Virus Assembly Pathways: Straying Away but Not Too Far

Abstract Non‐enveloped RNA viruses pervade all domains of life. In a cell, they co‐assemble from viral RNA and capsid proteins. Virus‐like particles can form in vitro where virtually any non‐cognate polyanionic cargo can be packaged. How only viral RNA gets selected for packaging in vivo, in presence of myriad other polyanionic species, has been a puzzle. Through a combination of charge detection mass spectrometry and cryo‐electron microscopy, it is determined that co‐assembling brome mosaic virus (BMV) coat proteins and nucleic acid oligomers results in capsid structures and stoichiometries that differ from the icosahedral virion. These previously unknown shell structures are strained and less stable than the native one. However, they contain large native structure fragments that can be recycled to form BMV virions, should a viral genome become available. The existence of such structures suggest the possibility of a previously unknown regulatory pathway for the packaging process inside cells.