Role of Electrostatics in the assembly pathway of a single-stranded RNA virus

R.F. Garmann, M. Comas-Garcia, M.S.T. Koay, Jeroen Johannes Lambertus Maria Cornelissen, C.M. Knobler, W.M. Gelbart

Research output: Contribution to journalArticleAcademicpeer-review

52 Citations (Scopus)

Abstract

We have recently discovered (R. D. Cadena-Nava et al., J. Virol. 86:3318–3326, 2012, doi:10.1128/JVI.06566-11) that the in vitro packaging of RNA by the capsid protein (CP) of cowpea chlorotic mottle virus is optimal when there is a significant excess of CP, specifically that complete packaging of all of the RNA in solution requires sufficient CP to provide charge matching of the N-terminal positively charged arginine-rich motifs (ARMS) of the CPs with the negatively charged phosphate backbone of the RNA. We show here that packaging results from the initial formation of a charge-matched protocapsid consisting of RNA decorated by a disordered arrangement of CPs. This protocapsid reorganizes into the final, icosahedrally symmetric nucleocapsid by displacing the excess CPs from the RNA to the exterior surface of the emerging capsid through electrostatic attraction between the ARMs of the excess CP and the negative charge density of the capsid exterior. As a test of this scenario, we prepare CP mutants with extra and missing (relative to the wild type) cationic residues and show that a correspondingly smaller and larger excess, respectively, of CP is needed for complete packaging of RNA.
Original languageEnglish
Pages (from-to)10472-10481
Number of pages9
JournalJournal of virology
Volume88
Issue number18
DOIs
Publication statusPublished - 2014

Keywords

  • METIS-308472
  • IR-95089

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