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Geometry Matters for Cargos Navigating 3D Microtubule Intersections

Creative Commons 'BY' version 4.0 license

Eukaryotic cells transport cargos along microtubules to control their distribution within the cell and deliver them to distant locations. While we understand how molecular motors can transport cargos along individual microtubules, the cell's microtubules are usually arranged in a complex 3D network. While traversing this network, cargos need to navigate intersections where microtubules cross at a wide variety of separation distances and angles. To gain insight into how cargos navigate these intersections, we have used a recently established 3D construction technique based on holographic optical trapping to build single 3D microtubule intersections in vitro with relevant nanoscale precision. We then used these fully suspended microtubule structures to perform motility assays on kinesin-1 coated cargos. We find that some intersection geometries influence cargos to pass along their current microtubule, while other geometries influence them to switch to the intersecting one. To understand how, we use a 3D Brownian dynamics simulation of cargo transport to investigate the mechanisms which give rise to the observed switching probabilities. Using these stochastic simulations, we find that switching probability is often determined by a competition between a stronger motor team on the primary microtubule and the intersecting microtubule sterically hindering that team's progress. This understanding of the basic mechanisms of switching at single intersections in 3D helps lay a foundation for understanding how the cell may regulate switching to control how cargos navigate the MT network and ultimately their spatial organization.

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