Jef Wagner

RNA topology remolds electrostatic stabilization of viruses

Simple RNA viruses efficiently encapsulate their genome into a nano-sized protein shell: the capsid. Spontaneous coassembly of the genome and the capsid proteins is driven predominantly by electrostatic interactions between the negatively charged RNA and the positively charged inner capsid wall. Using field theoretic formulation we show that the inherently branched RNA secondary structure allows viruses to maximize the amount of encapsulated genome and make assembly more efficient, allowing viral RNAs to out-compete cellular RNAs during replication in infected host cells.

Demonstration of angle-dependent Casimir force between corrugations.

The normal Casimir force between a sinusoidally corrugated gold coated plate and a sphere was measured at various angles between the corrugations using an atomic force microscope. A strong dependence on the orientation angle of the corrugation is found. The measured forces were found to deviate from the proximity force approximation and are in agreement with the theory based on the gradient expansion including correlation effects of geometry and material properties. We analyze the role of temperature. The obtained results open new opportunities for control of the Casimir effect in micromechanical systems.

The Robust Assembly of Small Symmetric Nanoshells.

Highly symmetric nanoshells are found in many biological systems, such as clathrin cages and viral shells. Many studies have shown that symmetric shells appear in nature as a result of the free-energy minimization of a generic interaction between their constituent subunits. We examine the physical basis for the formation of symmetric shells, and by using a minimal model, demonstrate that these structures can readily grow from the irreversible addition of identical subunits. Our model of nanoshell assembly shows that the spontaneous curvature regulates the size of the shell while the mechanical properties of the subunit determine the symmetry of the assembled structure. Understanding the minimum requirements for the formation of closed nanoshells is a necessary step toward engineering of nanocontainers, which will have far-reaching impact in both material science and medicine.

In vitro protease cleavage and computer simulations reveal the HIV-1 capsid maturation pathway

HIV-1 virions assemble as immature particles containing Gag polyproteins that are processed by the viral protease into individual components, resulting in the formation of mature infectious particles. There are two competing models for the process of forming the mature HIV-1 core: the disassembly and de novo reassembly model and the non-diffusional displacive model. To study the maturation pathway, we simulate HIV-1 maturation in vitro by digesting immature particles and assembled virus-like particles with recombinant HIV-1 protease and monitor the process with biochemical assays and cryoEM structural analysis in parallel. Processing of Gag in vitro is accurate and efficient and results in both soluble capsid protein and conical or tubular capsid assemblies, seemingly converted from immature Gag particles. Computer simulations further reveal probable assembly pathways of HIV-1 capsid formation. Combining the experimental data and computer simulations, our results suggest a sequential combination of both displacive and disassembly/reassembly processes for HIV-1 maturation.

Impact of a nonuniform charge distribution on virus assembly

Many spherical viruses encapsulate their genomes in protein shells with icosahedral symmetry. This process is spontaneous and driven by electrostatic interactions between positive domains on the virus coat proteins and the negative genomes. We model the effect of the nonuniform icosahedral charge distribution from the protein shell instead using a mean-field theory. We find that this nonuniform charge distribution strongly affects the optimal genome length and that it can explain the experimentally observed phenomenon of overcharging of virus and viruslike particles.

Effects of RNA branching on the electrostatic stabilization of viruses

Many single-stranded (ss) ribonucleic acid (RNA) viruses self-assemble from capsid protein subunits and the nucleic acid to form an infectious virion. It is believed that the electrostatic interactions between the negatively charged RNA and the positively charged viral capsid proteins drive the encapsidation, although there is growing evidence that the sequence of the viral RNA also plays a role in packaging. In particular, the sequence will determine the possible secondary structures that the ssRNA will take in solution. In this work, we use a mean-field theory to investigate how the secondary structure of the RNA combined with electrostatic interactions affects the efficiency of assembly and stability of the assembled virions. We show that the secondary structure of RNA may result in negative osmotic pressures while a linear polymer causes positive osmotic pressures for the same conditions. This may suggest that the branched structure makes the RNA more effectively packaged and the virion more stable.

Scattering approach for fluctuation-induced interactions at fluid interfaces.

We develop the scattering formalism to calculate the interaction between colloidal particles trapped at a fluid interface. Since, in addition to the interface, the colloids may also fluctuate in this system, we implement the fluctuation of the boundaries into the scattering formalism and investigate how the interaction between colloids is modified by their fluctuations. This general method can be applied to any number of colloids with various geometries at an interface. We apply the formalism derived in this work to a system of spherical colloids at the interface between two fluid phases. For two spherical colloids, this method very effectively reproduces the previous known results. For three particles we find analytical expressions for the large separation asymptotic energies and numerically calculate the Casimir interaction at all separations. Our results show an interesting three-body effect for fixed and fluctuating colloids. While the three-body effect strengthens the attractive interaction between fluctuating colloids, it diminishes the attractive force between colloids fixed at an interface.

Adsorption of annealed branched polymers on curved surfaces.

The behavior of annealed branched polymers near adsorbing surfaces plays a fundamental role in many biological and industrial processes. Most importantly single stranded RNA in solution tends to fold up and self-bind to form a highly branched structure. Using a mean field theory, we both perturbatively and numerically examine the adsorption of branched polymers on surfaces of several different geometries in a good solvent. Independent of the geometry of the wall, we observe that as branching density increases, surface tension decreases. However, we find a coupling between the branching density and curvature in that a further lowering of surface tension occurs when the wall curves towards the polymer, but the amount of lowering of surface tension decreases when the wall curves away from the polymer. We find that for branched polymers confined into spherical cavities, most of branch-points are located in the vicinity of the interior wall and the surface tension is minimized for a critical cavity radius. For branch polymers next to sinusoidal surfaces, we find that branch-points accumulate at the valleys while end-points on the peaks.

Three-body fluctuation-induced interaction at fluid interfaces: a strong deviation from the pairwise summation.

We present a new method based on the scattering technique to investigate fluctuation-induced forces at a fluid interface. The scattering approach, well suited to the study of many-body systems of arbitrary geometries, is augmented to include boundary fluctuations. Using this method, we study the deviation of the total fluctuation-induced interaction from the sum of pairwise energies for three colloidal particles. We consider both frozen and fluctuating colloids and obtain a very good agreement between analytical and numerical results. We find a marked difference in the three-body fluctuation-induced free energy between the frozen and fluctuating colloids, both in sign and relative size.

Role of genome in the formation of conical retroviral shells

Human immunodeficiency virus (HIV) capsid proteins spontaneously assemble around the genome into a protective protein shell called the capsid, which can take on a variety of shapes broadly classified as conical, cylindrical, and irregular. The majority of capsids seen in in vivo studies are conical in shape, while in vitro experiments have shown a preference for cylindrical capsids. The factors involved in the selection of the unique shape of HIV capsids are not well understood, and in particular the impact of RNA on the formation of the capsid is not known. In this work, we study the role of the genome and its interaction with the capsid protein by modeling the genomic RNA through a mean-field theory. Our results show that the confinement free energy for a homopolymeric model genome confined in a conical capsid is lower than that in a cylindrical capsid, at least when the genome does not interact with the capsid, which seems to be the case in in vivo experiments. Conversely, the confinement free energy for the cylinder is lower than that for a conical capsid if the genome is attracted to the capsid proteins as the in vitro experiments. Understanding the factors that contribute to the formation of conical capsids may shed light on the infectivity of HIV particles.