Figure 1.

TAF_II55 binds TAF_II250. (A) The TAF_II250 fragment (amino acids 848-1279), spanning the AT and RAPiDs, was used as bait in a yeast two-hybrid screen to isolate TAF_II55 clones. The location of the bait fragment, relative to the full-length molecule, is shown. TAF_II250 AT domain (shaded box) is located approximately between amino acids 640 and 1093, which corresponds to the Drosophila AT domain (amino acids 612-1140) (18). (B) TAF_II55 clones isolated in yeast two-hybrid screens. The locations of the five isolated TAF_II55 clones are shown relative to the full-length human and mouse proteins. Although the clones differed in their 5′ termini, they shared a common 3′ end that extended to encode the carboxyl terminus of the TAF_II55 protein. The mouse TAF_II55 differs from the human in a deletion spanning amino acids 233–242. (C) TAF_II55 binds to the RAPiD of TAF_II250. Flag-tagged TAF_II55 was used to assess its ability to bind to either [³⁵S]methionine labeled, in vitro translated AT, or RAPiD domains, as shown in A (AT, lanes 2, 5, 8; RAPiD, lanes 1, 4, 7). In vitro translated SNAP23 protein was used as a control for nonspecific binding (lanes 3, 6, 9). The relative binding of the TAF_II250 fragments to TAF_II55 was quantitated and calculated relative to input, as shown beneath the lanes. Arrowheads mark the positions of input proteins.