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Fig. 6. Structural interaction between Topi and Comr proteins. (A) Mapping domains
of Comr responsible for the interaction with full-length Topi. Solid bars
represent protein fragments tested in the yeast two-hybrid system against
Topi. The region responsible for the interaction lies in the 200-300 amno acid
fragment (bracket). (B) Mapping domains of Topi responsible for the
interaction with full-length Comr. Zn-finger motifs within Topi are indicated
with boxes. Fragments containing either the first two, or the final five, Zn
fingers can bind Comr (brackets). (C) Co-immunoprecipitation of Comr and Topi
from tissue culture. Cells expressed HA-tagged Topi and FLAG-tagged Comr or
Smad1. Binding was assessed by IP-ing with anti-FLAG, and blotting with
anti-HA (first pair of lanes) or vice versa (second pair of lanes). Topi and
Comr co-immunoprecipitate in both cases (top). Total protein expression was
assessed by western blotting of the cell lysate (bottom).
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