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PMID: 11094076 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Inhibitory regulation of Rac activation, membrane ruffling, and cell migration by the G protein-coupled sphingosine-1-phosphate receptor EDG5 but not EDG1 or EDG3.

Molecular and cellular biology ·Vol. 20 ·No. 24 ·2000-12-00 ·Pages 9247-61

Okamoto H, Takuwa N, Yokomizo T, Sugimoto N, Sakurada S, Shigematsu H, Takuwa Y

Abstract

Sphingosine-1-phosphate (S1P) is a bioactive lysophospholipid that induces a variety of biological responses in diverse cell types. Many, if not all, of these responses are mediated by members of the EDG (endothelial differentiation gene) family G protein-coupled receptors EDG1, EDG3, and EDG5 (AGR16). Among prominent activities of S1P is the regulation of cell motility; S1P stimulates or inhibits cell motility depending on cell types. In the present study, we provide evidence for EDG subtype-specific, contrasting regulation of cell motility and cellular Rac activity. In CHO cells expressing EDG1 or EDG3 (EDG1 cells or EDG3 cells, respectively) S1P as well as insulin-like growth factor I (IGF I) induced chemotaxis and membrane ruffling in phosphoinositide (PI) 3-kinase- and Rac-dependent manners. Both S1P and IGF I induced a biphasic increase in the amount of the GTP-bound active form of Rac. In CHO cells expressing EDG5 (EDG5 cells), IGF I similarly stimulated cell migration; however, in contrast to what was found for EDG1 and EDG3 cells, S1P did not stimulate migration but totally abolished IGF I-directed chemotaxis and membrane ruffling, in a manner dependent on a concentration gradient of S1P. In EDG5 cells, S1P stimulated PI 3-kinase activity as it did in EDG1 cells but inhibited the basal Rac activity and totally abolished IGF I-induced Rac activation, which involved stimulation of Rac-GTPase-activating protein activity rather than inhibition of Rac-guanine nucleotide exchange activity. S1P induced comparable increases in the amounts of GTP-RhoA in EDG3 and EDG5 cells. Neither S1P nor IGF I increased the amount of GTP-bound Cdc42. However, expression of N(17)-Cdc42, but not N(19)-RhoA, suppressed S1P- and IGF I-directed chemotaxis, suggesting a requirement for basal Cdc42 activity for chemotaxis. Taken together, the present results demonstrate that EDG5 is the first example of a hitherto-unrecognized type of receptors that negatively regulate Rac activity, thereby inhibiting cell migration and membrane ruffling.

MeSH Terms
3T3 Cells Animals CHO Cells Cell Membrane/metabolism,ultrastructure Chemotaxis/drug effects Cricetinae DNA-Binding Proteins/metabolism Humans I-kappa B Proteins Immediate-Early Proteins/metabolism Insulin-Like Growth Factor I/pharmacology Lysophospholipids Mice NF-KappaB Inhibitor alpha Phosphatidylinositol 3-Kinases/metabolism Protein Isoforms/metabolism Protein Kinase Inhibitors Protein Serine-Threonine Kinases/metabolism Receptors, Cell Surface/metabolism Receptors, G-Protein-Coupled Receptors, Lysophospholipid Recombinant Proteins/genetics,metabolism Signal Transduction Sphingosine/analogs & derivatives,metabolism,pharmacology Stress Fibers/metabolism Transfection cdc42 GTP-Binding Protein/metabolism p21-Activated Kinases rac GTP-Binding Proteins/antagonists & inhibitors,metabolism rho GTP-Binding Proteins/metabolism
Chemicals
DNA-Binding Proteins I-kappa B Proteins Immediate-Early Proteins Lysophospholipids NFKBIA protein, human Nfkbia protein, mouse Protein Isoforms Protein Kinase Inhibitors Receptors, Cell Surface Receptors, G-Protein-Coupled Receptors, Lysophospholipid Recombinant Proteins NF-KappaB Inhibitor alpha sphingosine 1-phosphate Insulin-Like Growth Factor I Protein Serine-Threonine Kinases p21-Activated Kinases cdc42 GTP-Binding Protein rac GTP-Binding Proteins rho GTP-Binding Proteins Sphingosine
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Okamoto H
Department of Physiology, Kanazawa University School of Medicine, Kanazawa, Japan.
Takuwa N
Yokomizo T
Sugimoto N
Sakurada S
Shigematsu H
Takuwa Y
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.20.24.9247-9261.2000
Published
2000-12-00
Pages
9247-61
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC102182
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.20.24.9247-9261.2000 PMC Full Text View journal details
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