The ATP-dependent chromatin remodeler SWI/SNF regulates transcription and DNA recombination bymaking DNA more accessible. The objective of this study is to find the mechanism used by SWI/SNF to moveand disassemble nucleosomes. Nucleosome remodeling will be examined at two levels: changes in histone-DNA interactions and the corresponding changes in the interactions of SWI/SNF with histones and DNA.Ultimately the goal is to correlate these two different sets of interactions with each other in a temporal mannerto provide an unprecedented view of the process of nucleosome remodeling. We propose to apply therelatively new technology of an expanded genetic code to site-specifically incorporate a photoreactive aminoacid analog for examining protein-protein and protein-DNA interactions both in vivo and in vitro. Domains inthe catalytic subunit of SWI/SNF other than the ATPase domain will be systematically studied with the basicpremise that these domains make critical contributions to mobilizing nucleosomes in coordination andcooperation with the ATPase domain. Several domains have already been identified that are necessary forSWI/SNF remodeling. Single molecule magnetic tweezer and DNA unwinding optical trap type experimentswill be done to determine if these domains are important for DNA translocation and nucleosome remodeling.The effect of histone H3 acetylation on SWI/SNF remodeling will be examined to determine at which stage inremodeling it modulates the activity of SWI/SNF and RSC. SWI/SNF has many important regulatory roles such as in stem cell self renewal, cellular differentiation,chromatin maintenance and stability, and oncogenesis. While the yeast system has provided us with importantinsights as to how this enzyme functions and the corresponding models, there still remain many questions asto how SWI/SNF regulates chromatin structure. In this proposal we continue to take advantage of the yeastsystem to examine these questions at a biophysical, biochemical, and molecular genetic level.

Public Health Relevance

.SWI/SNF is involved in regulation of transcription and DNA repair. Often mutations in SWI/SNF areassociated with the onset of cancer and several of the proteins in SWI/SNF are from known tumorsuppressor genes. We are investigating the mechanism of SWI/SNF that is used to regulate theepigenome and how it regulates chromatin structure and function.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
7R01GM048413-20
Application #
8836798
Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Carter, Anthony D
Project Start
1993-01-01
Project End
2016-06-30
Budget Start
2013-12-17
Budget End
2014-06-30
Support Year
20
Fiscal Year
2013
Total Cost
$146,907
Indirect Cost
$55,090
Name
University of Texas MD Anderson Cancer Center
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
800772139
City
Houston
State
TX
Country
United States
Zip Code
77030
Bartholomew, Blaine (2014) Regulating the chromatin landscape: structural and mechanistic perspectives. Annu Rev Biochem 83:671-96
Hota, Swetansu K; Bartholomew, Blaine (2011) Diversity of operation in ATP-dependent chromatin remodelers. Biochim Biophys Acta 1809:476-87
Gangaraju, Vamsi K; Bartholomew, Blaine (2007) Mechanisms of ATP dependent chromatin remodeling. Mutat Res 618:3-17