Figure 4. Small molecule inhibitors, INHs, inhibit tumor growth by disrupting interaction between RB-interacting protein, Hec1, and Nek2.
(A) A schematic representation of a reverse yeast two-hybrid system for high-throughput screening for Hec1/Nek2 inhibitors. TetR fused with the C-terminal binding region of Hec1 (TetR-Hec1) was constitutively expressed; activation domain of GAL1 fused with Nek2 (AD-Nek2) was under the control of GAL1-inducible promoter. If an inhibitor abolishes the Hec1/Nek2 interaction, 5-FOA will not be metabolized, thus permitting yeast growth; otherwise, 5-FOA will be hydrolyzed into toxic metabolites and inhibit yeast growth. (B) Structures of two candidate INH compounds that promote yeast growth, i.e., disrupt Hec1/Nek2 interaction. (Figure adapted from (137))
