title: Visualization of uncorrelated, tandem symmetry mismatches in the internal genome packaging apparatus of bacteriophage T7.
abstract: Motor-driven packaging of a dsDNA genome into a preformed protein capsid through a unique portal vertex is essential in the life cycle of a large number of dsDNA viruses. We have used single-particle electron cryomicroscopy to study the multilayer structure of the portal vertex of the bacteriophage T7 procapsid, the recipient of T7 DNA in packaging. A focused asymmetric reconstruction method was developed and applied to selectively resolve neighboring pairs of symmetry-mismatched layers of the portal vertex. However, structural features in all layers of the multilayer portal vertex could not be resolved simultaneously. Our results imply that layers with mismatched symmetries can join together in several different relative orientations, and that orientations at different interfaces assort independently to produce structural isomers, a process that we call combinatorial assembly isomerism. This isomerism explains rotational smearing in previously reported asymmetric reconstructions of the portal vertex of T7 and other bacteriophages. Combinatorial assembly isomerism may represent a new regime of structural biology in which globally varying structures assemble from a common set of components. Our reconstructions collectively validate previously proposed symmetries, compositions, and sequential order of T7 portal vertex layers, resolving in tandem the 5-fold gene product 10 (gp10) shell, 12-fold gp8 portal ring, and an internal core stack consisting of 12-fold gp14 adaptor ring, 8-fold bowl-shaped gp15, and 4-fold gp16 tip. We also found a small tilt of the core stack relative to the icosahedral fivefold axis and propose that this tilt assists DNA spooling without tangling during packaging.
authors.affiliation: Markey Center for Structural Biology, Department of Biological Sciences, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: Fei
authors.initials: F
authors.lastname: Guo
authors.forename: Zheng
authors.initials: Z
authors.lastname: Liu
authors.forename: Frank
authors.initials: F
authors.lastname: Vago
authors.forename: Yue
authors.initials: Y
authors.lastname: Ren
authors.forename: Weimin
authors.initials: W
authors.lastname: Wu
authors.forename: Elena T
authors.initials: ET
authors.lastname: Wright
authors.forename: Philip
authors.initials: P
authors.lastname: Serwer
authors.forename: Wen
authors.initials: W
authors.lastname: Jiang
mesh_terms: D017123:Bacteriophage T7 / Q000235:genetics*; D002213:Capsid / Q000648:ultrastructure*; D036022:Capsid Proteins / Q000235:genetics*; D020285:Cryoelectron Microscopy; D004926:Escherichia coli; D016679:Genome, Viral / Q000235:genetics*; D021621:Imaging, Three-Dimensional; D008958:Models, Molecular; D019065:Virus Assembly / Q000235:genetics*
chemical_list: D036022:Capsid Proteins