The overall goal of research in the Watt lab is tounderstand the biological roles of lysine deacetylases (KDACs) and thebiochemical mechanisms underlying those functions.
Metal-dependent lysine deacetylases (KDACs, also known ashistone deacetylases or HDACs) are multi-functional proteins that mediatecontrol of numerous cellular processes and have been implicated in manycancers. KDACs regulate the behavior ofother cellular proteins through control of the post-translational modificationof lysine residues. Acetylation anddeacetylation of proteins have been directly associated with a wide range ofbiological processes, including metabolic regulation and organismal developmentas well as numerous cancers and other diseases. Using a combination of in vitro and computational techniques, we areinvestigating the specific molecular features of KDACs leading to non-histonesubstrate selection and catalytic activity. Mapping the interactions in each KDAC active site that determineselectivity provides new design strategies for more targeted inhibitors asdrugs leads.
Using gene-edited cell lines that selectively control KDACcatalytic activity while retaining endogenous expression, we are alsoinvestigating the specific non-histone targets of several KDACs, the cellularcomponents that regulate KDAC activity, and the functional roles of KDACs. Improved understanding of the specificinteractions and substrates of KDACs will suggest novel targets for cancertreatment. Furthermore, there is limitedunderstanding of the function of acetylation itself on proteins undergoing themodification, in part because the factors regulating the addition and removalof the modification are not well-characterized. Enhanced understanding of the core biology associated with KDACs hasdirect relevance to understanding the impact of increased or decreased KDACexpression that has been reported in numerous cancers, and hence when targetingthe KDAC or a downstream target of a KDAC may be an appropriate therapeuticstrategy.
Key Words
Histone Deacetylase (HDAC)
Lysine Deacetylase (KDAC)
Gene Ontology
Gene Editing
RNA-seq
Substrate Specificity
Post-translational Modification (PTM)
Enzyme Kinetics
Molecular Dynamics
Education
Select Publications
Toro TB, Edenfield SA, Hylton BJ, Watt TJ. Chelatable trace zinc causes low, irreproducible KDAC8 activity. Anal. Biochem. 2018, 540-541, 9-14. PMID: 29100752
Toro TB, Painter RG, Haynes RA, Glotser EY, Bratton MR, Bryant JR, Nichols KA, Matthew-Onabanjo AN, Matthew AN, Bratcher DR, Perry CD, Watt TJ. Purification of metal-dependent lysine deacetylases with consistently high activity. Protein Exp. Purif. 2018, 141, 1-6. PMID: 28843507
Toro TB, Bryant JR, Watt TJ. Lysine deacetylases exhibit distinct changes in activity profiles due to fluorophore-conjugation of substrates. Biochemistry 2017, 56, 4549-4558. PMID: 28749131
Toro TB, Pingali S, Nguyen TP, Garrett DS, Dodson KA, Nichols KA, Haynes RA, Payton-Stewart F, Watt TJ. KDAC8 with high basal velocity is not activated by N-acetylthioureas. PLoS One 2016, 11, e0146900. PMID: 26745872
Toro TB, Watt TJ. KDAC8 substrate specificity quantified by a biologically-relevant, label-free deacetylation assay. Protein Sci. 2015, 24, 2020-2032. PMID: 26402585
Johanson KE, Watt TJ. Inquiry-based experiments for large-scale introduction to PCR and restriction enzyme digests. Biochem. Mol. Bio. Ed. 2015, 43, 441-448. PMID: 26503481
Toro TB, Nguyen TP, Watt TJ. An improved 96-well turbidity assay for T4 lysozyme activity. MethodsX 2015, 2, 256-262. PMID: 26150996
Johanson KE, Watt TJ, McIntyre NR, Thompson M. Purification and characterization of enzymes from yeast: an extended undergraduate laboratory sequence for large classes. Biochem. Mol. Bio. Ed. 2013, 41, 251-261. PMID: 23868379
