Genetic complementation of a glucocorticoid receptor deficiency by expression of cloned receptor cDNA

Roger Miesfeld; Sandro Rusconi; Paul J. Godowski; Bonnie A. Maler; Sam Okret; Ann Charlotte Wikström; Jan Åke Gustafsson; Keith R. Yamamoto

doi:10.1016/0092-8674(86)90659-8

Genetic complementation of a glucocorticoid receptor deficiency by expression of cloned receptor cDNA

Roger Miesfeld
, Sandro Rusconi
, Paul J. Godowski
, Bonnie A. Maler
, Sam Okret
, Ann Charlotte Wikström
, Jan Åke Gustafsson
, Keith R. Yamamoto

Research output: Contribution to journal › Article › peer-review

585 Scopus citations

Abstract

We isolated and sequenced 6.3 kb of cDNA encoding the rat glucocorticoid receptor, a protein that binds and activates a class of hormone-dependent transcriptional enhancers. Receptor-containing cells produce receptor mRNAs of ≅6.5 kb and ≅4.8 kb that differ only in their 3′ nontranslated regions; an open reading frame of 795 amino acids resides within the 5′ portion of the transcripts. The coding region was expressed in vitro, in transient transfections, and in stable transfectants of a receptor-deficient cell line. The protein products are indistinguishable from bona fide receptor with respect to sedimentation and electrophoretic mobility, antibody reactivity, and hormone and DNA binding. Moreover, the cloned receptor protein activates its corresponding enhancers, restoring to the receptor-deficient cells the full capacity for regulated enhancement.

Original language	English (US)
Pages (from-to)	389-399
Number of pages	11
Journal	Cell
Volume	46
Issue number	3
DOIs	https://doi.org/10.1016/0092-8674(86)90659-8
State	Published - Aug 1 1986
Externally published	Yes

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1016/0092-8674(86)90659-8

Cite this

@article{da171cf704a64b068b03d11ca66c601a,

title = "Genetic complementation of a glucocorticoid receptor deficiency by expression of cloned receptor cDNA",

abstract = "We isolated and sequenced 6.3 kb of cDNA encoding the rat glucocorticoid receptor, a protein that binds and activates a class of hormone-dependent transcriptional enhancers. Receptor-containing cells produce receptor mRNAs of ≅6.5 kb and ≅4.8 kb that differ only in their 3′ nontranslated regions; an open reading frame of 795 amino acids resides within the 5′ portion of the transcripts. The coding region was expressed in vitro, in transient transfections, and in stable transfectants of a receptor-deficient cell line. The protein products are indistinguishable from bona fide receptor with respect to sedimentation and electrophoretic mobility, antibody reactivity, and hormone and DNA binding. Moreover, the cloned receptor protein activates its corresponding enhancers, restoring to the receptor-deficient cells the full capacity for regulated enhancement.",

author = "Roger Miesfeld and Sandro Rusconi and Godowski, \{Paul J.\} and Maler, \{Bonnie A.\} and Sam Okret and Wikstr{\"o}m, \{Ann Charlotte\} and Gustafsson, \{Jan {\AA}ke\} and Yamamoto, \{Keith R.\}",

note = "Funding Information: We thank Rae Lyn Burke for pSV7d. Hsiao-Ping Moore for pRSVTG, Bob Harrison for monoclonal antibody BUGR2, and Gordon Ringold for MSN6.10 cells and for communicating results prior to publication. We are indebted to Dan Chin, Steve DiNardo, Rick Myers, and Dennis Sakai for helpful comments on the manuscript, and we acknowledge Kathleen Rafteses for its expert preparation. This work was supported by grants from the National Science Foundation, the National Institutes of Health, and the Swedish Medical Research Council; postdoctoral fellowship support was from the Jane Coffin Childs Memorial Fund for Medical Research (R. M.), the Swiss National Research Foundation (S. R.), and the National Institutes of Health (P J. G.); K. R. Y. is recipient of a Teacher-Scholar award from the Henry and Camille Dreyfus Foundation.",

year = "1986",

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doi = "10.1016/0092-8674(86)90659-8",

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pages = "389--399",

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T1 - Genetic complementation of a glucocorticoid receptor deficiency by expression of cloned receptor cDNA

AU - Miesfeld, Roger

AU - Rusconi, Sandro

AU - Godowski, Paul J.

AU - Maler, Bonnie A.

AU - Okret, Sam

AU - Wikström, Ann Charlotte

AU - Gustafsson, Jan Åke

AU - Yamamoto, Keith R.

N1 - Funding Information: We thank Rae Lyn Burke for pSV7d. Hsiao-Ping Moore for pRSVTG, Bob Harrison for monoclonal antibody BUGR2, and Gordon Ringold for MSN6.10 cells and for communicating results prior to publication. We are indebted to Dan Chin, Steve DiNardo, Rick Myers, and Dennis Sakai for helpful comments on the manuscript, and we acknowledge Kathleen Rafteses for its expert preparation. This work was supported by grants from the National Science Foundation, the National Institutes of Health, and the Swedish Medical Research Council; postdoctoral fellowship support was from the Jane Coffin Childs Memorial Fund for Medical Research (R. M.), the Swiss National Research Foundation (S. R.), and the National Institutes of Health (P J. G.); K. R. Y. is recipient of a Teacher-Scholar award from the Henry and Camille Dreyfus Foundation.

PY - 1986/8/1

Y1 - 1986/8/1

N2 - We isolated and sequenced 6.3 kb of cDNA encoding the rat glucocorticoid receptor, a protein that binds and activates a class of hormone-dependent transcriptional enhancers. Receptor-containing cells produce receptor mRNAs of ≅6.5 kb and ≅4.8 kb that differ only in their 3′ nontranslated regions; an open reading frame of 795 amino acids resides within the 5′ portion of the transcripts. The coding region was expressed in vitro, in transient transfections, and in stable transfectants of a receptor-deficient cell line. The protein products are indistinguishable from bona fide receptor with respect to sedimentation and electrophoretic mobility, antibody reactivity, and hormone and DNA binding. Moreover, the cloned receptor protein activates its corresponding enhancers, restoring to the receptor-deficient cells the full capacity for regulated enhancement.

AB - We isolated and sequenced 6.3 kb of cDNA encoding the rat glucocorticoid receptor, a protein that binds and activates a class of hormone-dependent transcriptional enhancers. Receptor-containing cells produce receptor mRNAs of ≅6.5 kb and ≅4.8 kb that differ only in their 3′ nontranslated regions; an open reading frame of 795 amino acids resides within the 5′ portion of the transcripts. The coding region was expressed in vitro, in transient transfections, and in stable transfectants of a receptor-deficient cell line. The protein products are indistinguishable from bona fide receptor with respect to sedimentation and electrophoretic mobility, antibody reactivity, and hormone and DNA binding. Moreover, the cloned receptor protein activates its corresponding enhancers, restoring to the receptor-deficient cells the full capacity for regulated enhancement.

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Genetic complementation of a glucocorticoid receptor deficiency by expression of cloned receptor cDNA

Abstract

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