the structural features of trask that mediate its anti-adhesive functions查斯克,调解anti-adhesive函数的结构特点.pdfVIP

The Structural Features of Trask That Mediate Its Anti- Adhesive Functions Danislav S. Spassov1,2, Deepika Ahuja1,2, Ching Hang Wong1,2, Mark M. Moasser1,2* 1 Department of Medicine, University of California San Francisco, San Francisco, California, United States of America, 2 Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, California, United States of America Abstract Trask/CDCP1 is a transmembrane protein with a large extracellular and small intracellular domains. The intracellular domain (ICD) undergoes tyrosine phosphorylation by Src kinases during anchorage loss and, when phosphorylated, Trask functions to inhibit cell adhesion. The extracellular domain (ECD) undergoes proteolytic cleavage by serine proteases, although the functional significance of this remains unknown. There is conflicting evidence regarding whether it functions to signal the phosphorylation of the ICD. To better define the structural determinants that mediate the anti-adhesive functions of Trask, we generated a series of deletion mutants of Trask and expressed them in tet-inducible cell models to define the structural elements involved in cell adhesion signaling. We find that the ECD is dispensable for the phosphorylation of the ICD or for the inhibition of cell adhesion. The anti-adhesive functions of Trask are entirely embodied within its ICD and are specifically due to tyrosine phosphorylation of the ICD as this function is completely lost in a phosphorylation-defective tyrosine- phenylalanine mutant. Both full length and cleaved ECDs are fully capable of phosphorylation and undergo phosphorylation during anchorage loss and cleavage is not an upstream signal for ICD phosphorylation. These data establish that the anti-adhesive functions of Trask are mediated entirely through it