title: Functional activation of Src family kinase yes protein is essential for the enhanced malignant properties of human melanoma cells expressing ganglioside GD3.
abstract: The possible roles of Src family kinases in the enhanced malignant properties of melanomas related to GD3 expression were analyzed. Among Src family kinases only Yes, not Fyn or Src, was functionally involved in the increased cell proliferation and invasion of GD3-expressing transfectant cells (GD3+). Yes was located upstream of p130Cas and paxillin and at an equivalent level to focal adhesion kinase. Yes underwent autophosphorylation even before serum treatment and showed stronger kinase activity in GD3+ cells than in GD3- cells following serum treatment. Coimmunoprecipitation experiments revealed that Yes bound to focal adhesion kinase or p130Cas more strongly in GD3+ cells than in GD3- cells. As a possible mechanism for the enhancing effects of GD3 on cellular phenotypes, it was shown that majority of Yes was localized in glycolipid-enriched microdomain/rafts in GD3+ cells even before serum treatment, whereas it was scarcely detected in glycolipid-enriched microdomain/rafts in GD3- cells. An in vitro kinase assay of Yes revealed that coexistence of GD3 with Yes in membranous environments enhances the kinase activity of GD3- cell-derived Yes toward enolase, p125, and Yes itself. Knockdown of GD3 synthase resulted in the alleviation of tumor phenotypes and reduced activation levels of Yes. Taken together, these results suggest a role of GD3 in the regulation of Src family kinases.
authors: Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065, USA.
authors: Kazunori
authors: K
authors: Hamamura
authors: Momoko
authors: M
authors: Tsuji
authors: Hiroshi
authors: H
authors: Hotta
authors: Yuki
authors: Y
authors: Ohkawa
authors: Masataka
authors: M
authors: Takahashi
authors: Hidenobu
authors: H
authors: Shibuya
authors: Hideyuki
authors: H
authors: Nakashima
authors: Yoshio
authors: Y
authors: Yamauchi
authors: Noboru
authors: N
authors: Hashimoto
authors: Hisashi
authors: H
authors: Hattori
authors: Minoru
authors: M
authors: Ueda
authors: Keiko
authors: K
authors: Furukawa
authors: Koichi
authors: K
authors: Furukawa
mesh_terms: D045744:Cell Line, Tumor; D050719:Crk-Associated Substrate Protein / Q000235:genetics / Q000378:metabolism; D004789:Enzyme Activation / Q000235:genetics; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D005732:Gangliosides / Q000096:biosynthesis* / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D006801:Humans; D008545:Melanoma / Q000235:genetics / Q000378:metabolism*; D021962:Membrane Microdomains / Q000235:genetics / Q000378:metabolism*; D010751:Phosphopyruvate Hydratase / Q000235:genetics / Q000378:metabolism; D051575:Proto-Oncogene Proteins c-yes / Q000235:genetics / Q000378:metabolism*; D012799:Sialyltransferases / Q000235:genetics / Q000378:metabolism
chemical_list: D050719:Crk-Associated Substrate Protein; D005732:Gangliosides; C026226:ganglioside, GD3; D012799:Sialyltransferases; C043662:alpha-N-acetylneuraminate alpha-2,8-sialyltransferase; D051416:Focal Adhesion Protein-Tyrosine Kinases; D051575:Proto-Oncogene Proteins c-yes; C489834:YES1 protein, human; D010751:Phosphopyruvate Hydratase