Synthetic Biomolecular Condensates as Tunable Microtubule Assembly Hubs

Abstract Phase separation of proteins and nucleic acids (NAs) into nano-to-microscale condensates can regulate biochemical processes, including assembly and organization of cytoskeletal networks such as actin and microtubules. This study examines the functional role of condensate material properties in microtubule assembly. Learning from the sequence grammar of naturally occurring intrinsically disordered regions in microtubule-associated proteins, two-component peptide-NA condensates with programmable material properties were designed. These synthetic condensates catalyze tubulin polymerization into microtubule filaments with tunable outcomes. Tubulin preferentially partitions to the condensate interface and nucleates microtubule assembly. Enhanced tubulin self-assembly produces long filaments that exhibit branching and bundling. Using a minimal stochastic chemo-mechanical model, we show that sequence-encoded condensate viscoelasticity is a tunable element that controls filament morphologies and identifies interfacial rheology as the key regulator of filament growth. Fluorescence recovery after photobleaching experiments support this model, revealing a direct correlation between interfacial tubulin mobility and condensate-directed microtubule assembly. Distinct regimes emerge due to competition between bulk adsorption and lateral diffusion of tubulin at the condensate interface, which determines whether filament tips grow or stall. Since dynamic microtubule assembly and restructuring are essential for various cellular functions, this work highlights a critical role of condensate interfacial rheology in cytoskeletal organization. Competing Interest Statement P.R.B. is a member of the Biophysics Reviews (AIP Publishing) editorial board. This affiliation did not influence the work reported here. All other authors have no conflicts to report. Footnotes ↵# Lead contact: P.R.B. (prbanerj{at}buffalo.edu)