PRC1 resists microtubule sliding in two distinct resistive modes due to variations in the separation between overlapping microtubules.

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, such as the degree of overlap length or lateral microtubule spacing, likely controls how forces can be regulated, but how these parameters evolve during filament sliding is unknown. Recent evidence suggests that PRC1, a non-motor crosslinking protein of the MAP65 family, can resist microtubule sliding by two distinct modes: a braking mode in which microtubule sliding is significantly impeded and a less resistive coasting mode. To explore how molecular-scale mechanisms influence three-dimensional 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, which likely extends to other cytoskeletal architectures such as cilia.