Biology subjects

Liese, S.

Publications and source records attributed to Liese, S..

8 recordsLinked to original sources

bioRxiv2026

Thermodynamic Model and Experiments Indicate Kinetically Trapped States in Ionisable Lipid Nanoparticles

Lipid nanoparticles (LNPs) are the leading delivery vehicles for RNA-based therapies, with pH-dependent mesophase transitions of their ionizable lipids proposed to play a crucial role in LNP function and endosomal escape. However, a consistent quantitative framework linking molecular lipid properties to the thermodynamics of these transitions is still lacking. Here, we establish such a framework by combining X-ray scattering, fluorescence spectroscopy, all-atom molecular dynamics simulations, and continuum modeling. Using small-angle X-ray scattering (SAXS) and Laurdan fluorescence spectroscopy, we characterize the pH- and temperature-dependent phase behavior of both bulk lipid systems and LNPs containing the ionisable lipids MC3 or ALC-0315. Laurdan fluorescence measurements reveal the same pH- and temperature-dependent transitions in bulk and LNP systems, which closely follow the inverse micellar (LII) - inverse hexagonal (HII) transitions identified by SAXS, establishing Laurdan as a reporter of these mesophase transitions in LNPs. We combine experimental and simulation data to parameterize a continuum model of the LII - HII transition, providing a quantitative description of mesophase stability. The model reveals that geometric frustration arising from packing constraints is a key determinant of mesophase stability. By combining experimental and simulation data with theoretical analysis, we determine the relevant membrane parameters, including mean and Gaussian curvature bending rigidities, and identify the parameter ranges that govern phase stability. Together, these results establish a quantitative continuum theory for pH-dependent mesophase transitions of ionisable lipid and suggest metastable states in response to pH changes as an inherent feature of LNP core phase.

biophysics↗

bioRxiv2024

Cryo-electron tomography reveals coupled flavivirus replication, budding and maturation

Flaviviruses replicate their genomes in replication organelles (ROs) formed as bud-like invaginations on the endoplasmic reticulum (ER) membrane, which also functions as the site for virion assembly. While this localization is well established, it is not known to what extent viral membrane remodeling, genome replication, virion assembly, and maturation are coordinated. Here, we imaged tick-borne flavivirus replication in human cells using cryo-electron tomography. We find that the RO membrane bud is shaped by a combination of a curvature-establishing coat and the pressure from intraluminal template RNA. A protein complex at the RO base extends to an adjacent membrane, where immature virions bud. Naturally occurring furin site variants determine whether virions mature in the immediate vicinity of ROs. We further visualize replication in mouse brain tissue by cryo-electron tomography. Taken together, these findings reveal a close spatial coupling of flavivirus genome replication, budding, and maturation.

microbiology↗

bioRxiv2022

Architecture of the chikungunya virus replication organelle

Alphaviruses are mosquito-borne viruses that cause serious disease in humans and other mammals. Along with its mosquito vector, the alphavirus chikungunya virus (CHIKV) has spread explosively in the last 20 years, and there is no approved treatment for chikungunya fever. On the plasma membrane of the infected cell, CHIKV generates dedicated organelles for viral RNA replication, so-called spherules. Whereas structures exist for several viral proteins that make up the spherule, the architecture of the full organelle is unknown. Here, we use cryo-electron tomography to image CHIKV spherules in their cellular context. This reveals that the viral protein nsP1 serves as a base for the assembly of a larger protein complex at the neck of the membrane bud. Biochemical assays show that the viral helicase-protease nsP2, while having no membrane affinity on its own, is recruited to membranes by nsP1. The tomograms further reveal that full-sized spherules contain a single copy of the viral genome in double-stranded form. Finally, we present a mathematical model that explains the membrane remodeling of the spherule in terms of the pressure exerted on the membrane by the polymerizing RNA, which provides a good agreement with the experimental data. The energy released by RNA polymerization is found to be sufficient to remodel the membrane to the characteristic spherule shape.

biophysics↗

bioRxiv2020

Membrane Shape Remodeling by Protein Crowding

The steric repulsion between proteins on biological membranes is one of the most generic mechanisms that cause membrane shape changes. We present a minimal model where a spontaneous curvature is induced by steric repulsion between membrane-associated proteins. Our results show that the interplay between the induced spontaneous curvature and the membrane tension determine the energy minimizing shapes, which describe the wide range of experimentally observed membrane shapes, i.e. flat membranes, spherical vesicles, elongated tubular protrusions, and pearling structures.

biophysics↗

bioRxiv2020

Protein diffusion on membrane domes, tubes and pearling structures

Diffusion is a fundamental mechanism for protein distribution in cell membranes. These membranes often exhibit complex shapes, which range from shallow domes to elongated tubular or pearl-like structures. Shape complexity of the membrane influences the diffusive spreading of proteins and molecules. Despite the importance membrane geometry plays in these diffusive processes, it is challenging to establish the dependence between diffusion and membrane morphology. We solve the diffusion equation numerically on various curved shapes representative for experimentally observed membrane shapes. Our results show that membrane necks become diffusion barriers. We determine the diffusive half time, i.e., the time that is required to reduce the amount of proteins in the budded region by one half and find a quadratic relation between the diffusive half time and the averaged mean curvature of the membrane shape. Our findings thus help to estimate the characteristic diffusive time scale based on the simple measure for membrane morphology. Significance statementDiffusion is an integral process for distributing proteins throughout biological membranes. These membranes can have complex shapes and structures, often featuring elongated shapes such as tubes and like a necklace of pearls. The diffusion process on these shapes is significantly different from the well studied planar substrate. We use numerical simulations to understand how the characteristic diffusion time is a function of membrane shape, where we find the diffusion of proteins on strongly curved shapes is significantly slower than on planar membranes. Our results provide a simple relationship to estimate the characteristic diffusion time of proteins on membranes based on its mean and Gaussian curvature.

