Chemical genetics approach reveals the role of Raf kinase inhibitor protein in cell migration.

摘要

Using chemical genetics approach, over 10,000 small-molecule compounds were screened for cell migration inhibitors. Totally 14 compounds with inhibitory activity toward cell migration were discovered.;Following one of the active compounds, locostatin, Raf kinase inhibitor protein (RKIP) was identified as the relevant target of locostatin. This protein shows a positive role in controlling cell migration: its overexpression promotes cell migration and cell-substratum adhesion, causes dramatic cell morphological change to a more motile phenotype; Knockdown of the protein inhibits cell migration as well as cell-substratum adhesion.;G-protein coupled receptor kinase 2 (GRK2) was proved to be involved in RKIP-mediated cell migration. RKIP associates and activates GRK2 both in vitro and in vivo. This interaction is inhibited by locostatin. GRK2 activity is required for cell migration. Knockdown of GRK2 inhibits cell migration and cell-substratum adhesion. Knockdown of GRK2 also reverses the cell morphological change caused by RKIP overexpression.;These findings revealed the positive role of RKIP in cell migration and GRK2's involvement in RKIP-mediated cell migration for the first time, suggested new mechanisms in controlling cell migration, which are of both scientific and clinical importance.