Search for: All records

Crosslinked cytoskeletal filament networks provide cells with a mechanism to regulate cellular mechanics and force transmission. An example in the microtubule cytoskeleton is mitotic spindle elongation. The three-dimensional geometry of these networks, including the overlap length and lateral microtubule spacing, likely controls how forces can be regulated, but how these parameters evolve during filament sliding is unknown. Recent evidence suggests that the crosslinker PRC1 can resist microtubule sliding by two distinct modes: a braking mode and a less resistive coasting mode. To explore how molecular-scale mechanisms influence network geometry in this system, we developed a computational model of sliding microtubule pairs crosslinked by PRC1 that reproduces the experimentally observed braking and coasting modes. Surprisingly, we found that the braking mode was associated with a substantially smaller lateral separation between the crosslinked microtubules than the coasting mode. This closer separation aligns the PRC1-mediated forces against sliding, increasing the resistive PRC1 force and dramatically reducing sliding speed. The model also finds an emergent similar average sliding speed due to PRC1 resistance, because higher initial sliding speed favors the transition to braking. Together, our results highlight the importance of the three-dimensional geometric relationships between crosslinkers and microtubules. 

Kinesin-5 motors play an essential role during mitotic spindle assembly in many organisms: they crosslink antiparallel spindle microtubules, step toward plus ends, and slide the microtubules apart. This activity separates the spindle poles and chromosomes. Kinesin-5s are not only plus-end-directed, but can walk or be carried toward MT minus ends where they show enhanced localization. The kinesin-5 C-terminal tail interacts with and regulates the motor, affecting structure, motility, and sliding force of purified kinesin-535–37 along with motility and spindle assembly in cells. The tail contains phosphorylation sites, particularly in the conserved BimC box. Nine mitotic phosphorylation sites were identified in the kinesin-5 motor of the fission yeast Schizosaccharomyces pombe, suggesting that multi-site phosphorylation may regulate kinesin-5s. Here, we show that mutating all nine sites to either alanine or glutamate causes temperature-sensitive lethality due to a failure of bipolar spindle assembly. We characterize kinesin-5 localization and sliding force in the spindle, based on Cut7-dependent microtubule minus-end protrusions in cells lacking kinesin-14 motors. Imaging and computational modeling show that Cut7p simultaneously moves toward minus ends of protrusion MTs and plus ends of spindle midzone MTs. Phosphorylation mutants show dramatic decreases in protrusions and sliding force. Comparison to a model of force to create protrusions suggests that tail truncation and phosphorylation mutants decrease Cut7p sliding force similarly to tail-truncated human Eg5. Our results show that C-terminal tail phosphorylation is required for kinesin-5/Cut7 sliding force and bipolar spindle assembly in fission yeast. 

Abstract Flare frequency distributions represent a key approach to addressing one of the largest problems in solar and stellar physics: determining the mechanism that counterintuitively heats coronae to temperatures that are orders of magnitude hotter than the corresponding photospheres. It is widely accepted that the magnetic field is responsible for the heating, but there are two competing mechanisms that could explain it: nanoflares or Alfvén waves. To date, neither can be directly observed. Nanoflares are, by definition, extremely small, but their aggregate energy release could represent a substantial heating mechanism, presuming they are sufficiently abundant. One way to test this presumption is via the flare frequency distribution, which describes how often flares of various energies occur. If the slope of the power law fitting the flare frequency distribution is above a critical threshold,α= 2 as established in prior literature, then there should be a sufficient abundance of nanoflares to explain coronal heating. We performed >600 case studies of solar flares, made possible by an unprecedented number of data analysts via three semesters of an undergraduate physics laboratory course. This allowed us to include two crucial, but nontrivial, analysis methods: preflare baseline subtraction and computation of the flare energy, which requires determining flare start and stop times. We aggregated the results of these analyses into a statistical study to determine thatα= 1.63 ± 0.03. This is below the critical threshold, suggesting that Alfvén waves are an important driver of coronal heating.