Journal
VIRUSES-BASEL
Volume 12, Issue 8, Pages -Publisher
MDPI
DOI: 10.3390/v12080846
Keywords
self-assembly; virus capsid; non-nucleic acid cargo
Categories
Funding
- Indiana Clinical and Translational Sciences Institute
- National Institutes of Health [UL1TR002529]
- National Center for Advancing Translational Sciences, a Clinical and Translational Sciences Award
- Graduate Training Program in Quantitative and Chemical Biology [T32-GM109825]
- Graduate Training Program in Quantitative and Chemical Biology (Indiana University)
- Milton Taylor Virology Fellowship
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The assembly of alphavirus nucleocapsid cores requires electrostatic interactions between the positively charged N-terminus of the capsid protein (CP) and the encapsidated polyanionic cargo. This system differs from many other viruses that can self-assemble particles in the absence of cargo, or form empty particles. We hypothesized that the introduction of a mutant, anionic CP could replace the need for charged cargo during assembly. In this work, we produced a CP mutant, Minus 38 (M38), where all N-terminal charged residues are negatively-charged. When wild-type (WT) and M38 CPs were mixed, they assembled into core-like particles (CLPs). These empty particles were of similar size and morphology to WT CLPs assembled with DNA cargo, but did not contain nucleic acid. When DNA cargo was added to the assembly mixture, the amount of M38 CP that was assembled into CLPs decreased, but was not fully excluded from the CLPs, suggesting that M38 competes with DNA to interact with WT CPs. The composition of CLPs can be tuned by altering the order of addition of M38 CP, WT CP, and DNA cargo. The ability to produce alphavirus CLPs that contain a range of amounts of encapsidated cargo, including none, introduces a new platform for packaging cargo for delivery or imaging purposes.
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