DNA binding specificity of other DMRT proteins. (A) Summary of in vitro random oligonucleotide binding site selections for DMRT1, 2, 3, 4, 5, and 7. Sequence preference for DSX is from Yi et al., 1999 , reanalyzed using the Gibbs Motif Sampler program [35, 36]. Sequence preferences are displayed using the WebLogo program [44, 45] to indicate preferences at each position, with size of letter indicating information content of each nucleotide (bits). The selected sequences from which each consensus was derived are shown in Supplemental Figure 1. (B,C) Gel mobility shift analysis. (B) DMRT3 and DMRT5. (C) DMRT2, DMRT4, DMRT7. Labeled probes used are indicated below each panel, and proteins and antibodies present in binding reactions are indicated above. Mobility of dimer and monomer complexes with DNA are indicated for each protein ("M" or "D"); "NS" indicates non-specific complexes. Exposure time for panel C is 28-fold longer than for panel B. (D) DNA recognition by DMRT proteins in cultured cells. Transactivation by VP16 fusion proteins in HEK293 cells transfected with expression vectors encoding the indicated DMRT-VP16 fusions and with the pGL3-Promoter reporter plasmid containing a single DMRT consensus sequence upstream of the luciferase coding sequence. Values shown are average of two experiments, with error bar indicating standard error of the mean.
Murphy et al. BMC Molecular Biology 2007 8:58 doi:10.1186/1471-2199-8-58