SlmA Antagonism of FtsZ Assembly Employs a Two-pronged Mechanism like MinCD
Figure 4
FtsZ-K190V and FtsZ-D86N bind SlmA.
A) SBS17-30mer bound SlmA co-sediments with stable FtsZ polymers formed with GMPCPP. Polymerization assays were performed as in Fig. 3A except that the protein concentration was 5 µM and GTP was replaced by GMPCPP. After the reactions were incubated at room temperature for 5 minutes, FtsZ polymers were sedimented by ultracentrifugation. Proteins in the supernatant and pellet fractions were separated by SDS-PAGE. B) Biolayer interferometry assay to assess FtsZ binding to SlmA bound to DNA. Streptavidin biosensor tips loaded with biotin conjugated SBS17-30mer and SlmA were incubated with FtsZ or the FtsZ mutants (4 µM) and the association monitored.