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 glycolipidenriched 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.

Ganglioside GD3 Enhances Invasiveness of Gliomas by Forming a Complex with Platelet-derived Growth Factor Receptor α and Yes Kinase

“…Kleber et al (50) reported that Yes and the p85 subunit of PI3K are recruited to CD95 by stimulation with CD95 ligand, resulting in an intensive invasion phenotype in glioblastoma multiforme. Our group also reported that GD3 is essential for the functional activation of Yes in human melanoma cells (51). Moreover, the FAK/Src family protein complex activates paxillin via phosphorylation at tyrosine residues 31 and 118 (52).…”

Neogenin, Defined as a GD3-associated Molecule by Enzyme-mediated Activation of Radical Sources, Confers Malignant Properties via Intracytoplasmic Domain in Melanoma Cells

“…The effects of anti-GD3 antibody therapy in melanomas (16) and various aspects of its roles in melanomas have been also reported (17)(18)(19). These results suggest that GD3 plays important roles in the enhancement of malignant properties of melanomas (20,21) by regulating glycolipid-enriched microdomain (GEM) 2 /rafts (22).…”

Glycolipid GD3 and GD3 synthase are key drivers for glioblastoma stem cells and tumorigenicity

“…The functional role of GD3 in GSCs is not clear. Recent studies suggest that gangliosides are widely expressed on tumor cells (36) and disialyl gangliosides enhance tumor phenotype with multiple modalities (37). These reports demonstrate that GD3 promotes cell growth and invasion through phosphorylation of p130Cas, paxillin, and focal adhesion kinase (FAK) in melanoma cells (38).…”

“…These reports demonstrate that GD3 promotes cell growth and invasion through phosphorylation of p130Cas, paxillin, and focal adhesion kinase (FAK) in melanoma cells (38). Furthermore, the Src family kinase Yes was found to link with p130Cas or FAK as an activated form to promote malignant properties of human melanoma cells expressing GD3 (37,39). It was also revealed that gangliosides, including GD1a, GD1b, GD3, and GM3, may facilitate tumor cell escape from the immune system within the tumor microenvironment (40).…”