My part of this grant application concerns structural studies of small, single-stranded, RNA viruses and their structural components using cryo=electron microscopy (cryoEM) and X-ray crystallography. The basic concept, which we and others have been exploring in the last decade, is that, whereas larger biological units are generally unlikely to crystallize, their three-dimensional structures can frequently be determined to 9 A resolution or better by cryoEM. At the same time, the atomic structures of some of the component proteins can be determined by X-ray crystallography to near atomic resolution (probably to better than 3 A resolution). These two types of results can be combined to give """"""""quasi-atomic"""""""" structures of whole viruses or virus complexes. Although more than half of our effort will be directed to a continuation of our previous investigation of alphaviruses, in particular of Sindbis virus, I plan to extend our efforts to the study of larger and probably less symmetric, enveloped, RNA viruses, including rubella virus (another genus of alphavirus for which it is not clear whether the virions have reproducible icosahedral symmetry), coronaviruses, and possibly also measles virus (an orthomyxovirus) and Hantaan virus (a bunyavirus). The work on alphaviruses will include the determination of the still structurally unknown E2 glycoprotein; extension of the cryoEM map of Sindbis virus to possibly 6 A resolution; study of the immature, pre-maturation, E2-E3 uncleaved virus; analysis of the RNA structure in the core; examinations of the virus bound with cellular receptor molecules and neutralizing antibodies; and further attempts to crystallize Sindbis virus. Work on other ssRNA viruses will need to go hand-in-hand with the development of methods to determine whole virus structure when the virions are asymmetric and pleomorphic, albeit similar in localized surface features. At the same time it will be necessary to start exploring the crystal structures of the component structural proteins. The major emphasis and the major challenge will be the study of the pleomorphic, non-symmetrical enveloped viruses. Tentatively, we plan to use cryoEM tomography to identify regions of local symmetry in the viral membrane and then to average (if appropriate) these regions between different particles. Alternatively, it might be possible to search cryoEM images of a pleomorphic virus for common patterns. These might represent different orientations of a locally recurring structure.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
5P01AI045976-10
Application #
7670255
Study Section
Special Emphasis Panel (ZAI1)
Project Start
Project End
Budget Start
2008-08-01
Budget End
2009-07-31
Support Year
10
Fiscal Year
2008
Total Cost
$316,464
Indirect Cost
Name
Purdue University
Department
Type
DUNS #
072051394
City
West Lafayette
State
IN
Country
United States
Zip Code
47907
Parent, Kristin N; Gilcrease, Eddie B; Casjens, Sherwood R et al. (2012) Structural evolution of the P22-like phages: comparison of Sf6 and P22 procapsid and virion architectures. Virology 427:177-88
Brodin, Jeffrey D; Ambroggio, X I; Tang, Chunyan et al. (2012) Metal-directed, chemically tunable assembly of one-, two- and three-dimensional crystalline protein arrays. Nat Chem 4:375-82
Kwon, Soonshin; Chen, Zack C Y; Kim, Ji-Hun et al. (2012) Misfit-guided self-organization of anticorrelated Ge quantum dot arrays on Si nanowires. Nano Lett 12:4757-62
Khadka, Sudip; Vangeloff, Abbey D; Zhang, Chaoying et al. (2011) A physical interaction network of dengue virus and human proteins. Mol Cell Proteomics 10:M111.012187
Tang, Jinghua; Lander, Gabriel C; Olia, Adam S et al. (2011) Peering down the barrel of a bacteriophage portal: the genome packaging and release valve in p22. Structure 19:496-502
Tang, Jinghua; Jose, Joyce; Chipman, Paul et al. (2011) Molecular links between the E2 envelope glycoprotein and nucleocapsid core in Sindbis virus. J Mol Biol 414:442-59
Ku, Ti-Hsuan; Chien, Miao-Ping; Thompson, Matthew P et al. (2011) Controlling and switching the morphology of micellar nanoparticles with enzymes. J Am Chem Soc 133:8392-5
Rossmann, Michael G; Battisti, Anthony J; Plevka, Pavel (2011) Future prospects. Adv Protein Chem Struct Biol 82:101-21
Shen, Peter S; Enderlein, Dirk; Nelson, Christian D S et al. (2011) The structure of avian polyomavirus reveals variably sized capsids, non-conserved inter-capsomere interactions, and a possible location of the minor capsid protein VP4. Virology 411:142-52
Yan, Xiaodong; Parent, Kristin N; Goodman, Russell P et al. (2011) Virion structure of baboon reovirus, a fusogenic orthoreovirus that lacks an adhesion fiber. J Virol 85:7483-95

Showing the most recent 10 out of 52 publications