This proposal is a competing renewal application of R01 GM62437 that combines the use of chemical approaches, enzymologic analysis, and cellular studies to enhance our understanding of enzymes regulating protein acylation and methylation. It is now well-accepted that post-translational modifications (PTMs) involving lysine acetylation and reversible methylation on histones and other proteins are central to epigenetics. Such epigenetic modifying enzymes are viewed as attractive drug targets for cancer and other diseases. The ghrelin O-acyltransferase (GOAT) enzyme catalyzes the unusual PTM of octanoyl attachment to a Ser side chain of the peptide hormone ghrelin and inhibitors of this enzyme may be useful in the treatment of obesity and diabetes. Although there has been increasing efforts to understand the mechanisms and functions of these PTMs and the enzymes that catalyze them, there are major gaps in our understanding in these areas. Filling these knowledge gaps has the potential to provide a clearer understanding of basic biomedical processes and has the opportunity to enhance the development of novel therapeutic approaches and disease diagnostic strategies. There are four Specific Aims in this proposal. 1. Elucidate the molecular basis for protein substrate selectivity for histone acetyltransferases (HAT) using new chemical and biochemical approaches. We will develop and apply new techniques to generate histone-CoA conjugates and exploit protein microarrays to clarify structural and functional features of HAT-substrate interactions. 2. Develop and apply improved chemical tools for analyzing LSD1 histone demethylation. A combination of synthetic inhibitors and propargyl-histones will be prepared to interrogate LSD1 cellular functions and molecular interactions. 3. Clarify the effects of Lys acetylation on S-adenosyl homocysteine hydrolase and inositol monophosphate dehydrogenase-2. Expressed protein ligation will be used to install acetyl-Lys site-specifically into these metabolic enzymes to dissect potentially important nodes between protein acetylation and cellular metabolism. 4. Define major structural features of ghrelin O-acyltransferase. A combination of membrane topology mapping and bisubstrate analog crosslinking will be used to create a blueprint of this key metabolic regulatory enzyme. We believe that this research effort has the potential to greatly expand our understanding of protein post-translational modification mechanisms and functions and identify new therapeutic opportunities for treating metabolic and neoplastic diseases.

Public Health Relevance

This proposal develops and applies chemical methods for the investigation of protein regulation by post-translational modifications involving acylation and methylation. If successful, it could lead to new therapies for cancer and metabolic diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37GM062437-19
Application #
9314007
Study Section
Special Emphasis Panel (NSS)
Program Officer
Barski, Oleg
Project Start
2001-02-01
Project End
2023-02-28
Budget Start
2018-03-01
Budget End
2019-02-28
Support Year
19
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
Bhat, Shridhar; Hwang, Yousang; Gibson, Matthew D et al. (2018) Hydrazide Mimics for Protein Lysine Acylation To Assess Nucleosome Dynamics and Deubiquitinase Action. J Am Chem Soc 140:9478-9485
Kalin, Jay H; Wu, Muzhou; Gomez, Andrea V et al. (2018) Targeting the CoREST complex with dual histone deacetylase and demethylase inhibitors. Nat Commun 9:53
Weinert, Brian T; Narita, Takeo; Satpathy, Shankha et al. (2018) Time-Resolved Analysis Reveals Rapid Dynamics and Broad Scope of the CBP/p300 Acetylome. Cell 174:231-244.e12
Wu, Mingxuan; Hayward, Dawn; Kalin, Jay H et al. (2018) Lysine-14 acetylation of histone H3 in chromatin confers resistance to the deacetylase and demethylase activities of an epigenetic silencing complex. Elife 7:
Michaelides, Michael R; Kluge, Arthur; Patane, Michael et al. (2018) Discovery of Spiro Oxazolidinediones as Selective, Orally Bioavailable Inhibitors of p300/CBP Histone Acetyltransferases. ACS Med Chem Lett 9:28-33
Cao, Jia; Peng, Jinghua; An, Hongying et al. (2017) Endotoxemia-mediated activation of acetyltransferase P300 impairs insulin signaling in obesity. Nat Commun 8:131
Lasko, Loren M; Jakob, Clarissa G; Edalji, Rohinton P et al. (2017) Discovery of a selective catalytic p300/CBP inhibitor that targets lineage-specific tumours. Nature 550:128-132
Boija, Ann; Mahat, Dig Bijay; Zare, Aman et al. (2017) CBP Regulates Recruitment and Release of Promoter-Proximal RNA Polymerase II. Mol Cell 68:491-503.e5
Zucconi, Beth E; Cole, Philip A (2017) Allosteric regulation of epigenetic modifying enzymes. Curr Opin Chem Biol 39:109-115
Robert, Carine; Nagaria, Pratik K; Pawar, Nisha et al. (2016) Histone deacetylase inhibitors decrease NHEJ both by acetylation of repair factors and trapping of PARP1 at DNA double-strand breaks in chromatin. Leuk Res 45:14-23

Showing the most recent 10 out of 42 publications