CRHR1-dependent effects on protein expression and posttranslational modification in AtT-20 cells

Helena C. Kronsbein; Archana M. Jastorff; Giuseppina Maccarrone; Günter Stalla; Wolfgang Wurst; Florian Holsboer; Christoph W. Turck; Jan M. Deussing

doi:10.1016/j.mce.2008.05.017

CRHR1-dependent effects on protein expression and posttranslational modification in AtT-20 cells

Helena C. Kronsbein
, Archana M. Jastorff
, Giuseppina Maccarrone
, Günter Stalla
, Wolfgang Wurst
, Florian Holsboer
, Christoph W. Turck
, Jan M. Deussing

Max Planck Institute of Psychiatry
Helmholtz Zentrum München German Research Center for Environmental Health

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

6 Scopus citations

Abstract

Corticotropin-releasing hormone (CRH) plays a major role in coordinating the organism's stress response, including the activity of the hypothalamic-pituitary-adrenocortical axis. The molecular underpinnings of CRH-dependent signal transduction mechanisms in the anterior pituitary have not yet been revealed in detail. In order to dissect the signal transduction cascades activated by CRH receptor type 1, a comparative proteome approach was performed in vitro utilizing murine corticotroph AtT-20 cells. Alterations in protein expression and posttranslational modification in response to CRH stimulation were studied by 2D gel electrophoresis. Selected candidates were analyzed by immunoblotting and quantitative real-time PCR. The differential analyses revealed proteins regulated or modified related to diverse cellular processes. Amongst others we identified alterations in PRKAR1A, the regulatory subunit of protein kinase A; in PGK1 and PGAM1, key regulators of glycolysis; and in proteins involved in proteasome-mediated proteolysis, PSMC2 and PSMA3. These results offer novel entry points to molecular mechanisms underlying stress responses elicited via the hypothalamic-pituitary-adrenocortical axis.

Original language	English
Pages (from-to)	1-10
Number of pages	10
Journal	Molecular and Cellular Endocrinology
Volume	292
Issue number	1-2
DOIs	https://doi.org/10.1016/j.mce.2008.05.017
State	Published - 24 Sep 2008
Externally published	Yes

Keywords

AtT-20 cells
Corticotropin-releasing hormone
Corticotropin-releasing hormone receptor type 1
Hypothalamic-pituitary-adrenocortical axis
Proteome analysis

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10.1016/j.mce.2008.05.017

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title = "CRHR1-dependent effects on protein expression and posttranslational modification in AtT-20 cells",

abstract = "Corticotropin-releasing hormone (CRH) plays a major role in coordinating the organism's stress response, including the activity of the hypothalamic-pituitary-adrenocortical axis. The molecular underpinnings of CRH-dependent signal transduction mechanisms in the anterior pituitary have not yet been revealed in detail. In order to dissect the signal transduction cascades activated by CRH receptor type 1, a comparative proteome approach was performed in vitro utilizing murine corticotroph AtT-20 cells. Alterations in protein expression and posttranslational modification in response to CRH stimulation were studied by 2D gel electrophoresis. Selected candidates were analyzed by immunoblotting and quantitative real-time PCR. The differential analyses revealed proteins regulated or modified related to diverse cellular processes. Amongst others we identified alterations in PRKAR1A, the regulatory subunit of protein kinase A; in PGK1 and PGAM1, key regulators of glycolysis; and in proteins involved in proteasome-mediated proteolysis, PSMC2 and PSMA3. These results offer novel entry points to molecular mechanisms underlying stress responses elicited via the hypothalamic-pituitary-adrenocortical axis.",

keywords = "AtT-20 cells, Corticotropin-releasing hormone, Corticotropin-releasing hormone receptor type 1, Hypothalamic-pituitary-adrenocortical axis, Proteome analysis",

author = "Kronsbein, \{Helena C.\} and Jastorff, \{Archana M.\} and Giuseppina Maccarrone and G{\"u}nter Stalla and Wolfgang Wurst and Florian Holsboer and Turck, \{Christoph W.\} and Deussing, \{Jan M.\}",

note = "Funding Information: We thank Johanna Stalla for experimental support with the ACTH radioimmunoassays. This work was partially supported by the Bundesministerium f{\"u}r Bildung und Forschung within the framework of the NGFN2 (01GS0481) and by the Fonds der Chemi-schen Industrie.",

year = "2008",

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doi = "10.1016/j.mce.2008.05.017",

language = "English",

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journal = "Molecular and Cellular Endocrinology",

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T1 - CRHR1-dependent effects on protein expression and posttranslational modification in AtT-20 cells

AU - Kronsbein, Helena C.

