Interactions and functional roles of transcription factors involved in angiogenesis.

摘要

Transcription factors, FOXC2 (Forkhead C2) and ETV2 (Ets Variant 2), together play important roles in angiogenesis, and high expression of FOXC2 is reported in cancer stem cells. Expression of FOXC2 and ETV2 is necessary and sufficient for vascular development. Therefore, understanding the binding and interaction of the two proteins and their target DNA will facilitate development of therapeutic approaches. In this study, the DNA binding domains of FOXC2 and ETV2 proteins and the full length FOXC2 protein were expressed in a bacterial system and purified. High yield of the proteins was achieved through our established protocols. The complexes of the purified proteins and the target DNA were formed and verified using electrophoretic mobility shift assay. Fluorescent anisotropy experiments were performed to determine binding parameters of the full length FOXC2 with the DNA. Crystallization trials of the full length FOXC2 protein and DNA complex yielded initial crystallization conditions. To understand interactions among the proteins and DNA, a molecular model of FOXC2 and ETV2 was built using the FoxO1:Ets1:DNA complex structure. The reported studies will help to understand the interactions between FOXC2 and ETV and provide the foundation for developing inhibitors against these proteins to prevent angiogenesis in cancer cells.