title: Localization of nectin-free afadin at the leading edge and its involvement in directional cell movement induced by platelet-derived growth factor.
abstract: Afadin is an actin-filament-binding protein that binds to nectin, an immunoglobulin-like cell-cell adhesion molecule, and plays an important role in the formation of adherens junctions. Here, we show that afadin, which did not bind to nectin and was localized at the leading edge of moving cells, has another role: enhancement of the directional, but not random, cell movement. When NIH3T3 cells were stimulated with platelet-derived growth factor (PDGF), afadin colocalized with PDGF receptor, alphavbeta3 integrin and nectin-like molecule-5 at the leading edge and facilitated the formation of leading-edge structures and directional cell movement in the direction of PDGF stimulation. However, these phenotypes were markedly perturbed by knockdown of afadin, and were dependent on the binding of afadin to active Rap1. Binding of Rap1 to afadin was necessary for the recruitment of afadin and the tyrosine phosphatase SHP-2 to the leading edge. SHP-2 was previously reported to tightly regulate the activation of PDGF receptor and its downstream signaling pathway for the formation of the leading edge. These results indicate that afadin has a novel role in PDGF-induced directional cell movement, presumably in cooperation with active Rap1 and SHP-2.
authors.affiliation: Division of Molecular and Cellular Biology, Department of Biochemistry and Molecular Biology, Kobe University Graduate School of Medicine, Kobe 650-0017, Japan.
authors.forename: Muneaki
authors.initials: M
authors.lastname: Miyata
authors.forename: Hisakazu
authors.initials: H
authors.lastname: Ogita
authors.forename: Hitomi
authors.initials: H
authors.lastname: Komura
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Nakata
authors.forename: Ryoko
authors.initials: R
authors.lastname: Okamoto
authors.forename: Misa
authors.initials: M
authors.lastname: Ozaki
authors.forename: Takashi
authors.initials: T
authors.lastname: Majima
authors.forename: Naomi
authors.initials: N
authors.lastname: Matsuzawa
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kawano
authors.forename: Akihiro
authors.initials: A
authors.lastname: Minami
authors.forename: Masumi
authors.initials: M
authors.lastname: Waseda
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Fujita
authors.forename: Kiyohito
authors.initials: K
authors.lastname: Mizutani
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Rikitake
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Takai
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002417:Cattle; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects*; D047468:Immunoprecipitation; D051379:Mice; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D041681:NIH 3T3 Cells; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology*; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11 / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020827:rap1 GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; D009363:Neoplasm Proteins; D010982:Platelet-Derived Growth Factor; D011993:Recombinant Fusion Proteins; C476587:Taa1 protein, mouse; C108835:afadin; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D020827:rap1 GTP-Binding Proteins