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.
authors.affiliation: Max Planck Institute of Psychiatry, Kraepelinstrasse 2-10, D-80804 Munich, Germany.
authors.forename: Helena C
authors.initials: HC
authors.lastname: Kronsbein
authors.forename: Archana M
authors.initials: AM
authors.lastname: Jastorff
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Maccarrone
authors.forename: Günter
authors.initials: G
authors.lastname: Stalla
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Wurst
authors.forename: Florian
authors.initials: F
authors.lastname: Holsboer
authors.forename: Christoph W
authors.initials: CW
authors.lastname: Turck
authors.forename: Jan M
authors.initials: JM
authors.lastname: Deussing
mesh_terms: D000324:Adrenocorticotropic Hormone / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D003346:Corticotropin-Releasing Hormone / Q000494:pharmacology; D054756:Cyclic AMP-Dependent Protein Kinase RIalpha Subunit / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D013058:Mass Spectrometry; D051379:Mice; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D011506:Proteins / Q000378:metabolism*; D018019:Receptors, Corticotropin-Releasing Hormone / Q000378:metabolism*; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D000099052:CRF Receptor, Type 1
chemical_list: D054756:Cyclic AMP-Dependent Protein Kinase RIalpha Subunit; C517835:Prkar1a protein, mouse; D011506:Proteins; D018019:Receptors, Corticotropin-Releasing Hormone; D000099052:CRF Receptor, Type 1; D000324:Adrenocorticotropic Hormone; D003346:Corticotropin-Releasing Hormone; D046988:Proteasome Endopeptidase Complex