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Competing Crosstalk between Cytoskeletal Filaments Dictates Structure and Superdiffusivity of Microtubules in Live Cells

Renita Saldanha1,2, Yiling Lan2,3, Heidi Hehnly2,3, Ryan J. McGorty4, Rae M. Robertson-Anderson4, and Alison Patteson1,2

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PRX Life 3, 033009 – Published 5 August, 2025

DOI: https://doi.org/10.1103/15vm-4d7b

Abstract

The cytoskeleton is an active and complex composite network of actin, microtubules, and intermediate filaments (IFs). Networks of these distinct biopolymers work in partnership to fulfill fundamental cellular processes, such as cell polarization, intracellular transport, and cell migration. While there is evidence that the different cytoskeletal filaments interact through biochemical signaling and physical interactions, many basic questions remain regarding how this crosstalk impacts the dynamics and organization of the cytoskeletal filaments. Here, we combine Fourier image analysis methods with live cell experiments to characterize the impacts of cytoskeletal crosstalk on the structure and dynamics of microtubules in mouse embryonic fibroblasts. Specifically, we investigate the impacts of depleting vimentin IFs (vim−/−) as well as inhibiting actomyosin activity or inducing actin depolymerization. Using spatial image autocorrelation analysis, we identify that depleting vimentin decreases the effective mesh size of the microtubule network regardless of the state of the actin, while inhibiting myosin activity or depolymerizing actin increases the microtubule mesh size in cells with and without vimentin. Moreover, using differential dynamic microscopy, we reveal that microtubules exhibit superdiffusive motion over a wide range of length- and timescales, with rates that are enhanced in all cells lacking vimentin and suppressed in all cells where actin is disrupted. Our results reveal competing and seemingly independent roles that vimentin and actin play in mediating the dynamics and structure of the network of microtubules in cells across decades of spatiotemporal scales. We argue that these effects arise from the scaffolding ability of the vimentin and active dynamics of the actin, which independently sculpt microtubule network dynamics and structure in cells.

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PRX Life 3, 033009– Published 5 August, 2025
  • Received 19 January 2025
  • Accepted 7 July 2025
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Published by the American Physical Society under the terms of the Creative Commons Attribution 4.0 International license. Further distribution of this work must maintain attribution to the author(s) and the published article's title, journal citation, and DOI.

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