Chromatin plays two essential, yet by their very nature mutually exclusive roles. On one hand, chromatin is responsible for the extreme degree of DNA compaction that allows the eukaryotic genome to fit into the confines of the nucleus. On the other hand, it must permit the local unraveling of DNA to grant access of the cellular machinery to the genome. Activities that mediate the inter- conversion between various levels of compacted chromatin states are key regulators of all processes requiring access to genomic DNA. Mechanistic and structural insight into these biologically relevant processes is limited. Here, the role of the nucleosome assembly protein 1 (NAP1) family of histone chaperones in maintaining and modulating chromatin structure and fluidity will be investigated.
Aims 1 and 2 address the structural and molecular basis for chaperone interaction with histones, using x-ray diffraction as well as biochemical and in vivo approaches. The mechanism and biological role of NAP1-mediated histone removal and histone exchange will be studied by rigorous kinetic analysis combined with in vivo examination of chromatin structure (aim 3). Finally, the novel hypothesis that a posttranslational modification regulates NAP1 activity in metazoans will be put to test in aim 4. The strength of this proposal lies in a combination of a broad spectrum of structural, biophysical, biochemical and genetic approaches to address the highly significant question of how compacted DNA is made available to the cellular machinery.

Public Health Relevance

The molecular mechanism and biological relevance of histone chaperone-mediated chromatin structure modulation will be investigated, using a combination of structural, biophysical, and genetic approaches.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM067777-08S1
Application #
8397737
Study Section
Special Emphasis Panel (ZRG1-BCMB-B (02))
Program Officer
Preusch, Peter C
Project Start
2003-05-01
Project End
2012-08-31
Budget Start
2010-12-01
Budget End
2012-08-31
Support Year
8
Fiscal Year
2012
Total Cost
$61,937
Indirect Cost
$19,441
Name
Colorado State University-Fort Collins
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
785979618
City
Fort Collins
State
CO
Country
United States
Zip Code
80523
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White, Alison E; Hieb, Aaron R; Luger, Karolin (2016) A quantitative investigation of linker histone interactions with nucleosomes and chromatin. Sci Rep 6:19122
Prasad, Rashmi; D'Arcy, Sheena; Hada, Arjan et al. (2016) Coordinated Action of Nap1 and RSC in Disassembly of Tandem Nucleosomes. Mol Cell Biol 36:2262-71
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D'Arcy, Sheena; Martin, Kyle W; Panchenko, Tanya et al. (2013) Chaperone Nap1 shields histone surfaces used in a nucleosome and can put H2A-H2B in an unconventional tetrameric form. Mol Cell 51:662-77
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