AU - Jastorff, Archana M.

AU - Maccarrone, Giuseppina

AU - Stalla, Günter

AU - Wurst, Wolfgang

AU - Holsboer, Florian

AU - Turck, Christoph W.

AU - Deussing, Jan M.

N1 - Funding Information: We thank Johanna Stalla for experimental support with the ACTH radioimmunoassays. This work was partially supported by the Bundesministerium für Bildung und Forschung within the framework of the NGFN2 (01GS0481) and by the Fonds der Chemi-schen Industrie.

PY - 2008/9/24

Y1 - 2008/9/24

N2 - Corticotropin-releasing hormone (CRH) plays a major role in coordinating the organism's stress response, including the activity of the hypothalamic-pituitary-adrenocortical axis. The molecular underpinnings of CRH-dependent signal transduction mechanisms in the anterior pituitary have not yet been revealed in detail. In order to dissect the signal transduction cascades activated by CRH receptor type 1, a comparative proteome approach was performed in vitro utilizing murine corticotroph AtT-20 cells. Alterations in protein expression and posttranslational modification in response to CRH stimulation were studied by 2D gel electrophoresis. Selected candidates were analyzed by immunoblotting and quantitative real-time PCR. The differential analyses revealed proteins regulated or modified related to diverse cellular processes. Amongst others we identified alterations in PRKAR1A, the regulatory subunit of protein kinase A; in PGK1 and PGAM1, key regulators of glycolysis; and in proteins involved in proteasome-mediated proteolysis, PSMC2 and PSMA3. These results offer novel entry points to molecular mechanisms underlying stress responses elicited via the hypothalamic-pituitary-adrenocortical axis.

AB - Corticotropin-releasing hormone (CRH) plays a major role in coordinating the organism's stress response, including the activity of the hypothalamic-pituitary-adrenocortical axis. The molecular underpinnings of CRH-dependent signal transduction mechanisms in the anterior pituitary have not yet been revealed in detail. In order to dissect the signal transduction cascades activated by CRH receptor type 1, a comparative proteome approach was performed in vitro utilizing murine corticotroph AtT-20 cells. Alterations in protein expression and posttranslational modification in response to CRH stimulation were studied by 2D gel electrophoresis. Selected candidates were analyzed by immunoblotting and quantitative real-time PCR. The differential analyses revealed proteins regulated or modified related to diverse cellular processes. Amongst others we identified alterations in PRKAR1A, the regulatory subunit of protein kinase A; in PGK1 and PGAM1, key regulators of glycolysis; and in proteins involved in proteasome-mediated proteolysis, PSMC2 and PSMA3. These results offer novel entry points to molecular mechanisms underlying stress responses elicited via the hypothalamic-pituitary-adrenocortical axis.

KW - AtT-20 cells

KW - Corticotropin-releasing hormone

KW - Corticotropin-releasing hormone receptor type 1

KW - Hypothalamic-pituitary-adrenocortical axis

KW - Proteome analysis

UR - https://www.scopus.com/pages/publications/49349100570

U2 - 10.1016/j.mce.2008.05.017

DO - 10.1016/j.mce.2008.05.017

M3 - Article

C2 - 18582531

AN - SCOPUS:49349100570

SN - 0303-7207

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JO - Molecular and Cellular Endocrinology

JF - Molecular and Cellular Endocrinology

IS - 1-2

ER -

CRHR1-dependent effects on protein expression and posttranslational modification in AtT-20 cells

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