Natasha Zachara

Image of Dr. Natasha Zachara

Natasha Zachara

Associate Professor
Primary Appointment: 
Biological Chemistry

725. N. Wolfe Street
WBSB 408
Baltimore MD 21205

Research topic: 

O-GlcNAc, A Novel Regulator of the Cellular Stress Response and Cell Surival

The overarching goal of our laboratory is to characterize a novel endogenous protective signaling network of mammalian cells and tissues, in order 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, Cancer). 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, that was discovered at Johns Hopkins by a BCBM graduate student. O-GlcNAc has been implicated in regulating cellular processes as diverse as 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 (mass spectrometry) and genetic manipulation to address these questions.

Selected Publications: 

Zachara NE, O'Donnell N, Mercer JJ, Marth JD, and Hart GW. Dynamic O-GlcNAc modification of nucleocytoplasmic proteins in response to stress. A survival response of mammalian cells. J. Biol. Chem. 2004;279(29):30133-30142.

Jones SP, Zachara NE, Teshima Y, Hart GW, and Marban E. Endogenously-recruitable cardioprotection by N-acetylglucosamine linkage to cellular proteins. Circulation 2008;117(9):1172-82.

Kazemi Z, Chang H, Haserodt SK, McKen C, Zachara NE. O-GlcNAc Regulates Stress-Induced Heat Shock Protein Expression in a GSK-3 Dependent Manner. J. Biol. Chem. 2010;285(50): 39096-39107.

Zachara NE*, Molina H, Wong K, Pandey A, Hart GW. The dynamic stress-induced O-GlcNAcome highlights functions for O-GlcNAc in DNA Repair and other cellular pathways. Amino Acids 2011;40(3):793-808. *Corresponding Author.

Jensen RV, Zachara NE, Nielsen PH, Kimose HH, Kristiansen SB, and Botker HE. Impact of O-GlcNAc on cardioprotection by remote ischemic preconditioning in non-diabetic and diabetic patients. Cardiovascular Research 2013; 97(2):369-378.

Hong J, Martinez M, Sengupta S, Lee A, Wu X, Chaerkady R, O'Meally R, Cole RN, Pandey A, Zachara NE Quantitative Phosphoproteomics Reveals Crosstalk Between Phosphorylation and O-GlcNAc in the DNA Damage Response Pathway. Proteomics 2015;15(2-3):591-607.

Lee A, Henry R, Miller D, Paruchuri VDP, O’Meally R, Boronina T, Cole RN, Zachara NE. Combined Antibody/Lectin-Enrichment Identifies Extensive Changes in the O-GlcNAc Subproteome Upon Oxidative Stress, Journal of Proteome Research; 15(12):4318-4336.

Groves JA, Maduka AO, O’Meally RN, Cole RN, and Zachara NE. Fatty acid synthase: a novel oxidative stress-induced interactor and inhibitor of the O-GlcNAcase. J Biol. Chem. 2017;292(16):6493-6511.

Tan EP, McGreal SR, Graw S, Tessman R, Koppel SJ, Dhakal P, Zhang Z, Machacek M, Zachara NE, Koestler DC, Peterson KR, Thyfault JP, Swerdlow RH, Krishnamurthy P, DiTacchio L, Apte U, Slawson C. Sustained O-GlcNAcylation reprograms mitochondrial function to regulate energy metabolism. J Biol. Chem. 2017; 292(36):14940-14962.

Kim DI, Cutler JA, Na CH, Reckel S, Renuse S, Madugundu AK, Tahir R, Goldschmidt HL, Reddy KL, Huganir RL, Wu X, Zachara NE, Hantschel O, Pandey A. BioSITe: A Method for Direct Detection and Quantitation of Site-Specific Biotinylation. J Proteome Res. 2018;17(2):759-769.

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 128(11):4924-4937. *Co-corresponding authors.

Sager RA, Woodford MR, Backe SJ, Makedon AM, Baker-Williams AJ, Loiselle D, Haystead TA, Zachara NE, Prodromou C, Bourboulia D, Schmidt LS, Linehan WM, Bratslavsky G, Mollapour M. Post-translational Regulation of FNIP1: A rheostat for the molecular chaperons HSP90. Cell Reports, 2019; 26(5):1344-1356.

Umapathi P., Banerjee P.D., Zachara N.E., Abrol N., Wang Q., Mesubi O.O., Luczak E.D., Wu Y., Granger J.M., Wei A-C., Reyes Gaido O.E., Florea L., Talbot Jr. C.C., Hart G.W., Anderson M.E. Excessive O - GlcNAcylation causes heart failure and sudden death. bioRxiv 2020.02.11.943910; doi: (In Revision, Circulation)

Mesubi O.O., Rokita A.G., Abrol N., Wu Y., Chen B., Wang Q., Granger J.M., Luczak E.D., Banerjee P.S., Maier L.S., Wehrens X.H., Pomerantz J.L., Song L-S., Ahima R.S., Zachara N.E., Hart G.W., Anderson M.E. O-GlcNAcylation and oxidation contribute to atrial fibrillation in diabetes by activating CaMKII bioRxiv 2020.02.18.954909; doi: (In Revision, JCI)