Natasha Zachara
725. N. Wolfe Street
WBSB 408
Baltimore MD 21205
The overarching goal of our laboratory is to characterize a novel endogenous protective signaling network of mammalian cells and tissues to provide insight into the molecular mechanisms underlying disease and to highlight novel therapeutic avenues.
Cells and tissues respond to environmental and physiological injury by reprogramming transcription, translation, metabolism, and signal transduction to affect repair and survival, and if necessary to promote programmed cell death. Collectively, this cell-wide reprogramming is known as the cellular stress response and is characterized by the induction of chaperones known as heat shock proteins (HSP). The cellular stress response plays a critical role in the response of tissues to a broad range of physiological insults (stroke, heart attack, DNA damage) and dysregulation of this response is implicated in the pathogenesis of a host of diseases (proteinopathies such as Alzheimer’s disease). Our laboratory has demonstrated that the glycan modification, O-GlcNAc, is a critical regulator of the cellular stress response that is relevant to numerous clinical models.
O-GlcNAc is a glycan modification of thousands of intracellular proteins such as transcription factors, kinases, and cytoskeletal proteins. O-GlcNAc has been implicated in regulating cellular processes including protein folding, localization, degradation, activity, post-translational modifications, and interactions. The cell co-ordinates these molecular events, on thousands of cellular proteins, in concert with environmental and physiological cues, to fine-tune epigenetics, transcription, translation, signal transduction, cell cycle, and metabolism. The cellular stress response is no exception: diverse forms of injury result in dynamic changes to the O-GlcNAc subproteome that promote survival. Research in the laboratory is broadly focused on answering two questions in cell models of oxidative stress and ex vivo models of cardiac ischemia reperfusion injury: Firstly, which proteins are dynamically O-GlcNAc modified in response to injury and how does this simple sugar modify protein function to promote cell survival. Secondly, how are the enzymes that add and remove O-GlcNAc regulated at times of injury. We utilize traditional biochemical and molecular biology based techniques in combination with high throughput technologies and genetic manipulation to address these questions.
Martinez M, Renuse S, Kreimer S, O’Meally R, Natov P, Madugundu AK, Nirujogi RS, Tahir R, Cole R, Pandey A, Zachara NE. Quantitative Proteomics Reveals that the OGT Interactome Is Remodeled in Response to Oxidative Stress. Mol Cell Proteomics. 2021 Mar 12;20:100069. doi: 10.1016/j.mcpro.2021.100069. Epub ahead of print. PMID: 33716169; PMCID: PMC8079276.
Umapathi P, Mesubi OO, Banerjee PS, Abrol N, Wang Q, Luczak ED, Wu Y, Granger JM, Wei AC, Reyes Gaido OE, Florea L, Talbot CC Jr, Hart GW, Zachara NE, Anderson ME. Excessive O-GlcNAcylation Causes Heart Failure and Sudden Death. Circulation. 2021 Apr 27;143(17):1687-1703. doi: 10.1161/CIRCULATIONAHA.120.051911. Epub 2021 Feb 17. Erratum in: Circulation. 2021 Apr 27;143(17):e892. PMID: 33593071; PMCID: PMC8085112.
Mesubi OO, Rokita AG, Abrol N, Wu Y, Chen B, Wang Q, Granger JM, Tucker-Bartley A, Luczak ED, Murphy KR, Umapathi P, Banerjee PS, Boronina TN, Cole RN, Maier LS, Wehrens XH, Pomerantz JL, Song LS, Ahima RS, Hart GW, Zachara NE, Anderson ME. Oxidized CaMKII and O-GlcNAcylation cause increased atrial fibrillation in diabetic mice by distinct mechanisms. J Clin Invest. 2021 Jan 19;131(2):e95747. doi: 10.1172/JCI95747. PMID: 33151911; PMCID: PMC7810480.
Taparra K, Wang H, Malek R, Lafargue A, Barbhuiya MA, Wang X, Simons BW, Ballew M, Nugent K, Groves J, Williams RD, Shiraishi T, Verdone J, Yildirir G, Henry R, Zhang B, Wong J, Wang KK, Nelkin BD, Pienta KJ, Felsher D, Zachara NE*, Tran PT*. O-GlcNAcylation is required for mutant KRAS-induced lung tumorigenesis. J Clin Invest. 2018 Nov 1;128(11):4924-4937. doi: 10.1172/JCI94844. Epub 2018 Sep 24. PMID: 30130254; PMCID: PMC6205381.*Co-corresponding authors.
Groves JA, Maduka AO, O’Meally RN, Cole RN, Zachara NE. Fatty acid synthase inhibits the O-GlcNAcase during oxidative stress. J Biol Chem. 2017 Apr 21;292(16):6493-6511. doi: 10.1074/jbc.M116.760785. Epub 2017 Feb 23. PMID: 28232487; PMCID: PMC5399103.
Lee A, Miller D, Henry R, Paruchuri VD, O’Meally RN, Boronina T, Cole RN, Zachara NE. Combined Antibody/Lectin Enrichment Identifies Extensive Changes in the O-GlcNAc Sub-proteome upon Oxidative Stress. J Proteome Res. 2016 Dec 2;15(12):4318-4336. doi: 10.1021/acs.jproteome.6b00369. Epub 2016 Oct 14. PMID: 27669760.
Zhong J, Martinez M, Sengupta S, Lee A, Wu X, Chaerkady R, Chatterjee A, O’Meally RN, Cole RN, Pandey A, Zachara NE. Quantitative phosphoproteomics reveals crosstalk between phosphorylation and O-GlcNAc in the DNA damage response pathway. Proteomics. 2015 Jan;15(2-3):591-607. doi: 10.1002/pmic.201400339. PMID: 25263469; PMCID: PMC4564869.
Kazemi Z, Chang H, Haserodt S, McKen C, Zachara NE. O-linked beta-N-acetylglucosamine (O-GlcNAc) regulates stress-induced heat shock protein expression in a GSK-3beta-dependent manner. J Biol Chem. 2010 Dec 10;285(50):39096-107. doi: 10.1074/jbc.M110.131102. Epub 2010 Oct 6. PMID: 20926391; PMCID: PMC2998145.
Zachara NE*, Molina H, Wong KY, Pandey A, Hart GW. The dynamic stress-induced “O-GlcNAc-ome” highlights functions for O-GlcNAc in regulating DNA damage/repair and other cellular pathways. Amino Acids. 2011 Mar;40(3):793-808. doi: 10.1007/s00726-010-0695-z. Epub 2010 Jul 31. PMID: 20676906; PMCID: PMC3329784. *Corresponding Author
Jones SP, Zachara NE, Ngoh GA, Hill BG, Teshima Y, Bhatnagar A, Hart GW, Marbán E. Cardioprotection by N-acetylglucosamine linkage to cellular proteins. Circulation. 2008 Mar 4;117(9):1172-82. doi: 10.1161/CIRCULATIONAHA.107.730515. Epub 2008 Feb 19. PMID: 18285568.
Zachara NE, O’Donnell N, Cheung WD, Mercer JJ, Marth JD, Hart GW. Dynamic O-GlcNAc modification of nucleocytoplasmic proteins in response to stress. A survival response of mammalian cells. J Biol Chem. 2004 Jul 16;279(29):30133-42. doi: 10.1074/jbc.M403773200. Epub 2004 May 11. PMID: 15138254.