Skip to main content
SpringerLink
Log in

The dynamic stress-induced “O-GlcNAc-ome” highlights functions for O-GlcNAc in regulating DNA damage/repair and other cellular pathways

  • Original Article
  • Published:
Amino Acids Aims and scope Submit manuscript

Abstract

The modification of nuclear, mitochondrial, and cytoplasmic proteins by O-linked β-N-acetylglucosamine (O-GlcNAc) is a dynamic and essential post-translational modification of metazoans. Numerous forms of cellular injury lead to elevated levels of O-GlcNAc in both in vivo and in vitro models, and elevation of O-GlcNAc levels before, or immediately after, the induction of cellular injury is protective in models of heat stress, oxidative stress, endoplasmic reticulum (ER) stress, hypoxia, ischemia reperfusion injury, and trauma hemorrhage. Together, these data suggest that O-GlcNAc is a regulator of the cellular stress response. However, the molecular mechanism(s) by which O-GlcNAc regulates protein function leading to enhanced cell survival have not been identified. In order to determine how O-GlcNAc modulates stress tolerance in these models we have used stable isotope labeling with amino acids in cell culture to determine the identity of proteins that undergo O-GlcNAcylation in response to heat shock. Numerous proteins with diverse functions were identified, including NF-90, RuvB-like 1 (Tip49α), RuvB-like 2 (Tip49β), and several COPII vesicle transport proteins. Many of these proteins bind double-stranded DNA-dependent protein kinase (PK), or double-stranded DNA breaks, suggesting a role for O-GlcNAc in regulating DNA damage signaling or repair. Supporting this hypothesis, we have shown that DNA-PK is O-GlcNAc modified in response to numerous forms of cellular stress.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

Carm1 CK II:

Casein kinase II

DNA-PK:

DNA-dependent protein kinase

DMSO:

Dimethylsulfoxide

DON:

6-diazo-5-oxonorleucine

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

FBS:

Fetal bovine serum

HSP:

Heat shock protein

MEFs:

Mouse embryonic fibroblasts

O-GlcNAc:

Monosaccharides of O-linked β-N-acetylglucosamine

O-GlcNAcase:

O-GlcNAc hexosaminidase (EC 3.2.1.52)

OGT: UDP-GlcNAc:

Polypeptide O-β- N-acetylglucosaminyltransferase (EC 2.4.1.94)

PBS:

Phosphate-buffered saline

PUGNAc:

O-(2-acetamido-2-deoxy-d-glucopyranosylidene) amino-N-phenylcarbamate

TBS:

Tris-buffered saline

References

  • Amanchy R, Kalume DE, Pandey A (2005) Stable isotope labeling with amino acids in cell culture (SILAC) for studying dynamics of protein abundance and posttranslational modifications. Sci STKE 2005, p l2

  • Amodio G, Renna M, Paladino S, Venturi C, Tacchetti C, Moltedo O, Franceschelli S, Mallardo M, Bonatti S, Remondelli P (2009) Endoplasmic reticulum stress reduces the export from the ER and alters the architecture of post-ER compartments. Int J Biochem Cell Biol 41:2511–2521

    Article  CAS  PubMed  Google Scholar 

  • Barr FD, Krohmer LJ, Hamilton JW, Sheldon LA (2009) Disruption of histone modification and CARM1 recruitment by arsenic represses transcription at glucocorticoid receptor-regulated promoters. PLoS One 4:e6766

    Article  PubMed  Google Scholar 

  • Bauer I, Graessle S, Loidl P, Hohenstein K, Brosch G (2010) Novel insights into the functional role of three protein arginine methyltransferases in Aspergillus nidulans. