Chromatin was first defined by Flemming in 1882 as the 'substance in the cell nucleus which is readily Stained'that "is retractile to digestion'. The term stuck, and now is taken to refer to DNA (the readily stained component) and the associated proteins that make it retractile to digestion. Over the past two decades, it has become apparent that the associated proteins play a key role in regulating transcription, replication and recombination ofthe DNA. The basic structural component of chromatin, the nucleosome, which was treated for years as a 'dumb'building block, has been shown to be dynamic and regulated at several levels. Nucleosomes can be evicted or shifted in position by ATP-dependent remodeling proteins, opening up DNA sequences. They can be covalently modified in different ways, allowing differential abilities to interact with the regulatory machinery. The purpose ofthe experiments described in this application is to characterize regulatory proteins that modulate chromatin structure to effect gene regulation. The three Aims examine three separate complexes involved in these processes, all of which are human homologs of proteins originally identified in screens done in Drosophila for mutations that impact developmental progression. These screens identified the Polycomb-Group (PcG) of genes, responsible for maintaining repression of master regulatory genes during development, and the trithorax-Group (trxG), isolated as suppressor of a Polycomb mutant.
Aims 1 and 2 examine two mammalian ATP-dependent remodeling proteins in the trxG family and Aim 3 examines a mammalian protein in the PcG family.
Aim 1 will determine the crystal structure of the ATPase domain of BRG1;
Aim 2 will examine the impact of human disease mutations of function of the CHD7-ATP-dependent r

Public Health Relevance

The CHD7 gene is mutated in 90% of human patients that have CHARGE syndrome, a serious developmental defect that impact 1 in 10,000 live births. EZH2 has been implicated in numerous human malignancies and is a key therapeutic target in cancer treatment. BRGI and other members ofthe human SWI/SNF family complexes are also strongly implicated in numerous cancers. PROJECT/PERFORIIMNCE SITE(S) (if additional space is needed, use Project/Perfomnance Site Fonnat Page) Pix>ject/Performance Site Primary Location Organizational Name: Massachusetts General Hospital DUNS: 07-313-0411 streeti: 185 Cambridge Street street 2: City: Boston County: state: MA Province: Country: U S A Zip/Postal Code: 0 2 1 1 4

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37GM048405-22
Application #
8515647
Study Section
Special Emphasis Panel (NSS)
Program Officer
Carter, Anthony D
Project Start
1993-01-01
Project End
2018-12-31
Budget Start
2014-01-01
Budget End
2014-12-31
Support Year
22
Fiscal Year
2014
Total Cost
$395,850
Indirect Cost
$168,350
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
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West, Jason A; Cook, April; Alver, Burak H et al. (2014) Nucleosomal occupancy changes locally over key regulatory regions during cell differentiation and reprogramming. Nat Commun 5:4719

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