title: Analysis of a salt stable mutant of cowpea chlorotic mottle virus.
abstract: An understanding of virion assembly and disassembly requires a detailed understanding of the protein-protein and protein-nucleic acid interactions which stabilize the virion. We have characterized a mutant of cowpea chlorotic mottle virus (CCMV) that is altered in virion stability. The mutant virions resist disassembly in 1.0 M NaCl, pH 7.5, whereas the wild-type virions completely disassociate into RNA and capsid protein components. Sequence analysis of the mutant coat protein gene identified a single A to G nucleotide change at position 1484 of RNA 3 (position 134 of RNA 4), which results in a lysine to arginine change at position 42 of the coat protein. Introduction of the K42R mutation into wild-type CCMV coat protein results in a salt stable virion phenotype. Likewise, expression of the K42R mutant coat protein in Escherichia coli followed by in vitro assembly produces virions that exhibit the salt stable phenotype. Analysis of this mutation demonstrates how a single amino acid change in the primary structure of the coat protein leads to tertiary interactions which stabilize the virion.
authors.affiliation: Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA.
authors.forename: J M
authors.initials: JM
authors.lastname: Fox
authors.forename: X
authors.initials: X
authors.lastname: Zhao
authors.forename: J A
authors.initials: JA
authors.lastname: Speir
authors.forename: M J
authors.initials: MJ
authors.lastname: Young
mesh_terms: D017795:Bromovirus / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D002213:Capsid / Q000235:genetics; D018076:DNA, Complementary; D004352:Drug Resistance, Microbial / Q000235:genetics; D004926:Escherichia coli; D016297:Mutagenesis, Site-Directed; D010641:Phenotype; D011994:Recombinant Proteins / Q000235:genetics; D012965:Sodium Chloride / Q000494:pharmacology*; D014771:Virion; D019065:Virus Assembly
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D012965:Sodium Chloride