title: A conserved leucine that constricts the pore through the capsid fivefold cylinder plays a central role in parvoviral infection.
abstract: The atomic structure of the DNA-containing T = 1 particle of the parvovirus minute virus of mice (MVM) reveals cylindrical projections at each fivefold symmetry axis, each containing an 8 Angstrom pore through which runs 10 amino acids of a single VP2 N-terminus. The tightest constriction of this pore is formed at its inner end by the juxtaposition of leucine side chains from position 172 of five independent VP2 molecules. To test whether L172 modulates the extrusion of VP N-termini, we constructed and analyzed a complete set of amino acid substitution mutants at this highly conserved residue. All but one mutant produced DNA-containing virions, but only two, L172V and L172I, were infectious, the others being blocked for viral entry. Several mutants were significantly defective for assembly at 39 degrees C, but not at 32 degrees C. L172W significantly impaired genome encapsidation, indicating that the fivefold cylinder may also be the DNA packaging portal. Although tryptic cleavage of the VP2 N-terminus was not affected for the mutants, VP1 was degraded during proteolysis of mutant, but not wild-type, virions.
authors.affiliation: Department of Laboratory Medicine, Yale University School of Medicine, New Haven, CT 06510, USA.
authors.forename: Glen A
authors.initials: GA
authors.lastname: Farr
authors.forename: Peter
authors.initials: P
authors.lastname: Tattersall
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D002213:Capsid / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D017124:Conserved Sequence; D015967:Gene Expression Regulation, Viral; D007930:Leucine; D008915:Minute Virus of Mice / Q000737:chemistry / Q000472:pathogenicity*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D013696:Temperature; D014771:Virion / Q000378:metabolism / Q000648:ultrastructure; D019065:Virus Assembly
chemical_list: D036022:Capsid Proteins; D007930:Leucine