FIG. 6.

Hec1p interacts with Smc2p. (A) Domain mapping and temperature sensitivity of the interaction between Hec1p and Smc2p. The same constructs of Hec1p as used for Fig. 5 were used in yeast two-hybrid assays to test their binding ability to yeast Smc2p expressed as a Gal4 TAD fusion protein. Binding ability was tested at 25 or 37°C. Gal4-DBD, Gal4 DNA-binding domain; ND, not determined. (B) In vivo interaction between Hec1p and Smc2p. Equal amounts of cell lysates from the log-phase culture of strain Y153 alone (lanes 1 to 3) or Y153 expressing TAD-Smc2p (lanes 4 to 6) were immunoprecipitated (IP) by the antibodies indicated. The resultant immunocomplexes were separated by SDS-PAGE followed by immunoblotting. The upper panel was blotted by polyclonal anti-TAD antibodies, and the lower panel was stained by anti-scHec1p. Ig, immunoglobulin.