The long-term objectives of this program are to study the structure of nucleosomes and other key macromolecular constituents of genetic material. These macromolecules are elements in the primary functions of chromatin: transcription and regulation, replication and DNA packaging. In order to understand the function of these molecules, several biophysical and biochemical techniques will be emphasized, with the primary effort consisting of (1) x-ray crystallography of nucleosomes and (2) studies of the macromolecular components of genetic material in solution using the technique of small-angle neutron scattering (SANS) and small angle x-raly scattering (SAXS). The x-ray crystallography program will consist of the study and interpretation of a low resolution 6-8 Angstroms electron density map of the nucleosome, development of nucleosome crystals which diffract to medium or high resolution (about 3 Angstroms) to be accomplished by reconstituting nucleosomes from the purified histones and a DNA component of defined length and sequnce, and determination of the structure by crystallographic methods. The small-angle scattering program will concentrate on materials closely associated with potentially active genes, such as HMG - nucleosomes, """"""""active nucleosomes"""""""", and other chromatin species such as dinucleosomes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM029818-04
Application #
3277483
Study Section
Biophysics and Biophysical Chemistry A Study Section (BBCA)
Project Start
1982-01-01
Project End
1987-12-31
Budget Start
1985-01-01
Budget End
1985-12-31
Support Year
4
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Tennessee Knoxville
Department
Type
Graduate Schools
DUNS #
City
Knoxville
State
TN
Country
United States
Zip Code
37996
Hanson, B Leif; Bunick, Gerard J (2007) Annealing macromolecular crystals. Methods Mol Biol 364:31-42
Katz, Amy K; Li, Xinmin; Carrell, H L et al. (2006) Locating active-site hydrogen atoms in D-xylose isomerase: time-of-flight neutron diffraction. Proc Natl Acad Sci U S A 103:8342-7
Hanson, B Leif; Alexander, Chad; Harp, Joel M et al. (2004) Preparation and crystallization of nucleosome core particle. Methods Enzymol 375:44-62
Hanson, B Leif; Langan, Paul; Katz, Amy K et al. (2004) A preliminary time-of-flight neutron diffraction study of Streptomyces rubiginosus D-xylose isomerase. Acta Crystallogr D Biol Crystallogr 60:241-9
Bunick, Christopher G; Nelson, Melanie R; Mangahas, Sheryll et al. (2004) Designing sequence to control protein function in an EF-hand protein. J Am Chem Soc 126:5990-8
Hanson, B Leif; Schall, Constance A; Bunick, Gerard J (2003) New techniques in macromolecular cryocrystallography: macromolecular crystal annealing and cryogenic helium. J Struct Biol 142:77-87
Hanson, B Leif; Harp, Joel M; Bunick, Gerard J (2003) The well-tempered protein crystal: annealing macromolecular crystals. Methods Enzymol 368:217-35
Harp, J M; Hanson, B L; Timm, D E et al. (2000) Asymmetries in the nucleosome core particle at 2.5 A resolution. Acta Crystallogr D Biol Crystallogr 56:1513-34
Timm, D E; Mueller, H A; Bhanumoorthy, P et al. (1999) Crystal structure and mechanism of a carbon-carbon bond hydrolase. Structure 7:1023-33
Harp, J M; Hanson, B L; Timm, D E et al. (1999) Macromolecular crystal annealing: evaluation of techniques and variables. Acta Crystallogr D Biol Crystallogr 55:1329-34

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