| Products/Services Used | Details | Operation |
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| Bacterial Expression System | The genes for the E. Coli BirA biotin ligase and Avi–BMI1106–326 were ordered from Genscript and Life Technologies, respectively, and subcloned into the pCMV vector...For crystallization, BMI1 was incubated with two-fold excess of a synthetic PHC233–56 peptide (Genscript) and applied to a Superdex S75 gel filtration column (GE Healthcare) pre-equilibrated with storage buffer...Peptide PHC233–56 (Genscript) was directly dissolved in ITC buffer...Dissociation constants for binding of PHC2 to BMI1 UBL domain variants and MEL18 UBL were determined by fluorescence polarization. Fluorescein-labelled PHC232–61 (Genscript) at 20 nM was titrated | Get A Quote |
BMI1 is a core component of the polycomb repressive complex 1 (PRC1) and emerging data support a role of BMI1 in cancer. The central domain of BMI1 is involved in protein-protein interactions and is essential for its oncogenic activity. Here， we present the structure of BMI1 bound to the polyhomeotic protein PHC2 illustrating that the central domain of BMI1 adopts an ubiquitin-like (UBL) fold and binds PHC2 in a β-hairpin conformation. Unexpectedly， we find that the UBL domain is involved in homo-oligomerization of BMI1. We demonstrate that both the interaction of BMI1 with polyhomeotic proteins and homo-oligomerization via UBL domain are necessary for H2A ubiquitination activity of PRC1 and for clonogenic... More
