Figure 2.

KIR CL6 CYT-SHP-2 binding regulated by LCK. Yeast were transformed with the indicated TetR-CL6 CYT, LexA-LCK or TetR-LCK, and TA-SHP-2 hybrids. A - symbol indicates that yeast were transformed with the appropriate DNA binding (TetR or LexA) or TA components of hybrids only. In A and C yeast growth was assessed after 2 days (TetR-LCK) or 3 days (LexA-LCK) culture on Gal+ Raff+/LHWUra dropout plates. In B yeast were lysed, and phosphotyrosine-containing proteins were detected by Western blotting with an anti-phosphotyrosine mAb (Top). Blots were stripped and reprobed with an anti-TetR antiserum (Middle) and an anti-hemagglutinin mAb (Bottom) to reveal the position of hybrids (a hemagglutinin epitope tag is engineered between the TA and SHP-2). Note that the Y282F mutation of CL6 CYT increases the electrophoretic mobility of TetR-CL6 CYT.