FIG. 4.

METT-10 binds DLC-1 in vitro. (A) GST-DLC-1 point mutants (Coomassie) were assayed for binding to full-length His₆-METT-10 proteins. WT, wild type. (B) METT-10 can form multimers. Western blot (WB) analysis of bacterially expressed His₆-METT-10 fusion proteins reveals bands that run at the size of monomers (*) and dimers ($). The lowest band in lane 1 likely represents a METT-10 degradation product. (C) A dimeric band (feathered arrow) is observed when full-length His₆-METT-10 is pulled down with GST-DLC-1 and full-length GST-METT-10. (D) Fine mapping of the DLC-1 binding domain of METT-10 using GST-pulldown analysis analogous to that shown in panel C. The identity of individually mutated METT-10 residues are labeled on the left.