Eliminating the Requirement of an Essential Gene Product in an Already Very Small Virus: Scaffolding Protein B-free øX174, B-free

Min Chen, Asako Uchiyama, Bentley A. Fane

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

Unlike most viral assembly systems, two scaffolding proteins, B and D, mediate bacteriophage øX174 morphogenesis. The external scaffolding protein D is highly ordered in the atomic structure and proper function is very sensitive to mutation. In contrast, the internal scaffolding protein B is relatively unordered and extensive alterations do not eliminate function. Despite this genetic laxity, protein B is absolutely required for virus assembly. Thus, this system, with its complex arrangements of overlapping reading frames, can be regarded as an example of "irreducible complexity." To address the biochemical functions of a dual scaffolding protein system and the evolution of complexity, progressive and targeted genetic selections were employed to lessen and finally eliminate B protein-dependence. The biochemical and genetic bases of adaptation were characterized throughout the analysis that led to the sextuple mutant with a B-independent phenotype, as evaluated by plaque formation in wild-type cells. The primary adaptation appears to be the over-expression of a mutant external scaffolding protein. Progeny production was followed in lysis-resistant cells. The ability to produce infectious virions does not require all six mutations. However, the lag phase before progeny production is shortened as mutations accumulate. The results suggest that the primary function of the internal scaffolding protein may be to lower the critical concentration of the external scaffolding protein needed to nucleate procapsid formation. Moreover, they demonstrate a novel mechanism by which a stringently required gene product can be bypassed, even in a system encoding only eight strictly essential proteins.

Original languageEnglish (US)
Pages (from-to)308-314
Number of pages7
JournalJournal of Molecular Biology
Volume373
Issue number2
DOIs
StatePublished - Oct 19 2007

Keywords

  • evolution
  • irreducible complexity
  • scaffolding protein
  • virus assembly
  • øX174

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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