The role of scaffolding proteins in the assembly of the small, single-stranded DNA virus phiX174.

Academic Article

Abstract

  • An empty precursor particle called the procapsid is formed during assembly of the single-stranded DNA bacteriophage phiX174. Assembly of the phiX174 procapsid requires the presence of the two scaffolding proteins, D and B, which are structural components of the procapsid, but are not found in the mature virion. The X-ray crystallographic structure of a "closed" procapsid particle has been determined to 3.5 A resolution. This structure has an external scaffold made from 240 copies of protein D, 60 copies of the internally located B protein, and contains 60 copies of each of the viral structural proteins F and G, which comprise the shell and the 5-fold spikes, respectively. The F capsid protein has a similar conformation to that seen in the mature virion, and differs from the previously determined 25 A resolution electron microscopic reconstruction of the "open" procapsid, in which the F protein has a different conformation. The D scaffolding protein has a predominantly alpha-helical fold and displays remarkable conformational variability. We report here an improved and refined structure of the closed procapsid and describe in some detail the differences between the four independent D scaffolding proteins per icosahedral asymmetric unit, as well as their interaction with the F capsid protein. We re-analyze and correct the comparison of the closed procapsid with the previously determined cryo-electron microscopic image reconstruction of the open procapsid and discuss the major structural rearrangements that must occur during assembly. A model is proposed in which the D proteins direct the assembly process by sequential binding and conformational switching.
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    Published In

    Keywords

  • Amino Acid Sequence, Bacteriophage phi X 174, Capsid, Crystallography, X-Ray, Microscopy, Electron, Models, Molecular, Molecular Sequence Data, Protein Conformation, Virus Assembly
  • Digital Object Identifier (doi)

    Author List

  • Dokland T; Bernal RA; Burch A; Pletnev S; Fane BA; Rossmann MG
  • Start Page

  • 595
  • End Page

  • 608
  • Volume

  • 288
  • Issue

  • 4