Abstract
The functions performed by the concentric shells of multilayered dsRNA viruses require specific protein interactions that can be directly explored through their mechanical properties. We studied the stiffness, breaking force, critical strain and mechanical fatigue of individual Triple, Double and Single layered rotavirus (RV) particles. Our results, in combination with Finite Element simulations, demonstrate that the mechanics of the external layer provides the resistance needed to counteract the stringent conditions of extracellular media. Our experiments, in combination with electrostatic analyses, reveal a strong interaction between the two outer layers and how it is suppressed by the removal of calcium ions, a key step for transcription initiation. The intermediate layer presents weak hydrophobic interactions with the inner layer that allow the assembly and favor the conformational dynamics needed for transcription. Our work shows how the biophysical properties of the three shells are finely tuned to produce an infective RV virion.
Highlights
The advent of single-molecule techniques have opened the door to understand how the mechanics of biomolecular assemblies is essential for their function (Howard, 2001; Muller et al, 2002)
Previous studies have shown that RV triple-layered particle (TLP) can be converted to double-layered particle (DLP) by disassembling the outer VP4VP7 layer with chelating agents such as ethylenediaminetetraacetic acid (EDTA) (Estes et al, 1979)
DLP can be converted to single-layered particle (SLP) by chaotropic agents such as CaCl2 (Figure 1B) (Bican et al, 1982)
Summary
The advent of single-molecule techniques have opened the door to understand how the mechanics of biomolecular assemblies is essential for their function (Howard, 2001; Muller et al, 2002). In the case of viruses, the infectious particle must be robust enough to protect the viral genome outside the cell and competent to undergo the required structural changes once the host cell is recognized, overcome its barriers and carry out the events necessary for a productive viral replication cycle (Flint et al, 2004). Since there are no host cell enzymes that can recognize dsRNA as template for transcription, the viral particle must incorporate a transcription machinery able to synthesize the required mRNAs to initiate the viral replication cycle. Most dsRNA viruses exhibit a common solution to these problems, which consists of the assembly of a stable protein cage in the host cytoplasm that isolates the viral dsRNA molecules to prevent the cellular
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