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. 2004 Feb 12;23(4):719–727. doi: 10.1038/sj.emboj.7600111

Figure 4.

Figure 4

Labelling of the N- and C-terminal ends of TAF5. (A) Schematic representation of the structural features of TAF5 showing the WD-40 repeats and the 30 kDa tag for tandem affinity purification (TAP) that was introduced at the C-terminus of TAF5. (B) Average image of TFIID molecules in which TAF5 was fused to the TAP tag (left panel) and statistically significant density difference map with wild-type TFIID molecules (right panel). The difference map shows two additional densities located in lobes A and B. (C) Average images of TFIID containing TAP-tagged TAF5 subunits. The TAP tag was localized with an antibody to reveal the protein A moiety (left panels) and difference maps with nonlabelled TFIID molecules (right panels). Two distinct labelled particles were obtained that were labelled either in lobe B (upper panel) or in lobe A (lower panel), thus confirming the position of the TAP tag placed at the C-terminus of TAF5. (D) Average images of yTFIID molecules specifically labelled with a pAb raised against the first 18 residues of TAF5 (left panel) and corresponding difference map (right panel).