title: GILZ mediates the antiproliferative activity of glucocorticoids by negative regulation of Ras signaling.
abstract: Tsc22d3 coding for glucocorticoid-induced leucine zipper (GILZ) was initially identified as a dexamethasone-responsive gene involved in the control of T lymphocyte activation and apoptosis. However, the physiological role of this molecule and its function in the biological activity of glucocorticoids (GCs) has not been clarified. Here, we demonstrate that GILZ interacts directly with Ras in vitro and in vivo as shown by GILZ and Ras coimmunoprecipitation and colocalization upon PMA activation in primary mouse spleen T lymphocytes and thymus cells. The analysis of GILZ mutants showed that they bound Ras through the tuberous sclerosis complex box (TSC) and, depending on the Ras activation level, formed a trimeric complex with Ras and Raf, which we previously identified as a GILZ binder. As a consequence of these interactions, GILZ diminished the activation of Ras and Raf downstream targets including ERK1/2, AKT/PKB serine/threonine kinase, and retinoblastoma (Rb) phosphorylation and cyclin D1 expression, leading to inhibition of Ras- and Raf-dependent cell proliferation and Ras-induced NIH-3T3 transformation. GILZ silencing resulted in an increase in concanavalin A-induced T cell proliferation and, most notably, inhibition of dexamethasone antiproliferative effects. Together, these findings indicate that GILZ serves as a negative regulator of Ras- and Raf-induced proliferation and is an important mediator of the antiproliferative effect of GCs.
authors.affiliation: Department of Clinical and Experimental Medicine, Section of Pharmacology, University of Perugia, Perugia, Italy.
authors.forename: Emira
authors.initials: E
authors.lastname: Ayroldi
authors.forename: Ornella
authors.initials: O
authors.lastname: Zollo
authors.forename: Alessandra
authors.initials: A
authors.lastname: Bastianelli
authors.forename: Cristina
authors.initials: C
authors.lastname: Marchetti
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Agostini
authors.forename: Rosa
authors.initials: R
authors.lastname: Di Virgilio
authors.forename: Carlo
authors.initials: C
authors.lastname: Riccardi
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D003907:Dexamethasone / Q000494:pharmacology*; D005938:Glucocorticoids / Q000494:pharmacology; D066298:In Vitro Techniques; D051379:Mice; D008809:Mice, Inbred C3H; D009154:Mutation; D041681:NIH 3T3 Cells; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D018631:ras Proteins / Q000378:metabolism*
chemical_list: C498608:Dsip1 protein, mouse; D005938:Glucocorticoids; D011994:Recombinant Proteins; D014157:Transcription Factors; D003907:Dexamethasone; D018631:ras Proteins