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PMID: 23192342 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Essential role of class II phosphatidylinositol-3-kinase-C2α in sphingosine 1-phosphate receptor-1-mediated signaling and migration in endothelial cells.

The Journal of biological chemistry ·Vol. 288 ·No. 4 ·2013-01-25 ·Pages 2325-39

Biswas K, Yoshioka K, Asanuma K, Okamoto Y, Takuwa N, Sasaki T, Takuwa Y

Abstract

The phosphatidylinositol (PtdIns) 3-kinase (PI3K) family regulates diverse cellular processes, including cell proliferation, migration, and vesicular trafficking, through catalyzing 3'-phosphorylation of phosphoinositides. In contrast to class I PI3Ks, including p110α and p110β, functional roles of class II PI3Ks, comprising PI3K-C2α, PI3K-C2β, and PI3K-C2γ, are little understood. The lysophospholipid mediator sphingosine 1-phosphate (S1P) plays the important roles in regulating vascular functions, including vascular formation and barrier integrity, via the G-protein-coupled receptors S1P(1-3). We studied the roles of PI3K-C2α in S1P-induced endothelial cell (EC) migration and tube formation. S1P stimulated cell migration and activation of Akt, ERK, and Rac1, the latter of which acts as a signaling molecule essential for cell migration and tube formation, via S1P(1) in ECs. Knockdown of either PI3K-C2α or class I p110β markedly inhibited S1P-induced migration, lamellipodium formation, and tube formation, whereas that of p110α or Vps34 did not. Only p110β was necessary for S1P-iduced Akt activation, but both PI3K-C2α and p110β were required for Rac1 activation. FRET imaging showed that S1P induced Rac1 activation in both the plasma membrane and PtdIns 3-phosphate (PtdIns(3)P)-enriched endosomes. Knockdown of PI3K-C2α but not p110β markedly reduced PtdIns(3)P-enriched endosomes and suppressed endosomal Rac1 activation. Also, knockdown of PI3K-C2α but not p110β suppressed S1P-induced S1P(1) internalization into PtdIns(3)P-enriched endosomes. Finally, pharmacological inhibition of endocytosis suppressed S1P-induced S1P(1) internalization, Rac1 activation, migration, and tube formation. These observations indicate that PI3K-C2α plays the crucial role in S1P(1) internalization into the intracellular vesicular compartment, Rac1 activation on endosomes, and thereby migration through regulating vesicular trafficking in ECs.

MeSH Terms
Cell Movement Cells, Cultured Class II Phosphatidylinositol 3-Kinases/genetics,physiology Endocytosis Endosomes/metabolism Endothelial Cells/cytology Fluorescence Resonance Energy Transfer Gene Expression Regulation, Enzymologic Human Umbilical Vein Endothelial Cells Humans Lysophospholipids/metabolism RNA, Small Interfering/metabolism Receptors, Lysosphingolipid/genetics Signal Transduction Transfection rac GTP-Binding Proteins/metabolism
Chemicals
Lysophospholipids RNA, Small Interfering Receptors, Lysosphingolipid Class II Phosphatidylinositol 3-Kinases rac GTP-Binding Proteins lysophosphatidic acid
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Biswas Kuntal
Department of Physiology, Kanazawa University School of Medicine, 13-1 Takara-machi, Kanazawa, Ishikawa 920-8640, Japan.
Yoshioka Kazuaki
Asanuma Ken
Okamoto Yasuo
Takuwa Noriko
Sasaki Takehiko
Takuwa Yoh
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M112.409656
Published
2013-01-25
Epub
2012-00-28
Pages
2325-39
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC3554904
Subset
IM
External Links
PubMed source DOI: 10.1074/jbc.M112.409656 PMC Full Text View journal details
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