): Our long-term goal is to determine the atomic structures of herpes simplex virus type 1 (HSV-1) particles at different maturational stages and to elucidate their interactions with cellular factors. An intact HSV-1 virion is spherical with a diameter of about 2000 A and consists of several compartments: glycoproteins, membrane, tegument layer, capsid and dsDNA. Because of its size and complexity, we plan to employ a combination of electron cryomicroscopic, x-ray crystallographic, computational, biochemical, and genetic approaches to study the entire virion, the capsid and the individual capsid proteins. The capsid has a diameter of 1250 A and has a T=16 icosahedral lattice symmetry. The capsid shell has a total mass of 0.2 billion daltons and is made up of four different proteins (VP5, VP19C, VP23 and VP26) with molecular masses of 149, 50, 34 and 12 kDa respectively. This proposal will be carried out in collaboration with Frazer Rixon, a molecular virologist and Florante Quiocho, a crystallographer and will utilise established resources in the form of intact and mutant virions and different types of naturally occurring and recombinant capsid particles. We have six specific aims. The first is to obtain the crystal structures of individual molecular components of the capsid shell and to dock them into the intermediate resolution (6-8 A) capsid structure determined by electron cryomicroscopy. The second is to extend the structural study of the entire B capsid shell towards 6 A resolution by electron cryomicroscopy. The third is to integrate data from a variety of crystallographic, computational and labeling experiments to interpret the 6 A capsid map in terms of secondary structure elements, specific residue locations and possible folds. The fourth is to follow the process of capsid maturation by determining the structures of a number of intermediates from procapsid to mature virion. The fifth is to determine the identity of the icosahedrally-bound tegument protein that we have recently observed and also to characterize its structure at a higher resolution than at present. The sixth is to explore the possibilities for studying the intact capsid at atomic resolution by crystallography and electron cryomicroscopy. This will be the largest icosahedral particle to be studied in such detail. The outcome of our investigations will provide insights on the structural assembly of this important human pathogen and also advance the structural approaches for studying large macromolecular machines.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI038469-08
Application #
6631808
Study Section
Biophysical Chemistry Study Section (BBCB)
Program Officer
Beisel, Christopher E
Project Start
1995-09-30
Project End
2006-02-28
Budget Start
2003-03-01
Budget End
2004-02-29
Support Year
8
Fiscal Year
2003
Total Cost
$376,250
Indirect Cost
Name
Baylor College of Medicine
Department
Biochemistry
Type
Schools of Medicine
DUNS #
051113330
City
Houston
State
TX
Country
United States
Zip Code
77030
Chang, Juan T; Schmid, Michael F; Rixon, Frazer J et al. (2007) Electron cryotomography reveals the portal in the herpesvirus capsid. J Virol 81:2065-8
Jiang, Wen; Chiu, Wah (2007) Cryoelectron microscopy of icosahedral virus particles. Methods Mol Biol 369:345-63
Jiang, Wen; Chang, Juan; Jakana, Joanita et al. (2006) Structure of epsilon15 bacteriophage reveals genome organization and DNA packaging/injection apparatus. Nature 439:612-6
Chang, Juan; Weigele, Peter; King, Jonathan et al. (2006) Cryo-EM asymmetric reconstruction of bacteriophage P22 reveals organization of its DNA packaging and infecting machinery. Structure 14:1073-82
Baker, Matthew L; Jiang, Wen; Wedemeyer, William J et al. (2006) Ab initio modeling of the herpesvirus VP26 core domain assessed by CryoEM density. PLoS Comput Biol 2:e146
Bowman, Brian R; Welschhans, Robert L; Jayaram, Hariharan et al. (2006) Structural characterization of the UL25 DNA-packaging protein from herpes simplex virus type 1. J Virol 80:2309-17
Paredes, A; Weaver, S; Watowich, S et al. (2005) Structural biology of old world and new world alphaviruses. Arch Virol Suppl :179-85
Chiu, Wah; Baker, Matthew L; Jiang, Wen et al. (2005) Electron cryomicroscopy of biological machines at subnanometer resolution. Structure 13:363-72
Saad, Ali Samir (2005) Orientation determination by wavelets matching for 3D reconstruction of very noisy electron microscopic virus images. BMC Struct Biol 5:5
Booth, Christopher R; Jiang, Wen; Baker, Matthew L et al. (2004) A 9 angstroms single particle reconstruction from CCD captured images on a 200 kV electron cryomicroscope. J Struct Biol 147:116-27

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