The PHD finger (Plant Homeodomain) module is a signature chromatin-associated protein motif. This module is present throughout eukaryotic proteomes, and mutations in the PHD fingers of many proteins are associated with cancers, immunodeficiency and mental retardation syndromes, and other genetic disorders. We previously demonstrated that a subset of PHD fingers act as high affinity binding modules for histone H3 trimethylated at lysine 4 (H3K4me3). We linked H3K4me3 to multiple different functions via its recognition by discrete PHD finger nuclear proteins, including providing the first evidence that disrupting the read-out of a histone modification can cause an inherited human disease. Our long-term goal is to develop a comprehensive understanding of how PHD domain-containing proteins impact on chromatin dynamics and the relationship of such activities to fundamental nuclear functions and human disease processes. Here we focus on the biology and function of the multiple PHD domain-containing proteins NSD2 (also named MMSET and WHSC1) and NSD3 (also named WHSC1L), two histone lysine methyltransferases implicated in cancer pathogenesis. However, the molecular mechanisms by which these enzymes regulate chromatin and the relationship of their enzymatic activities to disease pathogenesis in vitro and in vivo is not well understood. We hypothesize that NSD2 and NSD3, via regulation of H3K36 methylation dynamics, govern nuclear and epigenetic programs important for normal and oncogenic cellular behaviors.
In Aim 1, we characterize the molecular mode of action of H3K36. We continue our studies of H3K36me- binding proteins and through further development refine a proteomic platform for discovery of new proteins that preferentially recognize H3K36me2. We test the hypothesis that these proteins transduce NSD2 activity at chromatin to downstream biological outcomes. The goal of Aim 2 is to explore the mode of action of NSD2 in cancer in vivo, which has not previously been done. We will use a novel conditional NSD2 overexpression allele to test the hypothesis that in a wild-type microenvironment increased NSD2 expression and enzymatic activity reprograms the epigenome to stimulate cancer development.
In Aim 3, we focus on elucidating the physiologic catalytic activity of NSD3 and its relationship to chromatin and disease regulation. We will use a combination of strategies and approaches to help characterize the mechanisms underlying NSD3 activity in normal and cancer cells. Together these studies will provide important new insights into how PHD finger proteins and histone methylation dynamics regulates fundamental nuclear processes and the relationship of these activities to the pathogenesis of human diseases.

Public Health Relevance

We propose to investigate the molecular mode of action of two enzymes that regulate a key epigenetic modification on histone proteins in mammalian cells. Numerous human diseases, including cancer arise from epigenetic abnormalities. This proposal will provide new insight into how epigenetic mechanisms regulate important cellular functions, and has the potential to identify new targets for therapeutic intervention for human disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM079641-11
Application #
9470897
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Carter, Anthony D
Project Start
2007-03-01
Project End
2021-02-28
Budget Start
2018-03-01
Budget End
2019-02-28
Support Year
11
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Stanford University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94304
Sankaran, Saumya M; Gozani, Or (2017) Characterization of H3.3K36M as a tool to study H3K36 methylation in cancer cells. Epigenetics 12:917-922
Zhu, Li; Li, Qin; Wong, Stephen H K et al. (2016) ASH1L Links Histone H3 Lysine 36 Dimethylation to MLL Leukemia. Cancer Discov 6:770-83
Li, Sisi; Yang, Zhenlin; Du, Xuan et al. (2016) Structural Basis for the Unique Multivalent Readout of Unmodified H3 Tail by Arabidopsis ORC1b BAH-PHD Cassette. Structure 24:486-94
Huang, Wei-Hsiang; Guenthner, Casey J; Xu, Jin et al. (2016) Molecular and Neural Functions of Rai1, the Causal Gene for Smith-Magenis Syndrome. Neuron 92:392-406
van Nuland, Rick; Gozani, Or (2016) Histone H4 Lysine 20 (H4K20) Methylation, Expanding the Signaling Potential of the Proteome One Methyl Moiety at a Time. Mol Cell Proteomics 15:755-64
Sankaran, Saumya M; Wilkinson, Alex W; Elias, Joshua E et al. (2016) A PWWP Domain of Histone-Lysine N-Methyltransferase NSD2 Binds to Dimethylated Lys-36 of Histone H3 and Regulates NSD2 Function at Chromatin. J Biol Chem 291:8465-74
Chen, Shoudeng; Yang, Ze; Wilkinson, Alex W et al. (2015) The PZP Domain of AF10 Senses Unmodified H3K27 to Regulate DOT1L-Mediated Methylation of H3K79. Mol Cell 60:319-27
Zhang, Wei; Sankaran, Saumya; Gozani, Or et al. (2015) A Meier-Gorlin syndrome mutation impairs the ORC1-nucleosome association. ACS Chem Biol 10:1176-80
Carlson, Scott M; Moore, Kaitlyn E; Sankaran, Saumya M et al. (2015) A Proteomic Strategy Identifies Lysine Methylation of Splicing Factor snRNP70 by the SETMAR Enzyme. J Biol Chem 290:12040-7
Moore, Kaitlyn E; Gozani, Or (2014) An unexpected journey: lysine methylation across the proteome. Biochim Biophys Acta 1839:1395-403

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