Nucleosomes and other components of chromatin can repress transcription by blocking the access of transcription factors and other regulatory proteins to DNA. Interactions between nucleosomes and linker histones promote the formation of 30 nm fibers and increasingly compact forms of higher-order chromatin structure, further limiting the access of regulatory proteins to DNA. Alterations in chromatin structure lead to a variety of human diseases, including cancer and birth defects. Although tremendous progress has been made toward determining the mechanism of action of chromatin-remodeling factors and histone-modifying enzymes, much remains to be learned about how they regulate transcription by altering the structure and spacing of nucleosomes. Even less is known about how higher-order chromatin structure is regulated and used to control gene expression in eukaryotic cells. To address these important issues, our laboratory uses Drosophila melanogaster as a model organism to study the function of chromatin-remodeling and modifying enzymes. Much of our work has been focused on the roles of Polycomb and trithorax group in transcription and development. Polycomb group proteins play important roles in the regulation of cellular pluripotency and differentiation by methylating lysine 27 of histone H3 over broad chromatin domains. This chromatin modification plays a key role in epigenetic gene silencing in organisms ranging from flies to humans. A key issue concerns how Polycomb group repression is overcome to permit the expression of genes required for differentiation. Recent studies in our lab have suggested that Kismet (KIS), an ATP-dependent chromatin-remodeling factor, plays an important role in this process by preventing the spread of H3K27 methylation into active genes. To test this hypothesis, we will compare the genome-wide distributions of KIS and H3K27 methylation and determine if KIS acts as a barrier to the spread of the repressive modification. Other studies in our laboratory have suggested that another chromatin-remodeling factor, Imitation-SWI (ISWI), plays a global role in chromatin compaction and transcriptional repression by promoting the association of the linker histone H1 with chromatin. To test this hypothesis, we will compare changes in gene expression resulting from the loss of ISWI and histone H1 function in vivo. We will also use complementary genetic, biochemical and cell biological approaches to determine how ISWI promotes the association of histone H1 with chromatin and identify other factors involved in this process. By studying multiple chromatin- remodeling factors in a single model organism, we will gain a much better understanding of their roles in transcription, development and disease.

Public Health Relevance

This goal of this project is to understand how Drosophila chromatin-remodeling factors, including KIS-L and ISWI, control gene expression and development by altering chromatin structure. Mutations in the human counterparts of these proteins are responsible for numerous cancers and birth defects, including CHARGE syndrome, a serious developmental disorder affecting 1 in 8,000 births. The information gained from this project will therefore be directly relevant to human health.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM049883-20
Application #
8429511
Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Carter, Anthony D
Project Start
1993-08-01
Project End
2015-02-28
Budget Start
2013-03-01
Budget End
2015-02-28
Support Year
20
Fiscal Year
2013
Total Cost
$345,339
Indirect Cost
$117,392
Name
University of California Santa Cruz
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
125084723
City
Santa Cruz
State
CA
Country
United States
Zip Code
95064
Dorighi, Kristel M; Tamkun, John W (2013) The trithorax group proteins Kismet and ASH1 promote H3K36 dimethylation to counteract Polycomb group repression in Drosophila. Development 140:4182-92
Siriaco, Giorgia; Deuring, Renate; Chioda, Mariacristina et al. (2009) Drosophila ISWI regulates the association of histone H1 with interphase chromosomes in vivo. Genetics 182:661-9
Srinivasan, Shrividhya; Dorighi, Kristel M; Tamkun, John W (2008) Drosophila Kismet regulates histone H3 lysine 27 methylation and early elongation by RNA polymerase II. PLoS Genet 4:e1000217
Burgio, Giosalba; La Rocca, Gaspare; Sala, Anna et al. (2008) Genetic identification of a network of factors that functionally interact with the nucleosome remodeling ATPase ISWI. PLoS Genet 4:e1000089
Corona, Davide F V; Siriaco, Giorgia; Armstrong, Jennifer A et al. (2007) ISWI regulates higher-order chromatin structure and histone H1 assembly in vivo. PLoS Biol 5:e232
Srinivasan, Shrividhya; Armstrong, Jennifer A; Deuring, Renate et al. (2005) The Drosophila trithorax group protein Kismet facilitates an early step in transcriptional elongation by RNA Polymerase II. Development 132:1623-35
Armstrong, Jennifer A; Sperling, Adam S; Deuring, Renate et al. (2005) Genetic screens for enhancers of brahma reveal functional interactions between the BRM chromatin-remodeling complex and the delta-notch signal transduction pathway in Drosophila. Genetics 170:1761-74
Silva, Elizabeth; Tiong, Stanley; Pedersen, Michael et al. (2004) ATM is required for telomere maintenance and chromosome stability during Drosophila development. Curr Biol 14:1341-7
Corona, Davide F; Armstrong, Jennifer A; Tamkun, John W (2004) Genetic and cytological analysis of Drosophila chromatin-remodeling factors. Methods Enzymol 377:70-85
Corona, Davide F V; Tamkun, John W (2004) Multiple roles for ISWI in transcription, chromosome organization and DNA replication. Biochim Biophys Acta 1677:113-9

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