Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Spontaneous Confinement of mRNA molecules at Biomolecular Condensate Boundaries

Version 1 : Received: 18 August 2023 / Approved: 22 August 2023 / Online: 22 August 2023 (16:35:13 CEST)

A peer-reviewed article of this Preprint also exists.

Perelman, R.T.; Schmidt, A.; Khan, U.; Walter, N.G. Spontaneous Confinement of mRNA Molecules at Biomolecular Condensate Boundaries. Cells 2023, 12, 2250. Perelman, R.T.; Schmidt, A.; Khan, U.; Walter, N.G. Spontaneous Confinement of mRNA Molecules at Biomolecular Condensate Boundaries. Cells 2023, 12, 2250.

Abstract

Cellular biomolecular condensates termed ribonucleoprotein (RNP) granules often are enriched in messenger RNA (mRNA) molecules relative to the surrounding cytoplasm. Yet, the spatial localization and diffusion of mRNAs in close proximity to phase separated RNP granules is not well understood. In this study, we performed single molecule fluorescence imaging experiments of mRNAs in live cells in the presence of two types of RNP granules, stress granules (SG) and processing bodies (PB), which are distinct in their molecular composition and function. We developed a photobleaching- and noise-corrected colocalization imaging algorithm that was employed to determine the accurate positions of individual mRNAs relative to the granule’s boundaries. We found that mRNAs are often localized at granule boundaries, an observation consistent with recently published data. We suggest that mRNA molecules become spontaneously confined at the RNP granule boundary similar to the adsorption of polymer molecules at liquid-liquid interfaces, which is observed in various technological and biological processes. We also suggest that this confinement could be due to a combination of intermolecular interactions associated with, first, the screening of a portion of the RNP granule interface by the polymer and, second, electrostatic interactions due to a strong electric field induced by a Donnan potential generated across the thin interface.

Keywords

membraneless organelles; mRNA; HILO microscopy; liquid-liquid phase separation; biomolecular condensates; RNP granules; colocalization; intermolecular interactions; Donnan potential

Subject

Biology and Life Sciences, Biophysics

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