FIG 3.

Y102E does not bind the Brd4 CTM but associates with an internal E2-binding region. (A) Domains of the Brd4 chromatin modulator protein (39). BD1, bromodomain 1; BID, basic interacting domain; ET, extraterminal domain. (B) HEK293TT cells were transfected with mRFP-GFP (−), the BPV-1 E2 WT and mutants (YF and YE), and FLAG-tagged full-length Brd4 (pVL-F:hBrd4) or pCI. Immunoprecipitates were collected with anti-FLAG mouse antibody M2 conjugated to agarose beads. Samples were probed by Western blotting for FLAG-Brd4 and BPV-1 E2 using M2 and B201, respectively. E2, full-length BPV-1 E2; *, nonspecific band. (C and D) GST-tagged CTM (C) and GST-BID (D). Complexes were collected with glutathione beads. Western blots were probed for GST with rabbit polyclonal antibody SD8 and for E2 with B201. *, nonspecific band.