Fig 8. E. chaffeensis TRP47 binds a G+C-rich motif.
(A) TRP47 was predicted to bind multiple G+C-rich motifs. The motifs incorporated into probes P1, P2, P3, and P4, (two motifs per probe, underlined in the probe sequences in Table 1) are shown in WebLogo format. (B) The predicted motifs were tested by EMSA. Incubation of biotin-labeled probes P1, P2, P3, and P4 with full-length TRP47 resulted in a shifted band complex CTRP47 for P4 (lane 8 in left panel and lane 2 in right panel, marked by arrows). The relative density of the shifted band was diminished with addition of 100 and 250-fold molar excess of unlabeled competitor DNA (right panel, lane 3 and 4 respectively). This band was also reduced with addition of anti-TRP47 antibody, but accompanied by a supershifted band complex SCTRP47 (lane 5, marked by arrow). These results suggest that P4 contains a specific TRP47 DNA-binding motif.