Table 2.

Analysis of the interaction between DOK proteins and the insulin receptor in the yeast two-hybrid systema

Plasmidβ-Galactosidase activity when cotransformed with plasmid:
IR-CIR-CKRpSH2-Tyro3pSH2-AxlAGP4
pGD-DOKL++++
pGD-DOKLR209A +/−
pGD-p62d o k ++++
pGD-p62d o kR207A+/−
pGD-Grb2++++++
pGD-Vav p95++++++
  • a Plasmids in the heading contained different DNA fragments in frame with the LexA DNA-binding domain. Plasmids in the left column contained different DNA fragments in frame with the Gal4 activation domain. Yeast strain CTY 10-5d was transformed with various pairs of plasmids, followed by an X-Gal staining assay. The strength of interaction was indicated by the timing and intensity of blue color. +++, dark blue visible in 1 h; +, blue visible in 6 h; +/−, pale blue visible overnight; −, white overnight. IR-C represents the intracellular region of the insulin receptor, and IR-CKR represents the intracellular region of the insulin receptor with the defective kinase activity.