Mapping of the interacting domains of CLP36 and α-actinin-1 by use of the yeast 2-hybrid system.

(A) CLP36 and α-actinin-1 and their respective domains were constructed and cloned in “bait” and “prey” plasmids. (B) CLP36 (1-256) and CLP36 (81-256) interact strongly with a construct consisting of spectrinlike repeats 2-3 of α-actinin-1. β-Galactosidase activity of each transformant was visualized. The + indicates slow appearance (within 3-4 hours) of light blue colonies, +++ indicates rapid appearance (within 30-60 minutes) of dark blue colonies in the X-Gal assay on Trp⁻Leu⁻drop-out medium. The − indicates colonies that are white in the X-Gal screening assay; nt, interaction not tested.