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PMID: 25614622 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Phosphatidylinositol 3-kinase class II α-isoform PI3K-C2α is required for transforming growth factor β-induced Smad signaling in endothelial cells.

The Journal of biological chemistry ·Vol. 290 ·No. 10 ·2015-03-06 ·Pages 6086-105

Aki S, Yoshioka K, Okamoto Y, Takuwa N, Takuwa Y

Abstract

We have recently demonstrated that the PI3K class II-α isoform (PI3K-C2α), which generates phosphatidylinositol 3-phosphate and phosphatidylinositol 3,4-bisphosphates, plays crucial roles in angiogenesis, by analyzing PI3K-C2α knock-out mice. The PI3K-C2α actions are mediated at least in part through its participation in the internalization of VEGF receptor-2 and sphingosine-1-phosphate receptor S1P1 and thereby their signaling on endosomes. TGFβ, which is also an essential angiogenic factor, signals via the serine/threonine kinase receptor complex to induce phosphorylation of Smad2 and Smad3 (Smad2/3). SARA (Smad anchor for receptor activation) protein, which is localized in early endosomes through its FYVE domain, is required for Smad2/3 signaling. In the present study, we showed that PI3K-C2α knockdown nearly completely abolished TGFβ1-induced phosphorylation and nuclear translocation of Smad2/3 in vascular endothelial cells (ECs). PI3K-C2α was necessary for TGFβ-induced increase in phosphatidylinositol 3,4-bisphosphates in the plasma membrane and TGFβ receptor internalization into the SARA-containing early endosomes, but not for phosphatidylinositol 3-phosphate enrichment or localization of SARA in the early endosomes. PI3K-C2α was also required for TGFβ receptor-mediated formation of SARA-Smad2/3 complex. Inhibition of dynamin, which is required for the clathrin-dependent receptor endocytosis, suppressed both TGFβ receptor internalization and Smad2/3 phosphorylation. TGFβ1 stimulated Smad-dependent VEGF-A expression, VEGF receptor-mediated EC migration, and capillary-like tube formation, which were all abolished by either PI3K-C2α knockdown or a dynamin inhibitor. Finally, TGFβ1-induced microvessel formation in Matrigel plugs was greatly attenuated in EC-specific PI3K-C2α-deleted mice. These observations indicate that PI3K-C2α plays the pivotal role in TGFβ receptor endocytosis and thereby Smad2/3 signaling, participating in angiogenic actions of TGFβ.

Keywords
Endosome Endothelial Cell PI3K-C2alpha Phosphatidylinositol Kinase (PI Kinase) Receptor Endocytosis SARA SMAD Transcription Factor TGF-B Receptor Transforming Growth Factor Beta (TGF-B) Vascular Endothelial Growth Factor (VEGF)
MeSH Terms
Animals Endocytosis/genetics Endothelial Cells/metabolism Gene Expression Regulation Human Umbilical Vein Endothelial Cells Humans Intracellular Signaling Peptides and Proteins/biosynthesis,genetics Mice Mice, Knockout Phosphatidylinositol 3-Kinases/genetics Serine Endopeptidases/biosynthesis,genetics Signal Transduction Transforming Growth Factor beta1/genetics,metabolism Vascular Endothelial Growth Factor A/genetics,metabolism
Chemicals
Intracellular Signaling Peptides and Proteins Transforming Growth Factor beta1 Vascular Endothelial Growth Factor A Phosphatidylinositol 3-Kinases PIK3C2A protein, human ZFYVE16 protein, human Serine Endopeptidases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Aki Sho
From the Department of Physiology, Kanazawa University School of Medicine, Kanazawa, Ishikawa 920-8640, Japan and.
Yoshioka Kazuaki
From the Department of Physiology, Kanazawa University School of Medicine, Kanazawa, Ishikawa 920-8640, Japan and.
Okamoto Yasuo
From the Department of Physiology, Kanazawa University School of Medicine, Kanazawa, Ishikawa 920-8640, Japan and.
Takuwa Noriko
From the Department of Physiology, Kanazawa University School of Medicine, Kanazawa, Ishikawa 920-8640, Japan and the Department of Health and Medical Sciences, Ishikawa Prefectural Nursing University, Kahoku, Ishikawa 929-1210, Japan.
Takuwa Yoh
From the Department of Physiology, Kanazawa University School of Medicine, Kanazawa, Ishikawa 920-8640, Japan and ytakuwa@med.kanazawa-u.ac.jp.
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M114.601484
Email
ytakuwa@med.kanazawa-u.ac.jp
Published
2015-03-06
Epub
2015-00-22
Pages
6086-105
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC4358250
Subset
IM
External Links
PubMed source DOI: 10.1074/jbc.M114.601484 PMC Full Text View journal details
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