Exp Mol Med.  2008 Jun;40(3):261-270. 10.3858/emm.2008.40.3.261.

CD98 activation increases surface expression and clusteringof beta 1 integrins in MCF-7 cells through FAK/Src- and cytoskeleton-independent mechanisms

• Kim SM
• Hahn JH

• KMID: 756327
• DOI: http://doi.org/10.3858/emm.2008.40.3.261

Abstract

CD98, a disulfide-linked 125-kDa heterodimeric type II transmembrane glycoprotein, regulates beta 1 integrin- mediated cell adhesion. However, the molecular mechanisms underlying CD98-mediated activation of beta 1 integrin are presently unclear. In this study, the effects of CD98 signaling on the expression and clustering of beta 1 integrin were investigated. Activation of CD98 augmented surface expression of beta 1 integrin on MCF-7 cells. Cross-linking CD98 induced clustering of beta 1 integrins. Inhibition of phosphorylation of focal adhesion kimase (FAK) by PP2, an inhibitor of Src family kinase, reduced cell-extracellular matrix adhesion, but not surface expression and clustering of beta1 integrin on MCF-7 cells. This result was confirmed by over-expression of dominant negative forms of FAK. In addition, phalloidin or cytochalasin D inhibited CD98-mediated induction of cell-ECM adhesion, but not surface expression and clustering of b1 integrins. The inhibitory effects of PP2, cytochalasin D or phalloidin on CD98-stimulated cell adhesion were diminished by pretreatment of cells with Mn2+, which is shown to induce conformational change of integrins. These results provide the first evidence that CD98 activation increases not only beta1 integrin affinity but also its surface expression and clustering and the latter is independent of FAK/Src and cytoskeleton.

Keyword

actins; antigens, CD98; cytoskeleton; focal adhesion kinase; integrin beta 1

MeSH Terms

Antigens, CD29/*biosynthesis/genetics
Antigens, CD98/agonists/*metabolism
Cell Line, Tumor
Cytochalasin D/pharmacology
Cytoskeleton/drug effects/enzymology
Focal Adhesion Kinase 2/genetics/*metabolism
Focal Adhesions/drug effects/enzymology
Humans
Microscopy, Confocal
Multiprotein Complexes/*biosynthesis/genetics
Mutant Proteins/genetics/metabolism
Phalloidine/pharmacology
Phosphorylation/drug effects
Protein Binding
Pyrimidines/pharmacology
Signal Transduction/physiology
Transfection