A chemical biology screen identifies glucocorticoids that regulate c-maf expression by increasing its proteasomal degradation through up-regulation of ubiquitin

Xinliang Mao, Alexander Keith Stewart, Rose Hurren, Alessandro Datti, Xuegong Zhu, Yuanxiao Zhu, Changxin Shi, Kyle Lee, Rodger Tiedemann, Yanina Eberhard, Suzanne Trudel, Shengben Liang, Seth J. Corey, Lisa C. Gillis, Dwayne L. Barber, Jeffery L. Wrana, Shereen Ezzat, Aaron D. Schimmer

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Abstract

The oncogene c-maf is frequently overexpressed in multiple myeloma cell lines and patient samples and contributes to increased cellular proliferation in part by inducing cyclin D2 expression. To identify regulators of c-maf, we developed a chemical screen in NIH3T3 cells stably overexpressing c-maf and the cyclin D2 promoter driving luciferase. From a screen of 2400 off-patent drugs and chemicals, we identified glucocorticoids as c-maf-dependent inhibitors of cyclin D2 transactivation. In multiple myeloma cell lines, glucocorticoids reduced levels of c-maf protein without influencing corresponding mRNA levels. Subsequent studies demonstrated that glucocorticoids increased ubiquitination-dependent degradation of c-maf and up-regulated ubiquitin C mRNA. Moreover, ectopic expression of ubiquitin C recapitulated the effects of glucocorticoids, demonstrating regulation of c-maf protein through the abundance of the ubiquitin substrate. Thus, using a chemical biology approach, we identified a novel mechanism of action of glucocorticoids and a novel mechanism by which levels of c-maf protein are regulated by the abundance of the ubiquitin substrate.

Original languageEnglish (US)
Pages (from-to)4047-4054
Number of pages8
JournalBlood
Volume110
Issue number12
DOIs
StatePublished - Dec 1 2007

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Proto-Oncogene Proteins c-maf
Ubiquitin
Glucocorticoids
Cyclin D2
Up-Regulation
Ubiquitin C
Degradation
Multiple Myeloma
Cells
Cell Line
Messenger RNA
Ubiquitination
Substrates
Luciferases
Oncogenes
Transcriptional Activation
Cell Proliferation
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Hematology

Cite this

A chemical biology screen identifies glucocorticoids that regulate c-maf expression by increasing its proteasomal degradation through up-regulation of ubiquitin. / Mao, Xinliang; Stewart, Alexander Keith; Hurren, Rose; Datti, Alessandro; Zhu, Xuegong; Zhu, Yuanxiao; Shi, Changxin; Lee, Kyle; Tiedemann, Rodger; Eberhard, Yanina; Trudel, Suzanne; Liang, Shengben; Corey, Seth J.; Gillis, Lisa C.; Barber, Dwayne L.; Wrana, Jeffery L.; Ezzat, Shereen; Schimmer, Aaron D.

In: Blood, Vol. 110, No. 12, 01.12.2007, p. 4047-4054.

Research output: Contribution to journalArticle

Mao, X, Stewart, AK, Hurren, R, Datti, A, Zhu, X, Zhu, Y, Shi, C, Lee, K, Tiedemann, R, Eberhard, Y, Trudel, S, Liang, S, Corey, SJ, Gillis, LC, Barber, DL, Wrana, JL, Ezzat, S & Schimmer, AD 2007, 'A chemical biology screen identifies glucocorticoids that regulate c-maf expression by increasing its proteasomal degradation through up-regulation of ubiquitin', Blood, vol. 110, no. 12, pp. 4047-4054. https://doi.org/10.1182/blood-2007-05-088666
Mao, Xinliang ; Stewart, Alexander Keith ; Hurren, Rose ; Datti, Alessandro ; Zhu, Xuegong ; Zhu, Yuanxiao ; Shi, Changxin ; Lee, Kyle ; Tiedemann, Rodger ; Eberhard, Yanina ; Trudel, Suzanne ; Liang, Shengben ; Corey, Seth J. ; Gillis, Lisa C. ; Barber, Dwayne L. ; Wrana, Jeffery L. ; Ezzat, Shereen ; Schimmer, Aaron D. / A chemical biology screen identifies glucocorticoids that regulate c-maf expression by increasing its proteasomal degradation through up-regulation of ubiquitin. In: Blood. 2007 ; Vol. 110, No. 12. pp. 4047-4054.
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abstract = "The oncogene c-maf is frequently overexpressed in multiple myeloma cell lines and patient samples and contributes to increased cellular proliferation in part by inducing cyclin D2 expression. To identify regulators of c-maf, we developed a chemical screen in NIH3T3 cells stably overexpressing c-maf and the cyclin D2 promoter driving luciferase. From a screen of 2400 off-patent drugs and chemicals, we identified glucocorticoids as c-maf-dependent inhibitors of cyclin D2 transactivation. In multiple myeloma cell lines, glucocorticoids reduced levels of c-maf protein without influencing corresponding mRNA levels. Subsequent studies demonstrated that glucocorticoids increased ubiquitination-dependent degradation of c-maf and up-regulated ubiquitin C mRNA. Moreover, ectopic expression of ubiquitin C recapitulated the effects of glucocorticoids, demonstrating regulation of c-maf protein through the abundance of the ubiquitin substrate. Thus, using a chemical biology approach, we identified a novel mechanism of action of glucocorticoids and a novel mechanism by which levels of c-maf protein are regulated by the abundance of the ubiquitin substrate.",
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AU - Zhu, Yuanxiao

AU - Shi, Changxin

AU - Lee, Kyle

AU - Tiedemann, Rodger

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AU - Liang, Shengben

AU - Corey, Seth J.

AU - Gillis, Lisa C.

AU - Barber, Dwayne L.

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AU - Ezzat, Shereen

AU - Schimmer, Aaron D.

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AB - The oncogene c-maf is frequently overexpressed in multiple myeloma cell lines and patient samples and contributes to increased cellular proliferation in part by inducing cyclin D2 expression. To identify regulators of c-maf, we developed a chemical screen in NIH3T3 cells stably overexpressing c-maf and the cyclin D2 promoter driving luciferase. From a screen of 2400 off-patent drugs and chemicals, we identified glucocorticoids as c-maf-dependent inhibitors of cyclin D2 transactivation. In multiple myeloma cell lines, glucocorticoids reduced levels of c-maf protein without influencing corresponding mRNA levels. Subsequent studies demonstrated that glucocorticoids increased ubiquitination-dependent degradation of c-maf and up-regulated ubiquitin C mRNA. Moreover, ectopic expression of ubiquitin C recapitulated the effects of glucocorticoids, demonstrating regulation of c-maf protein through the abundance of the ubiquitin substrate. Thus, using a chemical biology approach, we identified a novel mechanism of action of glucocorticoids and a novel mechanism by which levels of c-maf protein are regulated by the abundance of the ubiquitin substrate.

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