Figure 5.

Dimerization of CMS is mediated through the COOH-terminal CC region. (A) Helical wheel representation of the putative leucine zipper in the CC region of CMS. (B) 293T cells were cotransfected with either empty vectors, GST vector, and flag-tagged CMS-TH (a), FLAG vector and GST-tagged CMS/P-CC (b), or CMS-TH and CMS/P-CC. Immunoprecipitates (IP) were analyzed by WB using anti-flag antibody (Upper). The expression levels were analyzed in total cell lysates by WB. (C) Mapping of the dimerization site in CMS. CMS target constructs for expression in yeast were introduced in vector pACT2 and transfected together with the bait CMS/P-CC cloned in pAS2–1. Transformants were grown on plates lacking leucine and tryptophane. Three single colonies were analyzed for interaction in a β-galactosidose filter assay (19). −, no interaction. +, interaction.