FIG. 4.

Summary of pendulin deletions and corresponding LEF-1 binding activities. (A) The amino acid endpoints of the N- and C-terminal deletions are given for each construct and are shown relative to arm repeat borders that match those of Yano et al. (59). A summary of the activity of each deletion is given on the right for the in vitro binding assay and for the filter assay in the yeast two-hybrid system. n.t., not tested. (B) Filter assay for β-galactosidase activity in the yeast two-hybrid system. Each pendulin deletion (depicted in panel A) was cloned into a pACTI or pACTII yeast two-hybrid vector (gift of S. Elledge) in frame with sequences encoding the Gal4 transcription activation domain. The LEF-1 HMG DNA binding domain was inserted into a pAS vector to be expressed as a fusion protein with the Gal4 DNA binding domain. Both yeast two hybrid expression plasmids were cotransformed into strain Y190 and selected on Trp- and Leu-deficient selective medium for 4 days. An increase in the levels of β-galactosidase activity is an indication of the in vivo interaction between the two fusion proteins.