Abstract

The overall goal of this work is to elucidate the interaction of linker histones with chromatin. Eight different studies are proposed in four groups. Experiments 1 & 2 use a DNA cleavage assay to determine the location and orientation of GH5 and its C- terminal tail bound to nucleosomes. Experiment 3 measures the Kd for nucleosome binding. Experiments 4 & 5 examine the roles of particular residues in nucleosome binding and in cooperative binding to DNA. Experiment 6 addresses sequence-preferences in DNA binding. Experiments 7 & 8 are proposed studies of protein-DNA co-crystals to elucidate the binding of GH5 and its C-terminal tail.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM042796-07
Application #
2022323
Study Section
Special Emphasis Panel (ZRG3-BBCB (01))
Project Start
1989-07-01
Project End
1999-11-30
Budget Start
1996-12-01
Budget End
1997-11-30
Support Year
7
Fiscal Year
1997
Total Cost
Indirect Cost

  Institution

Name
University of Utah
Department
Biochemistry
Type
Schools of Medicine
DUNS #
City
Salt Lake City
State
UT
Country
United States
Zip Code
84112

  Publications

Angus-Hill, M L; Dutnall, R N; Tafrov, S T et al. (1999) Crystal structure of the histone acetyltransferase Hpa2: A tetrameric member of the Gcn5-related N-acetyltransferase superfamily. J Mol Biol 294:1311-25
Dutnall, R N; Tafrov, S T; Sternglanz, R et al. (1998) Structure of the histone acetyltransferase Hat1: a paradigm for the GCN5-related N-acetyltransferase superfamily. Cell 94:427-38
Ramakrishnan, V (1997) Histone H1 and chromatin higher-order structure. Crit Rev Eukaryot Gene Expr 7:215-30
Ramakrishnan, V (1997) Histone structure and the organization of the nucleosome. Annu Rev Biophys Biomol Struct 26:83-112
Ramakrishnan, V; Biou, V (1997) Treatment of multiwavelength anomalous diffraction data as a special case of multiple isomorphous replacement. Methods Enzymol 276:538-57
Goytisolo, F A; Gerchman, S E; Yu, X et al. (1996) Identification of two DNA-binding sites on the globular domain of histone H5. EMBO J 15:3421-9
Hamiche, A; Schultz, P; Ramakrishnan, V et al. (1996) Linker histone-dependent DNA structure in linear mononucleosomes. J Mol Biol 257:30-42
Graziano, V; Gerchman, S E; Schneider, D K et al. (1996) Neutron scattering studies on chromatin higher-order structure. Basic Life Sci 64:127-36
Cerf, C; Lippens, G; Ramakrishnan, V et al. (1994) Homo- and heteronuclear two-dimensional NMR studies of the globular domain of histone H1: full assignment, tertiary structure, and comparison with the globular domain of histone H5. Biochemistry 33:11079-86
Gerchman, S E; Graziano, V; Ramakrishnan, V (1994) Expression of chicken linker histones in E. coli: sources of problems and methods for overcoming some of the difficulties. Protein Expr Purif 5:242-51

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