Supplemental Movies from my PhD thesis entitled "Organization and Dynamics of Treadmilling Filaments in Cytoskeletal Networks of FtsZ and its Crosslinkers"
Movie Captions
Movie 1 | FtsZ alone self-organization assayTime-lapse movie (raw data) of our self-organization assay showing FtsZ (gray) pattern emerging from its interaction with FtsA (unlabeled) alone. The architecture of these FtsZ/FstA co-filaments is typically characterized by dynamic networks of curved filament bundles, forming streams and vortexes. All the components were added to the reaction buffer (50mM Tris-HCl pH=7.5, 150mM KCl, 5mM MgCl2) prior to the addition of 4mM of GTP, which triggers FtsZ polymerization. Images were acquired using a TIRF microscope setup (typically one image every 2 seconds).
Movie 2 | FtsZ+ZapA 6uM self-organization assayTime-lapse movie (raw data) of our self-organization assay showing FtsZ pattern emerging from its interaction with FtsA and 6uM ZapA. FtsZ network organized into a network of thicker and longer filament bundles seemingly more alignedand straightened when compared with FtsZ alone. See Movie 1 caption for more details.
Movie 3 | Other ZapA concentrations self-organization assayTime-lapse movie (raw data) of our self-organization assay showing FtsZ pattern emerging from its interaction with FtsA in the presence of 0:5uM (left), 1:5uM (center) or 3:0uM (right) ZapA. Increasing concentrations of ZapA progressively increased the thickness of the FtsZ filament bundles and decreased its curvature. See Movie 1 caption for details.
Movie 4 | ZapA Mutants self-organization assayTime-lapse movie (raw data) of our self-organization assay showing FtsZ pattern emerging from its interaction with FtsA in the presence of 6uM ZapA R46A (left) and 6uM ZapA I83E (right). Mutants had no significant effect on the architecture and dynamics of FtsZ filament bundles. See Movie 1 caption for details.
Movie 5 | ZapB effect on ZapA activityTime-lapse movies (raw data) showing the inhibitory effect of ZapB on ZapA. Top row shows the effect of 1uM, 2uM and 6uM ZapB on 3uM ZapA (from left to right). Bottom row shows the effect of 3uM, 6uM and 12uM ZapB on 6uM ZapA (from left to right). ZapB prevented ZapA effect when [ZapB] >> [ZapA].
Movie 6 | FtsZ+ZapC 1uM self-organization assayTime-lapse movie (raw data) of our self-organization assay showing FtsZ pattern emerging from its interaction with FtsA and 1uM ZapC. FtsZ network organized into a carpet-li...
Movie Captions
Movie 1 | FtsZ alone self-organization assayTime-lapse movie (raw data) of our self-organization assay showing FtsZ (gray) pattern emerging from its interaction with FtsA (unlabeled) alone. The architecture of these FtsZ/FstA co-filaments is typically characterized by dynamic networks of curved filament bundles, forming streams and vortexes. All the components were added to the reaction buffer (50mM Tris-HCl pH=7.5, 150mM KCl, 5mM MgCl2) prior to the addition of 4mM of GTP, which triggers FtsZ polymerization. Images were acquired using a TIRF microscope setup (typically one image every 2 seconds).
Movie 2 | FtsZ+ZapA 6uM self-organization assayTime-lapse movie (raw data) of our self-organization assay showing FtsZ pattern emerging from its interaction with FtsA and 6uM ZapA. FtsZ network organized into a network of thicker and longer filament bundles seemingly more alignedand straightened when compared with FtsZ alone. See Movie 1 caption for more details.
Movie 3 | Other ZapA concentrations self-organization assayTime-lapse movie (raw data) of our self-organization assay showing FtsZ pattern emerging from its interaction with FtsA in the presence of 0:5uM (left), 1:5uM (center) or 3:0uM (right) ZapA. Increasing concentrations of ZapA progressively increased the thickness of the FtsZ filament bundles and decreased its curvature. See Movie 1 caption for details.
Movie 4 | ZapA Mutants self-organization assayTime-lapse movie (raw data) of our self-organization assay showing FtsZ pattern emerging from its interaction with FtsA in the presence of 6uM ZapA R46A (left) and 6uM ZapA I83E (right). Mutants had no significant effect on the architecture and dynamics of FtsZ filament bundles. See Movie 1 caption for details.
Movie 5 | ZapB effect on ZapA activityTime-lapse movies (raw data) showing the inhibitory effect of ZapB on ZapA. Top row shows the effect of 1uM, 2uM and 6uM ZapB on 3uM ZapA (from left to right). Bottom row shows the effect of 3uM, 6uM and 12uM ZapB on 6uM ZapA (from left to right). ZapB prevented ZapA effect when [ZapB] >> [ZapA].
Movie 6 | FtsZ+ZapC 1uM self-organization assayTime-lapse movie (raw data) of our self-organization assay showing FtsZ pattern emerging from its interaction with FtsA and 1uM ZapC. FtsZ network organized into a carpet-li...