A Non-Symmetric Reconstruction Technique for Transcriptionally-Active Viral Assemblies

Abstract

The molecular mechanisms by which RNA viruses coordinate their transcriptional activities are not fully understood. For rotavirus, an important pediatric gastroenteric pathogen, transcription occurs within a double-layered particle that encloses the viral genome. To date, there remains very little structural information available for actively-transcribing rotavirus double-layered particles, which could provide new insights for antiviral development. To improve our vision of these viral assemblies, we developed a new combinatorial strategy that utilizes currently available highresolution image processing tools. First, we employed a 3D classification routine that allowed us to sort transcriptionally-active rotavirus assemblies on the basis of their internal density. Next, we implemented an additional 3D refinement procedure using the most active class of DLPs. For comparison, the refined structures were computed in parallel by (1) enforcing icosahedral symmetry, and by (2) using no symmetry operators. Comparing the resulting structures, we were able to visualize the continuum that exists between viral capsid proteins and the viral RNA for the first time.

RETRACTED: High‐Resolution Imaging of Human Cancer Proteins Using Microprocessor Materials

“…We have previously used these microchips to analyze other protein structures. [22,24,25] The frames of the microchips were 2 mm × 2 mm in the x-and y-dimension and 0.3 mm in the z-dimension. Arrays of microwells were etched into the imaging windows that can vary in dimensions from 500 μm × 200 μm to 500 μm × 100 μm.…”

“…Silicon nitride (SiN)-based microprocessor chips, or microchips, provide pristine, flat surfaces and reliable physical properties. [21][22][23][24][25][26] Their ability to perform under extreme temperature conditions also makes them an ideal substrate for bioelectronic applications. Apertures composed of Si x N x are commonly used in electron-based imaging applications and their role in high-resolution studies is rapidly emerging due to the popularity of micro-electromechanical systems (MEMS).…”