The Mixed Lineage Leukemia protein-1 (MLL1) catalyzes histone H3 lysine 4 (H3K4) methylation, which is an epigenetic mark essential for the regulation of HOX genes in hematopoiesis and development. Translocations that disrupt the MLL1 gene are present in a unique group of acute leukemias, often predicting a poor prognosis. Other MLL1 rearrangements and amplifications increase MLL1's enzymatic activity and are oncogenic. MLL1 contains an evolutionarily conserved ~130 amino acid SET domain that catalyzes H3K4 methylation. Recent studies indicate that the enzymatic activity of MLL1 is regulated by a conserved complex of proteins including WDR5, RbBP5, and ASH2L. These proteins form an independent complex that binds to MLL1 and regulates its ability to mono-, di-, or trimethylate H3K4 substrates, a phenomenon known as 'Product Specificity'. Since different levels of methylation of H3K4 are associated with different transcriptional outcomes, it is imperative to understand the molecular mechanisms by which the product specificity of MLL1 is regulated. Despite the important biological role of MLL1 and its involvement in human leukemia, there is currently little information about the protein-structural features that are responsible for the enzymatic activity of the MLL1 core complex. The long-term goal of this research is to fully characterize the mechanisms for the regulation of H3K4 methylation by the MLL1 core complex. This proposal takes a structure-function approach to investigate the molecular mechanisms for the enzymatic activity of the MLL1 core complex. We have obtained novel preliminary results indicating that the intrinsic product specificity of the MLL1 SET domain is that of a slow monomethyltransferase, but becomes a fast dimethyltransferase when in complex with WDR5, RbBP5, and Ash2L. Here we propose to determine the mechanisms responsible for H3K4 dimethylation activity of the MLL1 core complex. In addition, we will determine the protein-structural features that account for its assembly. To address these aims, we combine molecular, biochemical, and biophysical approaches to investigate the regulation of H3K4 methylation by the MLL1 core complex. This information will increase our understanding of a key enzyme complex and how it is regulated in the pathways that control transcriptional activation in eukaryotes. This investigation is important because it may lead to better diagnostics and the rational design of anti-cancer drugs that inhibit MLL1's enzymatic activity.

Public Health Relevance

The mechanisms that regulate gene transcription in eukaryotes are not well understood. We will employ a biophysical analysis of the Mixed Lineage Leukemia protein-1 (MLL1) core complex to gain insight into its role in transcriptional regulation. We expect this knowledge will lead to the development of anti-cancer agents for the treatment of certain human leukemias.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
7R01CA140522-03
Application #
8206678
Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Howcroft, Thomas K
Project Start
2010-01-01
Project End
2014-12-31
Budget Start
2012-05-31
Budget End
2012-12-31
Support Year
3
Fiscal Year
2012
Total Cost
$289,161
Indirect Cost
$87,886
Name
Upstate Medical University
Department
Surgery
Type
Schools of Medicine
DUNS #
058889106
City
Syracuse
State
NY
Country
United States
Zip Code
13210
Alicea-Velázquez, Nilda L; Shinsky, Stephen A; Loh, Daniel M et al. (2016) Targeted Disruption of the Interaction between WD-40 Repeat Protein 5 (WDR5) and Mixed Lineage Leukemia (MLL)/SET1 Family Proteins Specifically Inhibits MLL1 and SETd1A Methyltransferase Complexes. J Biol Chem 291:22357-22372
Shinsky, Stephen A; Monteith, Kelsey E; Viggiano, Susan et al. (2015) Biochemical reconstitution and phylogenetic comparison of human SET1 family core complexes involved in histone methylation. J Biol Chem 290:6361-75
Shinsky, Stephen A; Cosgrove, Michael S (2015) Unique Role of the WD-40 Repeat Protein 5 (WDR5) Subunit within the Mixed Lineage Leukemia 3 (MLL3) Histone Methyltransferase Complex. J Biol Chem 290:25819-33
Tie, Feng; Banerjee, Rakhee; Saiakhova, Alina R et al. (2014) Trithorax monomethylates histone H3K4 and interacts directly with CBP to promote H3K27 acetylation and antagonize Polycomb silencing. Development 141:1129-39
Patel, Anamika; Vought, Valarie E; Swatkoski, Stephen et al. (2014) Automethylation activities within the mixed lineage leukemia-1 (MLL1) core complex reveal evidence supporting a "two-active site" model for multiple histone H3 lysine 4 methylation. J Biol Chem 289:868-84
Shinsky, Stephen A; Hu, Michael; Vought, Valarie E et al. (2014) A non-active-site SET domain surface crucial for the interaction of MLL1 and the RbBP5/Ash2L heterodimer within MLL family core complexes. J Mol Biol 426:2283-99
Zhong, Shurong; Hsu, FoSheng; Stefan, Christopher J et al. (2012) Allosteric activation of the phosphoinositide phosphatase Sac1 by anionic phospholipids. Biochemistry 51:3170-7
Cosgrove, Michael S (2012) Writers and readers: deconvoluting the harmonic complexity of the histone code. Nat Struct Mol Biol 19:739-40
Cosgrove, Michael S; Ding, Ye; Rennie, William A et al. (2012) The Bin3 RNA methyltransferase targets 7SK RNA to control transcription and translation. Wiley Interdiscip Rev RNA 3:633-47
Dharmarajan, Venkatasubramanian; Lee, Jeong-Heon; Patel, Anamika et al. (2012) Structural basis for WDR5 interaction (Win) motif recognition in human SET1 family histone methyltransferases. J Biol Chem 287:27275-89

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