Functional relevance of amino acid residues involved in interactions with ordered nucleic acid in a spherical virus.
In the spherical virion of the parvovirus minute virus of mice, several amino acid side chains of the capsid were previously found to be involved in interactions with the viral single-stranded DNA molecule. We have individually truncated by mutation to alanine many (ten) of these side chains and analyzed the effects on capsid assembly, stability and conformation, viral DNA encapsidation, and virion infectivity. Mutation of residues Tyr-270, Asp-273, or Asp-474 led to a drastic reduction in infectivity. Mutant Y270A was defective in capsid assembly; mutant D273A formed stable capsids, but it was essentially unable to encapsidate the viral DNA or to externalize the N terminus of the capsid protein VP2, a connected conformational event. Mutation of residues Asp-58, Trp-60, Asn-183, Thr-267, or Lys-471 led to a moderate reduction in infectivity. None of these mutations had an effect on capsid assembly or stability, or on the DNA encapsidation process. However, those five mutant virions were substantially less stable than the parental virion in thermal inactivation assays. The results with this model spherical virus indicate that several capsid residues that are found to be involved in polar interactions or multiple hydrophobic contacts with the viral DNA molecule contribute to preserving the active conformation of the infectious viral particle. Their effect appears to be mediated by the non-covalent interactions they establish with the viral DNA. In addition, at least one acidic residue at each DNA-binding region is needed for DNA packaging.
Reguera
Juan
J
Centro de Biología Molecular "Severo Ochoa" (Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid), Universidad Autónoma de Madrid, Cantoblanco 28049 Madrid, Spain.
Grueso
Esther
E
Carreira
Aura
A
Sánchez-Martínez
Cristina
C
Almendral
José M
JM
Mateu
Mauricio G
MG
D000596:Amino Acids / Q000737:chemistry / Q000235:genetics; D000818:Animals; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics; D002460:Cell Line; D004252:DNA Mutational Analysis / Q000379:methods; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009696:Nucleic Acids / Q000737:chemistry / Q000235:genetics; D017992:Parvovirus / Q000737:chemistry* / Q000235:genetics*; D014771:Virion / Q000737:chemistry* / Q000235:genetics*
D000596:Amino Acids; D036022:Capsid Proteins; D009696:Nucleic Acids