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  Involvement of transducer of regulated cAMP response element-binding protein activity on corticotropin releasing hormone transcription

Liu, Y., Coello, A. G., Grinevich, V., & Aguilera, G. (2010). Involvement of transducer of regulated cAMP response element-binding protein activity on corticotropin releasing hormone transcription. Endocrinology, 151(3), 1109-1118. doi:10.1210/en.2009-0963.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-59FB-7 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-59FC-5
Genre: Journal Article
Alternative Title : Involvement of transducer of regulated cAMP response element-binding protein activity on corticotropin releasing hormone transcription

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Liu, Yuhong1, Author              
Coello, Anna G., Author
Grinevich, Valery2, Author              
Aguilera, Greti, Author
Affiliations:
1Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497701              
2Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497704              

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 Abstract: We have recently shown that phospho-cAMP response element-binding protein (CREB) is essential but not sufficient for activation of CRH transcription, suggesting the requirement of a coactivator. Here, we test the hypothesis that the CREB coactivator, transducer of regulated CREB activity (TORC), is required for activation of CRH transcription, using the cell line 4B and primary cultures of hypothalamic neurons. Immunohistochemistry and Western blot experiments in 4B cells revealed time-dependent nuclear translocation of TORC1,TORC 2, and TORC3 by forskolin [but not by the phorbol ester, phorbol 12-myristate 13-acetate (PMA)] in a concentration-dependent manner. In reporter gene assays, cotransfection of TORC1 or TORC2 potentiated the stimulatory effect of forskolin on CRH promoter activity but had no effect in cells treated with PMA. Knockout of endogenous TORC using silencing RNA markedly inhibited forskolin-activated CRH promoter activity in 4B cells, as well as the induction of endogenous CRH primary transcript by forskolin in primary neuronal cultures. Coimmunoprecipitation and chromatin immunoprecipitation experiments in 4B cells revealed association of CREB and TORC in the nucleus, and recruitment of TORC2 by the CRH promoter, after 20-min incubation with forskolin. These studies demonstrate a correlation between nuclear translocation of TORC with association to the CRH promoter and activation of CRH transcription. The data suggest that TORC is required for transcriptional activation of the CRH promoter by acting as a CREB coactivator. In addition, cytoplasmic retention of TORC during PMA treatment is likely to explain the failure of phorbolesters to activate CRH transcription in spite of efficiently phosphorylating CREB.

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Language(s): eng - English
 Dates: 2009-08-142009-12-112013-07-012010-03-01
 Publication Status: Published in print
 Pages: 10
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 Rev. Method: Peer
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Title: Endocrinology
Source Genre: Journal
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Publ. Info: Los Angeles, Calif. : Association for the Study of Internal Secretions
Pages: - Volume / Issue: 151 (3) Sequence Number: - Start / End Page: 1109 - 1118 Identifier: ISSN: 0013-7227
CoNE: /journals/resource/991042745737620