title: Interaction with capsid protein alters RNA structure and the pathway for in vitro assembly of cowpea chlorotic mottle virus.
abstract: Viruses use sophisticated mechanisms to allow the specific packaging of their genome over that of host nucleic acids. We examined the in vitro assembly of the Cowpea chlorotic mottle virus (CCMV) and observed that assembly with viral RNA follows two different mechanisms. Initially, CCMV capsid protein (CP) dimers bind RNA with low cooperativity and form virus-like particles of 90 CP dimers and one copy of RNA. Longer incubation reveals a different assembly path. At a stoichiometry of about ten CP dimers per RNA, the CP slowly folds the RNA into a compact structure that can be bound with high cooperativity by additional CP dimers. This folding process is exclusively a function of CP quaternary structure and is independent of RNA sequence. CP-induced folding is distinct from RNA folding that depends on base-pairing to stabilize tertiary structure. We hypothesize that specific encapsidation of viral RNA is a three-step process: specific binding by a few copies of CP, RNA folding, and then cooperative binding of CP to the "labeled" nucleoprotein complex. This mechanism, observed in a plant virus, may be applicable to other viruses that do not halt synthesis of host nucleic acid, including HIV.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73190, USA.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Johnson
authors.forename: Deborah A
authors.initials: DA
authors.lastname: Willits
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Young
authors.forename: Adam
authors.initials: A
authors.lastname: Zlotnick
mesh_terms: D017795:Bromovirus / Q000254:growth & development / Q000502:physiology*; D036022:Capsid Proteins / Q000032:analysis / Q000378:metabolism*; D018076:DNA, Complementary; D019281:Dimerization; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D010944:Plants / Q000821:virology*; D012367:RNA, Viral / Q000378:metabolism*; D015398:Signal Transduction; D014764:Viral Proteins / Q000378:metabolism*; D019065:Virus Assembly
chemical_list: D036022:Capsid Proteins; D018076:DNA, Complementary; D012367:RNA, Viral; D014764:Viral Proteins