Assaying Rho GTPase–dependent processes in Dictyostelium discoideum

Marinovic, Maja; Xiong, Huajiang; Rivero Crespo, Francisco; Weber, Igor

Abstract

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.

Citation

Marinovic, M., Xiong, H., Rivero Crespo, F., & Weber, I. (2018). Assaying Rho GTPase–dependent processes in Dictyostelium discoideum. In F. Rivero (Ed.), Rho GTPases: Methods and Protocols (371-392). https://doi.org/10.1007/978-1-4939-8612-5_25

• Acceptance Date: May 11, 2018
• Online Publication Date: Jul 31, 2018
• Publication Date: Jul 31, 2018
• Deposit Date: Jul 11, 2018
• Publicly Available Date: Aug 1, 2019
• Peer Reviewed: Peer Reviewed
• Pages: 371-392
• Series Title: Methods in Molecular Biology
• Series Number: 1821
• Series ISSN: 1940-6029
• Book Title: Rho GTPases: Methods and Protocols
• ISBN: 978-1-4939-8611-8
• DOI: https://doi.org/10.1007/978-1-4939-8612-5_25
• Keywords: Actin polymerization; cAMP; Chemotaxis; Dictyostelium; Fluorescent probes; FRET; GTPase-binding domain; Pull-down assay, Rac
• Public URL: https://hull-repository.worktribe.com/output/923295