Fig. 7

FOXM1 transcriptional activity requires direct chromatin interaction involving recruitment to non-consensus sequences. a qPCR analysis of the mRNA transcript levels in GFP-FOXM1 WT or mutant (H287A or R286A) cell lines treated ± doxycycline (1 μg/mL) for 24 h showing the relative change in the levels of FOXM1B, AURKB, CCNB1, CDC25B, CENPF, and PLK1. In each case the data are normalized to the minus doxycycline control. b Binding curves measured by fluorescence polarization analysis (assay details in the Materials and methods section), showing binding affinity of GST-FOXM1B DBD for 16-mer [FAM]dsDNA sequences present in FOXM1 binding peaks from the ChIP-seq dataset compared to the FKH consensus. The plot shows the fraction bound with increasing protein concentration. The table shows the K _d values ± SD determined for each sequence. c Illustration of alternative models proposed the recruitment of FOXM1 to chromatin. (1) Direct DNA binding of FOXM1 at promoter sites containing a FKH consensus motif and interaction with MuvB and B-Myb. (2) FOXM1 is recruited by MuvB complex and does not directly bind to the DNA. (3) FOXM1 binds directly at non-consensus sequences facilitated by interaction with MuvB and B-Myb. Arrow indicates transcription start site of target gene. Data representative of triplicate experiments ± SD. (*) P <0.05, (**) P <0.01, (***) P <0.001