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Investigating the Function of Coronin A in the Early Starvation Response of Dictyostelium discoideum by Aggregation Assays
Published on: June 18, 2016
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Assaying Rho GTPase-Dependent Processes in Dictyostelium discoideum
Maja Marinović1, Huajiang Xiong2, Francisco Rivero3
1Division of Molecular Biology, Ruđer Bošković Institute, Zagreb, Croatia.
Methods in Molecular Biology (Clifton, N.J.)
|August 1, 2018
Summary
Dictyostelium discoideum (D. discoideum) cells use Rac signaling to control actin cytoskeleton dynamics. This study presents methods to monitor Rac activation and actin polymerization in live D. discoideum cells during chemotaxis.
Area of Science:
- Molecular and Cell Biology
- Developmental Biology
- Biochemistry
Background:
- Dictyostelium discoideum (D. discoideum) is a model organism for studying cytoskeleton dynamics, signal transduction, and development.
- Rho-regulated signaling pathways are crucial for actin cytoskeleton reorganization in D. discoideum during processes like chemotaxis, endocytosis, and cytokinesis.
- The D. discoideum Rho family includes 20 members, with several Rac subfamily members but no Cdc42 or Rho subfamily representatives.
Purpose of the Study:
- To present protocols for monitoring actin polymerization response and Rac activation in D. discoideum.
- To enable the study of Rac activity dynamics in live cells upon chemoattractant stimulation.
Main Methods:
- Stimulation of aggregation-competent D. discoideum cells with the chemoattractant cyclic adenosine monophosphate (cAMP).
- Monitoring actin polymerization response.
- Monitoring Rac activation and localization in live cells.
Main Results:
- Protocols are presented for monitoring actin polymerization and Rac activation in response to cAMP.
- Methods allow for the observation of Rac activity localization and dynamics in real-time.
Conclusions:
- The developed protocols facilitate the investigation of Rac-mediated signaling in D. discoideum.
- These methods are valuable for understanding cytoskeleton regulation and cellular processes in this model organism.
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