A research program will be undertaken to deepen our understanding of the molecular mechanisms underlying the regulation of chromatin structure and function. Chromatin, the complex of genomic DNA with histone and non-histone proteins, is the physiologically relevant form of all eukaryotic genomes. Once thought of as merely a packaging solution for the DNA, it is now accepted that chromatin serves as a dynamic signaling platform to regulate DNA access. Indeed, this research proposal is rooted in the idea that there exist heritable functional """"""""states"""""""" of chromatin, representing a fundamental regulatory mechanism that operates outside of the DNA itself, that help set the transcriptional potential of a gene locus. Mistakes made in establishing and maintaining these chromatin states, governed by chromatin remodeling activities, lead to the misregulation of genes with far-reaching implications for human biology and human disease. Building on some recent breakthroughs on the synthesis and application of what we call 'designer chromatin', we will integrate chemical, biochemical, biophysical and genetic tools for the purpose of obtaining a deeper understanding of the physicochemical principles underlying the regulation of chromatin function. The immediate focus will be on the role of histone ubiquitylation in chromatin biology (aims 1 and 2), however, new tools will also be developed that broaden the scope of this work significantly into other aspects of chromatin regulation and, importantly, that do so by greatly accelerating mechanistic biochemistry (aims 2 and 3). In the short term, this research is designed to reveal the molecular mechanism(s) by which histone ubiquitylation orchestrates downstream biochemistry on chromatin, an understanding that may aid the rational design of therapeutics directed at inhibiting enzymes such as the methyltransferase, hDot1L, for the treatment of leukemias. In the long term, the technologies that will be developed as part of this work, will have broad utility in the chromatin area. .

Public Health Relevance

DNA in eukaryotic cells is packaged into chromatin, the fundamental unit of which is the nucleosome in which DNA is wrapped around a spool of proteins called histones. The packaging of nucleosomes into higher-order chromatin structures is a key determinant of gene expression, and posttranslational modification of histone proteins is one way that chromatin structure is manipulated (1). The proposed research program is expected to reveal the molecular mechanism(s) by which one such modification, histone ubiquitylation, controls gene transcription through the recruitment and activation of effector proteins such as lysine methyltransferases. Such an understanding may aid the rational design of therapeutics directed at inhibiting these enzymes for the treatment of leukemias. Moreover, the high through-put technologies that will be developed as part of this program, will have broad utility in the chromatin biology area.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM107047-02
Application #
8724531
Study Section
Macromolecular Structure and Function C Study Section (MSFC)
Program Officer
Preusch, Peter
Project Start
2013-09-01
Project End
2017-04-30
Budget Start
2014-05-01
Budget End
2015-04-30
Support Year
2
Fiscal Year
2014
Total Cost
$299,888
Indirect Cost
$109,888
Name
Princeton University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
002484665
City
Princeton
State
NJ
Country
United States
Zip Code
08544
David, Yael; Muir, Tom W (2017) Emerging Chemistry Strategies for Engineering Native Chromatin. J Am Chem Soc 139:9090-9096
Debelouchina, Galia T; Gerecht, Karola; Muir, Tom W (2017) Ubiquitin utilizes an acidic surface patch to alter chromatin structure. Nat Chem Biol 13:105-110
Dann, Geoffrey P; Liszczak, Glen P; Bagert, John D et al. (2017) ISWI chromatin remodellers sense nucleosome modifications to determine substrate preference. Nature 548:607-611
Liszczak, Glen P; Brown, Zachary Z; Kim, Samuel H et al. (2017) Genomic targeting of epigenetic probes using a chemically tailored Cas9 system. Proc Natl Acad Sci U S A 114:681-686
Lu, Chao; Jain, Siddhant U; Hoelper, Dominik et al. (2016) Histone H3K36 mutations promote sarcomagenesis through altered histone methylation landscape. Science 352:844-9
Müller, Manuel M; Fierz, Beat; Bittova, Lenka et al. (2016) A two-state activation mechanism controls the histone methyltransferase Suv39h1. Nat Chem Biol 12:188-93
Zhou, Linjiao; Holt, Matthew T; Ohashi, Nami et al. (2016) Evidence that ubiquitylated H2B corrals hDot1L on the nucleosomal surface to induce H3K79 methylation. Nat Commun 7:10589
Holt, Matthew T; David, Yael; Pollock, Sam et al. (2015) Identification of a functional hotspot on ubiquitin required for stimulation of methyltransferase activity on chromatin. Proc Natl Acad Sci U S A 112:10365-70
Holt, Matthew; Muir, Tom (2015) Application of the protein semisynthesis strategy to the generation of modified chromatin. Annu Rev Biochem 84:265-90
Müller, Manuel M; Muir, Tom W (2015) Histones: at the crossroads of peptide and protein chemistry. Chem Rev 115:2296-349

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