Multiscale Growth Kinetics of Model Biomolecular Condensates Under Passive and Active Conditions

Living cells exhibit a complex organization comprising numerous compartments, among which are RNA- and protein-rich membraneless, liquid-like organelles known as biomolecular condensates. Energy-consuming processes regulate their formation and dissolution, with (de-)phosphorylation by specific enzymes being among the most commonly involved reactions. By employing a model system consisting of a phosphorylatable peptide and homopolymeric RNA, we elucidate how enzymatic activity modulates the growth kinetics and alters the local structure of biomolecular condensates. Under passive condition, time-resolved ultra-small-angle X-ray scattering with synchrotron source reveals a nucleation-driven coalescence mechanism maintained over four decades in time, similar to the coarsening of simple binary fluid mixtures. Coarse-grained molecular dynamics simulations show that peptide-decorated RNA chains assembled shortly after mixing constitute the relevant subunits. In contrast, actively-formed condensates initially display a local mass fractal structure, which gradually matures upon enzymatic activity before condensates undergo coalescence. Both types of condensate eventually reach a steady state but fluorescence recovery after photobleaching indicates a peptide diffusivity twice higher in actively-formed condensates consistent with their loosely-packed local structure. We expect multiscale, integrative approaches implemented with model systems to link effectively the functional properties of membraneless organelles to their formation and dissolution kinetics as regulated by cellular active processes.

Tamizhmalar Sundararajan, Matteo Boccalini, Roméo Suss, Sandrine Mariot, Emerson R. Da Silva, Fernando C. Giacomelli, Austin Hubley, Theyencheri Narayanan, Alessandro Barducci, Guillaume Tresset · 2025