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PMID: 18606851 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Sequential signals toward podosome formation in NIH-src cells.

The Journal of cell biology ·Vol. 182 ·No. 1 ·2008-07-14 ·Pages 157-69

Oikawa T, Itoh T, Takenawa T

Abstract

Podosomes (also termed invadopodia in cancer cells) are actin-rich adhesion structures with matrix degradation activity that develop in various cell types. Despite their significant physiological importance, the molecular mechanism of podosome formation is largely unknown. In this study, we investigated the molecular mechanisms of podosome formation. The expression of various phosphoinositide-binding domains revealed that the podosomes in Src-transformed NIH3T3 (NIH-src) cells are enriched with PtdIns(3,4)P2, suggesting an important role of this phosphoinositide in podosome formation. Live-cell imaging analysis revealed that Src-expression stimulated podosome formation at focal adhesions of NIH3T3 cells after PtdIns(3,4)P2 accumulation. The adaptor protein Tks5/FISH, which is essential for podosome formation, was found to form a complex with Grb2 at adhesion sites in an Src-dependent manner. Further, it was found that N-WASP bound all SH3 domains of Tks5/FISH, which facilitated circular podosome formation. These results indicate that augmentation of the N-WASP-Arp2/3 signal was accomplished on the platform of Tks5/FISH-Grb2 complex at focal adhesions, which is stabilized by PtdIns(3,4)P2.

MeSH Terms
Actins/metabolism Animals Cell Line, Transformed Cell Surface Extensions/metabolism Cell Survival Enzyme Activation Focal Adhesions/enzymology GRB2 Adaptor Protein/metabolism Mice Mutation/genetics NIH 3T3 Cells Phosphate-Binding Proteins Phosphatidylinositol Phosphates/metabolism Phosphoproteins/metabolism Protein Transport Sequence Homology Signal Transduction Wiskott-Aldrich Syndrome Protein, Neuronal/chemistry src-Family Kinases/metabolism
Chemicals
Actins Fish protein, mouse GRB2 Adaptor Protein Phosphate-Binding Proteins Phosphatidylinositol Phosphates Phosphoproteins Wiskott-Aldrich Syndrome Protein, Neuronal phosphatidylinositol 3,4-diphosphate src-Family Kinases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Oikawa Tsukasa
Division of Lipid Biochemistry, Department of Biochemistry and Molecular Biology, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe, Hyogo 650-0017, Japan.
Itoh Toshiki
Takenawa Tadaomi
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
1540-8140
DOI
10.1083/jcb.200801042
Published
2008-07-14
Epub
2008-00-07
Pages
157-69
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2447888
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.200801042 PMC Full Text View journal details
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