Regulation of actin cytoskeleton by Rap1 binding to RacGEF1.
Rap1 is rapidly and transiently activated in response to chemoattractant stimulation and helps establish cell polarity by locally modulating cytoskeletons. Here, we investigated the mechanisms by which Rap1 controls actin cytoskeletal reorganization in Dictyostelium and found that Rap1 interacts with RacGEF1 in vitro and stimulates F-actin polymerization at the sites where Rap1 is activated upon chemoattractant stimulation. Live cell imaging using GFP-coronin, a reporter for F-actin, demonstrates that cells expressing constitutively active Rap1 (Rap1CA) exhibit a high level of F-actin uniformly distributed at the cortex including the posterior and lateral sides of the chemotaxing cell. Examination of the localization of a PH-domain containing PIP3 reporter, PhdA-GFP, and the activation of Akt/Pkb and other Ras proteins in Rap1CA cells reveals that activated Rap1 has no effect on the production of PIP3 or the activation of Akt/Pkb and Ras proteins in response to chemoattractant stimulation. Rac family proteins are crucial regulators in actin cytoskeletal reorganization. In vitro binding assay using truncated RacGEF1 proteins shows that Rap1 interacts with the DH domain of RacGEF1. Taken together, these results suggest that Rap1-mediated F-actin polymerization probably occurs through the Rac signaling pathway by directly binding to RacGEF1.
Department of Biology, College of Natural Sciences, Chosun University, Gwangju 501-759, Korea.
Hyemin
H
Mun
Taeck J
TJ
Jeon
D008841:Actin Cytoskeleton / Q000378:metabolism*; D002630:Chemotactic Factors / Q000502:physiology; D002633:Chemotaxis; D004023:Dictyostelium / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D004789:Enzyme Activation; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D007700:Kinetics; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D015800:Protozoan Proteins / Q000378:metabolism*; D015398:Signal Transduction; D020827:rap1 GTP-Binding Proteins / Q000378:metabolism*; D018631:ras Proteins / Q000378:metabolism
D002630:Chemotactic Factors; D020662:Guanine Nucleotide Exchange Factors; D015800:Protozoan Proteins; D019869:Phosphatidylinositol 3-Kinases; D020827:rap1 GTP-Binding Proteins; D018631:ras Proteins