biophysics↗

bioRxiv2020

Rapid growth and fusion of protocells in surface-adhered membrane networks

Elevated temperatures might have promoted the nucleation, growth and replication of protocells on the early Earth. Recent reports have shown evidence that moderately high temperatures not only permit protocell assembly at the origin of life, but could have actively supported it. Here we show the fast nucleation and growth of vesicular compartments from autonomously formed lipid networks on solid surfaces, induced by a moderate increase in temperature. Branches of the networks, initially consisting of self-assembled interconnected nanotubes, rapidly swell into microcompartments which can spontaneously encapsulate RNA fragments. The increase in temperature further causes fusion of adjacent network-connected compartments, resulting in the redistribution of the RNA. The experimental observations and the mathematical model indicate that the presence of nanotubular interconnections between protocells facilitates the fusion process.

biophysics↗

bioRxiv2019

Protein Crowding mediates Passive Membrane Remodeling in ESCRT-induced Formation of Intraluminal Vesicles

As part of the lysosomal degradation pathway, the endosomal sorting complexes required for transport (ESCRT-0-III/VPS4) sequester receptors at the endosome and simultaneously deform the membrane to generate intraluminal vesicles (ILVs). Whereas ESCRT-III/VPS4 have an established function in ILV formation, the role of upstream ESCRTs (0-II) in membrane shape remodeling is not understood. Combining experimental measurements and electron microscopy analysis of ESCRT-III depleted cells with a mathematical model, we show that upstream ESCRT-induced alteration of the Gaussian bending rigidity and their crowding on the membrane induces membrane deformation and facilitates ILV formation: upstream ESCRT-driven budding does not require ATP consumption as only a small energy barrier needs to be overcome. Our model predicts that ESCRTs do not become part of the ILV, but localize with a high density at the membrane neck, where the steep decline in the Gaussian curvature likely triggers ESCRT-III/VPS4 assembly to enable neck constriction and scission.Significance Statement Intraluminal vesicle (ILV) formation plays a crucial role in the attenuation of growth factor receptor signaling, which is mediated by the endosomal sorting complex required for transport (ESCRT-0-III/VPS4). The general dogma has been that the upstream ESCRTs (0-II) sequester the receptors at the surface of endosomes and the downstream ESCRTs (III/VPS4) remodel the endosome membrane leading to the abscission and formation of receptor-containing ILVs. We now show that the upstream ESCRTs not only sequester cargo, but in addition play an essential role for the initiation of membrane shape remodeling in ILV budding. Through a combination of mathematical modeling and experimental measurements we show that upstream ESCRTs facilitate ILV budding by crowding with a high density in the membrane neck region.Competing Interest StatementThe authors have declared no competing interest.View Full Text

biophysics↗

bioRxiv2018

A nanotube-mediated path to protocell formation

Cellular compartments are membrane-enclosed, spatially distinct microenvironments which confine and protect biochemical reactions in the biological cell. On the early Earth, the autonomous formation of compartments is thought to have led to the encapsulation of nucleotides, thereby satisfying a starting condition for the emergence of life. Recently, surfaces have come into focus as potential platforms for the self-assembly of prebiotic compartments, as significantly enhanced vesicle formation was reported in the presence of solid interfaces. The detailed mechanism of such formation at the mesoscale is still under discussion. We report here on the spontaneous transformation of solid surface-adhered lipid deposits to unilamellar membrane compartments through a straightforward sequence of topological changes, proceeding via a network of interconnected lipid nanotubes. We show that this transformation is entirely driven by surface-free energy minimization and does not require hydrolysis of organic molecules, or external stimuli such as electrical currents or mechanical agitation. The vesicular structures take up and encapsulate their external environment during formation, and can subsequently separate and migrate upon exposure to hydrodynamic flow. This may link, for the first time, the self-directed transition from weakly organized bioamphiphile assemblies on solid surfaces to protocells with secluded internal contents.\n\nSignificanceThe nature of the physical and chemical mechanisms behind the formation, growth and division of the earliest protocells is among the key questions concerning the origin of life. Establishing a simple pathway for the assembly of protocell structures from the primordial soup is a particular challenge. Emerging evidence supporting the assumption that solid surfaces have a governing role in protocell formation has recently expanded the scope, and created new inspiration for investigation. By presenting a physical path from self-assembled amphiphile-based membranes on solid surfaces to spherical single-membrane compartments via a consistent sequence of transformations, solely driven by the materials properties of the interfaces, a direct link between the presence of functional biomolecules and the development of protocells can be established.

biophysics↗