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Assaying Rho GTPase–dependent processes in Dictyostelium discoideum

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

Authors

Maja Marinovic

Huajiang Xiong

Igor Weber



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. Methods in molecular biology, 1821, 371-392. https://doi.org/10.1007/978-1-4939-8612-5_25

Journal Article Type Article
Acceptance Date May 11, 2018
Online Publication Date Jul 31, 2018
Publication Date Jul 31, 2018
Deposit Date Jul 11, 2018
Publicly Available Date Mar 29, 2024
Print ISSN 1064-3745
Peer Reviewed Peer Reviewed
Volume 1821
Pages 371-392
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
Publisher URL https://link.springer.com/protocol/10.1007%2F978-1-4939-8612-5_25

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