Mechanism of nuclear movements in a multinucleated cell

Supplemental Materials

This article contains the following supporting material:

  • Supplemental Materials
  • Supplemental Materials
  • Video 1 - VIDEO 1: Example of live-hyphae expressing GFP-tagged histone H4. DIC in gray and GFP signal in green.
  • Video 2 - VIDEO 2: A “wild-type” simulation with 3 cMTs per nucleus. These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 3 - VIDEO 3: A “wild-type” simulation with 6 cMTs per nucleus. These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 4 - VIDEO 4. A “wild-type” simulation with 3 cMTs per nucleus. The same simulation as shown on Video 1 is displayed here, using a different style. The organelles are not displayed, and the nuclei are semi-transparent such that the SPB (orange) is always visible. The cortical anchors are drawn in dark gray when they are unbound/inactive, and green when they are loaded with bound/active dynein. The re-orientation of the SPB upon pulling is clearly visible, for example for the right-most nucleus during the interval 1400?1700s. These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6BC;m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 5 - VIDEO 5. A simulation with bik1Δ?like MT dynamics. Everything is like the “wild-type” with 3 cMT/nucleus, except that MT growth and catastrophe rates were altered to represent the MT dynamics in bik1Δ mutants (see Figure 4A-B). These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 6 - VIDEO 6. A simulation with kip3Δ?like MT dynamics. Everything is like the “wild-type” with 3 cMT/nucleus, except that MT growth and catastrophe rates were altered to represent the MT dynamics in kip3Δ mutants (see Figure 4A-B). These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 7 - VIDEO 7. A simulation with S. cerevisiae?like MT dynamics. Everything is like the “wild-type” with 3 cMT/nucleus, except that MT growth and shrinkage as well as catastrophe rates were altered to represent the MT dynamics measured in S. cerevisiae (see Figure 4A-B). These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 8 - VIDEO 8. A simulation with a reduced number of cortical anchors. Everything is like the “wild-type” with 3 cMT/nucleus, except that the number of anchors on the cortex has been reduced to 100 instead of 200. These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 9 - VIDEO 9. A simulation with an even more reduced number of cortical anchors. Everything is like the “wild-type” with 3 cMT/nucleus, except that the staff number of cortical anchors has been reduced to 20 instead of 200. These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 10 - VIDEO 10. A “wild-type” simulation with 3 cMTs per nucleus and no cytoplasmic flow (speed = 0 μm/s.) These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 11 - VIDEO 11. A “wild-type” simulation with 6 cMTs per nucleus and no cytoplasmic flow (speed = 0 μm/s). These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 12 - VIDEO 12. A “wild-type” simulation with 3 cMTs per nucleus at a high cytoplasmic flow (speed = 0.02 μm/s). These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 13 - VIDEO 13. A “wild-type” simulation with 6 cMTs per nucleus at a high cytoplasmic flow (speed = 0.02 μm/s). These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 14 - VIDEO 14. An “empty” simulation without organelles. The simulation does not contain the extra spheres that are used to represent mitochondria and large spherical organelles. These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.
  • Video 15 - VIDEO 15. A simulation with “high” crowding. The simulation contains 550 spheres of 0.4 μm in diameter (orange, simulating mitochondria) and 50 beads of 1.4 ?m in diameter (blue, simulating large spherical organelles in old hyphae). These 14 movies represent various conditions, but the graphical representation is always identical, unless otherwise stated. The hypha (black) contains 5 nuclei (purple). One SPB (orange) on each nucleus may nucleate either 3 or 6 cMTs (white lines) that are elongating or shortening at the (distal) plusend. MTs may be pulled by dynein motors that load on fixed anchored at the cortex. The anchor points are displayed gray, and then green if a dynein is engaged. Forces produced by dynein motors and transmitted along the MTs can move the nucleus. The simulation uses periodic boundary conditions, and objects leaving on the right side reappear and the left side, and vice-versa. The cytoplasm is also filled with various organelles. Unless otherwise noted, the simulation contains 550 spheres of diameter 0.4BC;m (mitochondria, orange) and 50 spheres of diameter 0.6?m (blue, large spherical organelles) and a uniform cytoplasmic flow is moving all objects from left to right at a speed of 0.009 BC;m/s.