Subunits fold at position-dependent rates during maturation of a eukaryotic RNA virus.
Effective antiviral agents are difficult to develop because of the close relationship between the cell biology of the virus and host. However, viral capsid maturation, the in vivo process where the particle transitions from a noninfectious provirion to an infectious virion, is an ideal process to interrupt because the provirion is usually fragile and the conversion to the virion often involves large conformational changes and autocatalytic chemistry that can be hampered by small molecules. The Nudaurelia capensis omega virus (N omegaV) is one of the few eukaryotic viruses where this process can be investigated in vitro with a variety of biophysical methods, allowing fundamental chemical and structural principles of the maturation to be established. It has a T = 4 quasi-equivalent capsid with a dramatic maturation pathway that includes a particle size reduction of 100 A and an autocatalytic cleavage. Here we use cryo-EM and difference maps, computed at three time points following maturation initiation, to show that regions of N omegaV subunit folding are maturation dependent and occur at rates determined by their quasi-equivalent position in the capsid, explaining the unusual kinetics of the maturation cleavage. This study shows that folding is rapid and peptide chain self-cleavage occurs early for subunits adjacent to 3-fold and 5-fold icosahedral symmetry elements and that folding is slower in regions where molecular switches are required for the formation of the proper interfacial contacts. The results connect viral maturation to the well-studied assembly-dependent folding that occurs in the formation of cellular complexes.
Matsui
Tsutomu
T
Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Lander
Gabriel C
GC
Khayat
Reza
R
Johnson
John E
JE
D002213:Capsid; D020285:Cryoelectron Microscopy; D056890:Eukaryota / Q000821:virology; D007700:Kinetics; D010316:Particle Size; D012328:RNA Viruses / Q000502:physiology*; D019065:Virus Assembly