A Non-Symmetric Reconstruction Technique for Transcriptionally-Active Viral Assemblies
Autor: | Andrew C. Demmert, Linda A. Melanson, Deborah F. Kelly, Amina Rahimi, A. Cameron Varano, Sarah M. McDonald |
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Rok vydání: | 2015 |
Předmět: |
Rotavirus
0303 health sciences Double-layered particles Cryo-electron microscopy Icosahedral symmetry viruses 030302 biochemistry & molecular biology Non symmetric Affinity capture RNA Computational biology Biology medicine.disease_cause Genome Virology Article 03 medical and health sciences Capsid Transcription (biology) medicine Transcription 030304 developmental biology |
Zdroj: | Journal of analytical & molecular techniques |
ISSN: | 2474-1914 |
Popis: | 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 high-resolution 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. |
Databáze: | OpenAIRE |
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