title: The bipolar filaments formed by herpes simplex virus type 1 SSB/recombination protein (ICP8) suggest a mechanism for DNA annealing.
abstract: Herpes simplex virus type 1 encodes a multifunctional protein, ICP8, which serves both as a single-strand binding protein and as a recombinase, catalyzing reactions involved in replication and recombination of the viral genome. In the presence of divalent ions and at low temperature, previous electron microscopic studies showed that ICP8 will form long left-handed helical filaments. Here, electron microscopic image reconstruction reveals that the filaments are bipolar, with an asymmetric unit containing two subunits of ICP8 that constitute a symmetrical dimer. This organization of the filament has been confirmed using scanning transmission electron microscopy. The pitch of the filaments is approximately 250 A, with approximately 6.2 dimers per turn. Docking of a crystal structure of ICP8 into the reconstructed filament shows that the C-terminal domain of ICP8, attached to the body of the subunit by a flexible linker containing approximately 10 residues, is packed into a pocket in the body of a neighboring subunit in the crystal in a similar manner as in the filament. However, the interactions between the large N-terminal domains are quite different in the filament from that observed in the crystal. A previously proposed model for ICP8 binding single-stranded DNA (ssDNA), based upon the crystal structure, leads to a model for a continuous strand of ssDNA near the filament axis. The bipolar nature of the ICP8 filaments means that a second strand of ssDNA would be running through this filament in the opposite orientation, and this provides a potential mechanism for how ICP8 anneals complementary ssDNA into double-stranded DNA, where each strand runs in opposite directions.
authors.affiliation: Lineberger Comprehensive Cancer Center, Department Microbiology and Immunology, University of North Carolina at Chapel Hill, 27517, USA.
authors.forename: Alexander M
authors.initials: AM
authors.lastname: Makhov
authors.forename: Anindito
authors.initials: A
authors.lastname: Sen
authors.forename: Xiong
authors.initials: X
authors.lastname: Yu
authors.forename: Martha N
authors.initials: MN
authors.lastname: Simon
authors.forename: Jack D
authors.initials: JD
authors.lastname: Griffith
authors.forename: Edward H
authors.initials: EH
authors.lastname: Egelman
mesh_terms: D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D018259:Herpesvirus 1, Human / Q000502:physiology*; D007091:Image Processing, Computer-Assisted; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D055503:Protein Multimerization; D011995:Recombination, Genetic; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*
chemical_list: D004268:DNA-Binding Proteins; C045462:ICP8 protein, Simplexvirus; D014764:Viral Proteins; D004247:DNA