Enhanced local symmetry interactions globally stabilize a mutant virus capsid that maintains infectivity and capsid dynamics

J Virol. 2006 Apr;80(7):3582-91. doi: 10.1128/JVI.80.7.3582-3591.2006.

Abstract

Structural transitions in viral capsids play a critical role in the virus life cycle, including assembly, disassembly, and release of the packaged nucleic acid. Cowpea chlorotic mottle virus (CCMV) undergoes a well-studied reversible structural expansion in vitro in which the capsid expands by 10%. The swollen form of the particle can be completely disassembled by increasing the salt concentration to 1 M. Remarkably, a single-residue mutant of the CCMV N-terminal arm, K42R, is not susceptible to dissociation in high salt (salt-stable CCMV [SS-CCMV]) and retains 70% of wild-type infectivity. We present the combined structural and biophysical basis for the chemical stability and viability of the SS-CCMV particles. A 2.7-A resolution crystal structure of the SS-CCMV capsid shows an addition of 660 new intersubunit interactions per particle at the center of the 20 hexameric capsomeres, which are a direct result of the K42R mutation. Protease-based mapping experiments of intact particles demonstrate that both the swollen and closed forms of the wild-type and SS-CCMV particles have highly dynamic N-terminal regions, yet the SS-CCMV particles are more resistant to degradation. Thus, the increase in SS-CCMV particle stability is a result of concentrated tethering of subunits at a local symmetry interface (i.e., quasi-sixfold axes) that does not interfere with the function of other key symmetry interfaces (i.e., fivefold, twofold, quasi-threefold axes). The result is a particle that is still dynamic but insensitive to high salt due to a new series of bonds that are resistant to high ionic strength and preserve the overall particle structure.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Bromovirus / chemistry*
  • Bromovirus / genetics
  • Bromovirus / isolation & purification
  • Bromovirus / metabolism
  • Bromovirus / ultrastructure
  • Capsid / chemistry*
  • Crystallography, X-Ray
  • Electrophoresis, Polyacrylamide Gel
  • Genome, Viral
  • Hydrogen Bonding
  • Hydrogen-Ion Concentration
  • Kinetics
  • Models, Molecular
  • Molecular Weight
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Structure, Secondary
  • RNA, Viral / analysis
  • Serine Endopeptidases / pharmacology
  • Sodium Chloride / pharmacology
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Trypsin / pharmacology
  • Water / chemistry

Substances

  • RNA, Viral
  • Water
  • Sodium Chloride
  • Serine Endopeptidases
  • glutamyl endopeptidase
  • Trypsin