Structural characterization of the interaction between TFIIIB components Bdp1 and Brf1

Transcription factor TFIIIB plays key roles in transcription by RNA polymerase III. Its three components (TBP, Brf1, and Bdp1) participate in crucial molecular events that include RNA polymerase recruitment, formation of the open initiation complex, and recycling of transcription. Although the details of the interaction among DNA, TBP, and Brf1 have been, in part, revealed through the crystal structure of their ternary complex, structural details of the Brf1-Bdp1 interaction are lacking. In this paper, nuclear magnetic resonance (NMR) is used to map the interaction interface between Bdp1 and Brf1 at single-amino acid resolution, using minimal functional segments of the two proteins. An NMR-derived structural model shows that the principal anchorage site of Brf1 is located on a convex surface of Bdp1 that encompasses helix 1 and helix 3 of its conserved SANT domain. The main Bdp1 anchorage site is provided by a small set of residues belonging to a Brf1 segment of residues 470-495.