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. 2009 Nov;151(3):1498–1512. doi: 10.1104/pp.109.141705

Figure 6.

Figure 6.

Interactions of ZmSMU2 and AtSMU2 with other proteins. A, Y2H bait constructs were tested for self-activation and the corresponding β-GAL assays. A construct containing the full-length ZmSMU2 protein (pBD-GAL4-ZmSMU2; top) exhibited strong self-activation of the β-GAL reporter gene when introduced into Y190 cells. N-terminal deletions of ZmSMU2 (bait constructs I–IV) also showed self-activation, albeit at lower levels than for the full-length protein. Only a C-terminal deletion (bait construct V) and a negative control (pBD-GAL4; bottom) did not activate the reporter gene. Black bars represent the ZmSMU2 polypeptides. Numbers above the bars indicate amino acid residues corresponding to the full-length ZmSMU2 polypeptide. B, Confirmation of Y2H screen by in vitro pull-down assay using GST or GST fusion protein as a bait. GST (lane 2), GST-AtSMU2 (lane 3), or GST-ZmSMU2 (lane 4) bound to glutathione-agarose beads was used to pull down 6XHis-histone H4, 6XHis-AtSMU1, or 6XHis-AtMEC8. Lane 1, Ten percent input; lanes 2, 3, and 4, 30% pull-down by GST, GST-AtSMU2, and GST-ZmSMU2, respectively.