The overall goal of this proposal is to understand how ATP-dependent chromatin remodelers influence diverse biological processes. The experiments outlined here focus on understanding how the CSB (Cockayne Syndrome complementation group B) chromatin remodeler modulates chromatin structure and how defects in this protein lead to disease. ATPdependent chromatin remodelers alter chromatin structure non-covalently to regulate DNA access and/or space nucleosomes. Thus, these activities play important roles in regulating nuclear processes such as transcription, replication and DNA repair as well as developmental events including transcriptional memory and cell-fate determination. Mutations in CSB lead to Cockayne syndrome: a disease in which patients are highly sensitive to UV light, age prematurely, and suffer from profound developmental and neurological disorders. CSB plays a pivotal role in transcription-coupled DNA repair;however, little is known as to how its remodeling activity influences this process. The availability of mutant CSB cell lines that phenocopy some of the defects associated with Cockayne syndrome (such as UV sensitivity) makes CSB an excellent candidate remodeler to elucidate the relationship between the biochemical activities and biological functions of ATPdependent chromatin remodeling complexes. To obtain greater insight into the mechanism whereby CSB remodels chromatin, and to elucidate the contribution of this activity to normal development, DNA repair and disease, we propose three aims (1) Dissect the mechanism of CSB remodeling activity, (2) Dissect the functions of CSB in transcription-coupled DNA repair, and (3) Investigate the impact of CSB-binding proteins on CSB function. PUBLIC HEALTH REVELANCE: Chromatin is the fundamental template for all nuclear process and there are countless examples of human disease resulting from misregulation of chromatin structure, including cancer. This proposed research project will not only reveal the underlying mechanism that directly lead to the devastating disease Cockayne syndrome, but offer important insights into the fundamental principles that regulate chromatin structure.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM084983-06
Application #
8304965
Study Section
Nuclear Dynamics and Transport (NDT)
Program Officer
Janes, Daniel E
Project Start
2008-08-01
Project End
2014-07-31
Budget Start
2012-08-01
Budget End
2014-07-31
Support Year
6
Fiscal Year
2012
Total Cost
$297,950
Indirect Cost
$111,731
Name
University of Pennsylvania
Department
Biochemistry
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Lake, Robert J; Boetefuer, Erica L; Won, Kyoung-Jae et al. (2016) The CSB chromatin remodeler and CTCF architectural protein cooperate in response to oxidative stress. Nucleic Acids Res 44:2125-35
Arora, Sanjeevani; Yan, Hong; Cho, Iltaeg et al. (2015) Genetic Variants That Predispose to DNA Double-Strand Breaks in Lymphocytes From a Subset of Patients With Familial Colorectal Carcinomas. Gastroenterology 149:1872-1883.e9
Lake, Robert J; Boetefuer, Erica L; Tsai, Pei-Fang et al. (2014) The sequence-specific transcription factor c-Jun targets Cockayne syndrome protein B to regulate transcription and chromatin structure. PLoS Genet 10:e1004284
Lake, Robert J; Tsai, Pei-Fang; Choi, Inchan et al. (2014) RBPJ, the major transcriptional effector of Notch signaling, remains associated with chromatin throughout mitosis, suggesting a role in mitotic bookmarking. PLoS Genet 10:e1004204
Lake, Robert J; Fan, Hua-Ying (2013) Structure, function and regulation of CSB: a multi-talented gymnast. Mech Ageing Dev 134:202-11
Cho, Iltaeg; Tsai, Pei-Fang; Lake, Robert J et al. (2013) ATP-dependent chromatin remodeling by Cockayne syndrome protein B and NAP1-like histone chaperones is required for efficient transcription-coupled DNA repair. PLoS Genet 9:e1003407
Lake, Robert J; Basheer, Asjad; Fan, Hua-Ying (2011) Reciprocally regulated chromatin association of Cockayne syndrome protein B and p53 protein. J Biol Chem 286:34951-8
Kelly, Deborah F; Lake, Robert J; Middelkoop, Teije C et al. (2010) Molecular structure and dimeric organization of the Notch extracellular domain as revealed by electron microscopy. PLoS One 5:e10532
Lake, Robert J; Geyko, Anastasia; Hemashettar, Girish et al. (2010) UV-induced association of the CSB remodeling protein with chromatin requires ATP-dependent relief of N-terminal autorepression. Mol Cell 37:235-46