Download or read book Reovirus Core Proteins Play Roles in Disassembly and Early Replication written by Stephanie L. Gummersheimer and published by . This book was released on 2022 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Dissassembly of viral outer capsid is a critical step for successful infection of host cells. Mammalian orthoreovirus (reovirus) particles unable to complete disassembly steps upon entry have significantly reduced infectivity. Reovirus capsids contain two concentric protein shells that protect a segmented dsRNA genome. The outer capsid layer is composed of trimers of u1 capped with o3. The inner layer or core is composed of decamers of A1 stabilized by the clamp-like protein o2. After entry into host cells, o3 is proteolytically degraded and u1 is cleaved resulting in the formation of infectious subvirion particles or ISVPs. Further conformational changes in the capsid proteins result in the formation of a second disassembly intermediate, the ISVP. Previous work to characterize these disassembly intermediates has focused on changes and interactions between the outer capsid proteins. My work aims to examine the contributions of the core proteins to the disassembly process. To do this, we generated a series of reassortant viruses containing primarily gene segments from a type 3 reovirus except for the gene segments coding for core proteins A1 and o2 (L3 and S2 respectively) which came from a type 1 virus. To determine the residues important for interactions between the core and the outer capsid that could be contributing to this phenotype, we used two approaches. First, we screened for viruses that accumulated mutations that generate hyperstable ISVPs. Through this screen, we did not isolate residues in the core proteins important for these interactions but instead identified several amino acid residues in the u1 o region that had not been previously associated with ISVP stability. Our second approach focused on targeting specific amino acids residues in the core proteins that vary between type 3 and type 1 serotypes. We generated viruses with chimeric h1 proteins or with point mutations in o2. Via this method, we determined specific changes in the core proteins that reduced the stability of ISVPs. Additionally, using these specifically generated mutants, we identified regions h1 and o2 that affect events in viral replication distinct from entry. These data highlight the multiple roles for capsid proteins during infection.