Abstract

How are virus particles assembled and how do protein molecules move to build complicated structures, to send and receive signals, and to carry out many other biological functions? These are the broad questions of fundamental relevance to cancer biology that the proposed research on """"""""Virus Assembly and Protein Adaptability"""""""" will explore. Polyomavirus assembly will be investigated by electron microscopy of the aggregates formed in vitro from mutant and wild type recombinant capsid proteins, by three-dimensional image reconstruction from cryoelectron micrographs of distinctive polymorphic aggregates, and by X-ray diffraction analysis of the disordered nucleohistone core structure in crystalline virions. The structure of the polymorphic stacked-disk aggregate of tobacco mosaic virus (TMV) protein will be determined by a combination of high resolution electron microscopy and electron fiber diffraction form oriented frozen hydrated specimens. Information about this structure, that of the crystalline four layers disk aggregate and the intact TMV particle will be correlated with physiochemical data to characterize the electrostatic interactions that regulate the virus assembly. The colloidal ordering of the rod-shaped virus particles will be investigated by optical and small- angle X-ray diffraction methods. The conformational variations of crystalline insulin induced by changes in solvent conditions will be determined by high resolution X-ray crystallography; and the average solvent structure will be mapped from accurate low-resolution data. The coupling in the atomic movements of the insulin molecules under various conditions will be determined by analysis of diffuse X-ray scattering measurements. Models of the fluctuating conformational substates of the protein molecule will be refined to fit the experimental diffuse scattering data and energetic restraints. These structural studies will involve technological developments in the measurement of X-ray scattering data using synchrotron radiation and advanced X-ray area detectors; preparation of well oriented samples for high resolution electron fiber diffraction; and application of computer graphics methods to analyze and display structural data.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Unknown (R35)
Project #
5R35CA047439-11
Application #
2683479
Study Section
Special Emphasis Panel (SRC (88))
Program Officer
Cole, John S
Project Start
1988-05-06
Project End
2001-03-31
Budget Start
1998-04-01
Budget End
1999-03-31
Support Year
11
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
Florida State University
Department
Type
Organized Research Units
DUNS #
020520466
City
Tallahassee
State
FL
Country
United States
Zip Code
32306

  Publications

Adamek, D H; Guerrero, L; Blaber, M et al. (2005) Structural and energetic consequences of mutations in a solvated hydrophobic cavity. J Mol Biol 346:307-18
Diao, Jiasheng (2003) Crystallographic titration of cubic insulin crystals: pH affects GluB13 switching and sulfate binding. Acta Crystallogr D Biol Crystallogr 59:670-6
Twigg, P D; Parthasarathy, G; Guerrero, L et al. (2001) Disordered to ordered folding in the regulation of diphtheria toxin repressor activity. Proc Natl Acad Sci U S A 98:11259-64
King, C Y (2001) Supporting the structural basis of prion strains: induction and identification of [PSI] variants. J Mol Biol 307:1247-60
Diaz-Avalos, R; Caspar, D L (2000) Hyperstable stacked-disk structure of tobacco mosaic virus protein: electron cryomicroscopy image reconstruction related to atomic models. J Mol Biol 297:67-72
Yu, B; Blaber, M; Gronenborn, A M et al. (1999) Disordered water within a hydrophobic protein cavity visualized by x-ray crystallography. Proc Natl Acad Sci U S A 96:103-8
Twigg, P D; Wylie, G P; Wang, G et al. (1999) Expression and assignment of the 1H, 15N, and 13C resonances of the C-terminal domain of the diphtheria toxin repressor. J Biomol NMR 13:197-8
Wang, G; Wylie, G P; Twigg, P D et al. (1999) Solution structure and peptide binding studies of the C-terminal src homology 3-like domain of the diphtheria toxin repressor protein. Proc Natl Acad Sci U S A 96:6119-24
Zhou, G; Somasundaram, T; Blanc, E et al. (1998) Transition state structure of arginine kinase: implications for catalysis of bimolecular reactions. Proc Natl Acad Sci U S A 95:8449-54
Diaz-Avalos, R; Caspar, D L (1998) Structure of the stacked disk aggregate of tobacco mosaic virus protein. Biophys J 74:595-603

Showing the most recent 10 out of 33 publications