Figure 2.

BRD3 binds to a short motif in the N-terminal disordered region of CHD4. A, human CHD4 constructs used in this study. Domains with known structures are indicated. The presence or absence of an interaction with BRD3 is indicated by + or −, respectively. CD = chromodomain. B, BRD3 coimmunoprecipitates with the N-terminal third of CHD4. BRD3-HA (full-length, L or BD) and CHD4-FLAG (full-length, NT, M, or CT) constructs were coexpressed in HEK293 cells and applied to anti-FLAG-agarose beads. Western blottings using anti-HA or anti-FLAG antibodies are shown. C, CHD4(265–310) (fragment Ne) binds BRD3. Bacterially expressed CHD4 fragments immobilized on GSH beads were used to pull down mammalian-expressed HA-BRD3-L. Each Western blotting was visualized using α-HA and α-GST antibodies. PD = pulldown; FT = flow-through; * = unidentified band.