Histone methylation plays a fundamental role in the organization of chromatin and in the regulation of gene transcription. Our long-term goal is to elucidate how lysine-specific histone methyltransferases regulate gene expression and contribute to cellular development and disease. One such enzyme that is highly conserved in eukaryotes is the histone H3 lysine 36 (H3K36) methyltransferase Set2. We, and others, have shown that Set2 associates with RNA polymerase II during transcription elongation, and that its methylation at H3K36 directs the recruitment of a histone deacetylase complex (Rpd3S) that suppresses inappropriate initiation of transcription. While the basic functions of Set2 have been characterized, little is still known regarding: i) how the Set2 enzyme itself is regulated, ii) whether other functions for this enzyme exist, and iii) how the distinct methylation states of H3K36 (me1, me2, and me3) and their demethylation contribute to chromatin organization and gene transcription. Using Saccharomyces cerevisiae as a model organism, we plan to use a combination of biochemistry and genetics to further address the functions of Set2 and H3K36me in transcriptional regulation and beyond. Our goal will be to address a number of broad questions that will advance our understanding of how histone methyltransferases and demethylases regulate the chromatin environment and contribute to gene expression. These include: 1) How is Set2 targeted to genes and is itself regulated by post- translational modification? 2) How does H3K36 demethylation contribute to the transcription process? 3) Do the different H3K36 methylation states have distinct biological activities in transcription, and does this histone 'mark'function in other DNA- related activities such as DNA repair and replication? These studies will have a significant impact to the field, as our current understanding of histone post-translational modifications, including H3K36 methylation in particular, is very limited. This is underscored by the fact that the dysregulation of enzymes that mediate H3K36 methylation lead to a variety of human diseases including cancer. Given the complexity of having multiple H3K36-methylating enzymes in mammalian cells, yeast affords the exceptional ability to apply genetics and biochemistry to understand the fundamental functions of a highly significant histone 'mark'in chromatin.

Public Health Relevance

Defects in chromatin organization, DNA packaging and its accessibility is a major cause of human disease, including cancer and numerous developmental defects. Our studies on Set2 will reveal how DNA-based activities such as transcription and repair are regulated, which will address the underlying cause of these public health concerns.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM068088-10
Application #
8197676
Study Section
Molecular Genetics C Study Section (MGC)
Program Officer
Carter, Anthony D
Project Start
2003-05-01
Project End
2013-11-30
Budget Start
2011-12-01
Budget End
2012-11-30
Support Year
10
Fiscal Year
2012
Total Cost
$353,183
Indirect Cost
$111,537
Name
University of North Carolina Chapel Hill
Department
Biochemistry
Type
Schools of Medicine
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Klein, Brianna J; Piao, Lianhua; Xi, Yuanxin et al. (2014) The histone-H3K4-specific demethylase KDM5B binds to its substrate and product through distinct PHD fingers. Cell Rep 6:325-35
Kim, Hyun-Soo; Mukhopadhyay, Rituparna; Rothbart, Scott B et al. (2014) Identification of a BET family bromodomain/casein kinase II/TAF-containing complex as a regulator of mitotic condensin function. Cell Rep 6:892-905
Dronamraju, Raghuvar; Strahl, Brian D (2014) A feed forward circuit comprising Spt6, Ctk1 and PAF regulates Pol II CTD phosphorylation and transcription elongation. Nucleic Acids Res 42:870-81
Klein, Brianna J; Lalonde, Marie-Eve; Côté, Jacques et al. (2014) Crosstalk between epigenetic readers regulates the MOZ/MORF HAT complexes. Epigenetics 9:186-93
Jha, Deepak Kumar; Strahl, Brian D (2014) An RNA polymerase II-coupled function for histone H3K36 methylation in checkpoint activation and DSB repair. Nat Commun 5:3965
Rothbart, Scott B; Strahl, Brian D (2014) Interpreting the language of histone and DNA modifications. Biochim Biophys Acta 1839:627-43
McDaniel, Stephen L; Strahl, Brian D (2013) Stress-free with Rpd3: a unique chromatin complex mediates the response to oxidative stress. Mol Cell Biol 33:3726-7
Gatchalian, Jovylyn; Futterer, Agnes; Rothbart, Scott B et al. (2013) Dido3 PHD modulates cell differentiation and division. Cell Rep 4:148-58
Cai, Ling; Rothbart, Scott B; Lu, Rui et al. (2013) An H3K36 methylation-engaging Tudor motif of polycomb-like proteins mediates PRC2 complex targeting. Mol Cell 49:571-82
Ali, Muzaffar; Rincon-Arano, Hector; Zhao, Wei et al. (2013) Molecular basis for chromatin binding and regulation of MLL5. Proc Natl Acad Sci U S A 110:11296-301

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