In vitro assembly of the øx174 procapsid from external scaffolding protein oligomers and early pentameric assembly intermediates

James E. Cherwa, Lindsey J. Organtini, Robert E. Ashley, Susan L. Hafenstein, Bentley A. Fane

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Bacteriophage øX174 morphogenesis requires two scaffolding proteins: an internal species, similar to those employed in other viral systems, and an external species, which is more typically associated with satellite viruses. The current model of øX174 assembly is based on structural and in vivo data. During morphogenesis, 240 copies of the external scaffolding protein mediate the association of 12 pentameric particles into procapsids. The hypothesized pentameric intermediate, the 12S* particle, contains 16 proteins: 5 copies each of the coat, spike and internal scaffolding proteins and 1 copy of the DNA pilot protein. Assembly naïve 12S* particles and external scaffolding oligomers, most likely tetramers, formed procapsid-like particles in vitro, suggesting that the 12S* particle is a bona fide assembly intermediate and validating the current model of procapsid morphogenesis. The in vitro system required a crowding agent, was influenced by the ratio of the reactants and was most likely driven by hydrophobic forces. While the system reported here shared some characteristics with other in vitro internal scaffolding protein-mediated systems, it displayed unique features. These features most likely reflect external scaffolding protein-mediated morphogenesis and the øX174 procapsid structure, in which external scaffolding-scaffolding protein interactions, as opposed to coat-coat protein interactions between pentamers, constitute the primary lattice-forming contacts.

Original languageEnglish (US)
Pages (from-to)387-396
Number of pages10
JournalJournal of Molecular Biology
Volume412
Issue number3
DOIs
StatePublished - Sep 23 2011

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Morphogenesis
Proteins
Satellite Viruses
In Vitro Techniques
Capsid Proteins
Bacteriophages
DNA

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Cite this

Cherwa, James E. ; Organtini, Lindsey J. ; Ashley, Robert E. ; Hafenstein, Susan L. ; Fane, Bentley A. / In vitro assembly of the øx174 procapsid from external scaffolding protein oligomers and early pentameric assembly intermediates. In: Journal of Molecular Biology. 2011 ; Vol. 412, No. 3. pp. 387-396.
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In vitro assembly of the øx174 procapsid from external scaffolding protein oligomers and early pentameric assembly intermediates. / Cherwa, James E.; Organtini, Lindsey J.; Ashley, Robert E.; Hafenstein, Susan L.; Fane, Bentley A.

In: Journal of Molecular Biology, Vol. 412, No. 3, 23.09.2011, p. 387-396.

Research output: Contribution to journalArticle

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