Sedimentation
Protein Binding
Cultured Cells
Genes
Hormones
Cell Line
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Schowalter, D. B., Sullivan, W. P., Maihle, N. J., Dobson, A. D. W., Conneely, O. M., O'Malley, B. W., & Toft, D. O. (1991). Characterization of progesterone receptor binding to the 90- and 70- kDa heat shock proteins. Journal of Biological Chemistry, 266(31), 21165-21173.

Characterization of progesterone receptor binding to the 90- and 70- kDa heat shock proteins. / Schowalter, D. B.; Sullivan, W. P.; Maihle, Nita Jane; Dobson, A. D.W.; Conneely, O. M.; O'Malley, B. W.; Toft, D. O.

In: Journal of Biological Chemistry, Vol. 266, No. 31, 01.01.1991, p. 21165-21173.

Research output: Contribution to journalArticle

Schowalter, DB, Sullivan, WP, Maihle, NJ, Dobson, ADW, Conneely, OM, O'Malley, BW & Toft, DO 1991, 'Characterization of progesterone receptor binding to the 90- and 70- kDa heat shock proteins', Journal of Biological Chemistry, vol. 266, no. 31, pp. 21165-21173.
Schowalter DB, Sullivan WP, Maihle NJ, Dobson ADW, Conneely OM, O'Malley BW et al. Characterization of progesterone receptor binding to the 90- and 70- kDa heat shock proteins. Journal of Biological Chemistry. 1991 Jan 1;266(31):21165-21173.
Schowalter, D. B. ; Sullivan, W. P. ; Maihle, Nita Jane ; Dobson, A. D.W. ; Conneely, O. M. ; O'Malley, B. W. ; Toft, D. O. / Characterization of progesterone receptor binding to the 90- and 70- kDa heat shock proteins. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 31. pp. 21165-21173.
@article{4f6a2d7c1bbd4c2c95a55b1eec0e5259,
title = "Characterization of progesterone receptor binding to the 90- and 70- kDa heat shock proteins",
abstract = "In this study a model system for expression of the chicken progesterone receptor in cultured cells was developed using a quail fibroblast cell line, QT6. The chicken progesterone receptor form A expressed in QT6 cells was evaluated and determined to have a number of similarities to receptor isolated from chicken oviduct. These include hormone binding, sedimentation profile, phosphorylation pattern, heat shock protein (hsp) 70 and hsp90 associations and the ability to stimulate a reporter gene construct. Therefore, the receptor expressed in this system functioned adequately for further evaluation of the particular region (or regions) involved in hsp70 and hsp90 binding. Several receptor deletion mutants were tested for hsp70/ hsp90 binding; only the d369-659 mutant, which has the entire steroid-binding domain deleted, was unable to bind hsp90 and hsp70. Three separate regions of the steroid-binding domain were found to partially restore hsp90 and hsp70 binding to the d369-659 mutant protein. However, hsp binding was not abolished when these or other regions of the steroid binding domain were deleted individually. These findings indicate that hsp90 and hsp70 both bind to the steroid-binding domain of the receptor through interactions at multiple locations or through some structural quality that is distributed throughout this region of the protein.",
author = "Schowalter, {D. B.} and Sullivan, {W. P.} and Maihle, {Nita Jane} and Dobson, {A. D.W.} and Conneely, {O. M.} and O'Malley, {B. W.} and Toft, {D. O.}",
year = "1991",
month = "1",
day = "1",
language = "English (US)",
volume = "266",
pages = "21165--21173",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "31",

}

TY - JOUR

T1 - Characterization of progesterone receptor binding to the 90- and 70- kDa heat shock proteins

AU - Schowalter, D. B.

AU - Sullivan, W. P.

AU - Maihle, Nita Jane

AU - Dobson, A. D.W.

AU - Conneely, O. M.

AU - O'Malley, B. W.

AU - Toft, D. O.

PY - 1991/1/1

Y1 - 1991/1/1

N2 - In this study a model system for expression of the chicken progesterone receptor in cultured cells was developed using a quail fibroblast cell line, QT6. The chicken progesterone receptor form A expressed in QT6 cells was evaluated and determined to have a number of similarities to receptor isolated from chicken oviduct. These include hormone binding, sedimentation profile, phosphorylation pattern, heat shock protein (hsp) 70 and hsp90 associations and the ability to stimulate a reporter gene construct. Therefore, the receptor expressed in this system functioned adequately for further evaluation of the particular region (or regions) involved in hsp70 and hsp90 binding. Several receptor deletion mutants were tested for hsp70/ hsp90 binding; only the d369-659 mutant, which has the entire steroid-binding domain deleted, was unable to bind hsp90 and hsp70. Three separate regions of the steroid-binding domain were found to partially restore hsp90 and hsp70 binding to the d369-659 mutant protein. However, hsp binding was not abolished when these or other regions of the steroid binding domain were deleted individually. These findings indicate that hsp90 and hsp70 both bind to the steroid-binding domain of the receptor through interactions at multiple locations or through some structural quality that is distributed throughout this region of the protein.

AB - In this study a model system for expression of the chicken progesterone receptor in cultured cells was developed using a quail fibroblast cell line, QT6. The chicken progesterone receptor form A expressed in QT6 cells was evaluated and determined to have a number of similarities to receptor isolated from chicken oviduct. These include hormone binding, sedimentation profile, phosphorylation pattern, heat shock protein (hsp) 70 and hsp90 associations and the ability to stimulate a reporter gene construct. Therefore, the receptor expressed in this system functioned adequately for further evaluation of the particular region (or regions) involved in hsp70 and hsp90 binding. Several receptor deletion mutants were tested for hsp70/ hsp90 binding; only the d369-659 mutant, which has the entire steroid-binding domain deleted, was unable to bind hsp90 and hsp70. Three separate regions of the steroid-binding domain were found to partially restore hsp90 and hsp70 binding to the d369-659 mutant protein. However, hsp binding was not abolished when these or other regions of the steroid binding domain were deleted individually. These findings indicate that hsp90 and hsp70 both bind to the steroid-binding domain of the receptor through interactions at multiple locations or through some structural quality that is distributed throughout this region of the protein.

UR - http://www.scopus.com/inward/record.url?scp=0025872036&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025872036&partnerID=8YFLogxK

M3 - Article

C2 - 1939158

AN - SCOPUS:0025872036

VL - 266

SP - 21165

EP - 21173

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 31

ER -