engleski

Assaying Rho GTPase–dependent processes in Dictyostelium discoideum

The model organism D. discoideum is well-suited to investigate basic questions of molecular and cell biology, particularly those related to the structure, regulation and dynamics of the cytoskeleton, signal transduction, cell-cell adhesion and development. D. discoideum cells make use of Rho-regulated signaling pathways to reorganize the actin cytoskeleton during chemotaxis, endocytosis and cytokinesis. In this organism the Rho family encompasses 20 members, several belonging to the Rac subfamily, but there are no representatives of the Cdc42 and Rho subfamilies. Here we present protocols suitable for monitoring the actin polymerization response and the activation of Rac upon stimulation of aggregation competent cells with the chemoattractant cAMP, and for monitoring the localization and dynamics of Rac activity in live cells.

Actin polymerization ; cAMP, chemotaxis ; Dictyostelium ; fluorescent probes ; FRET ; GTPase-binding domain ; pull-down assay ; Rac

Rad je financiran s istraživačkog projekta Hrvatske zaklade za znanost (šifra 4753): Oscilatorna dinamika citoskeleta (OSCITON)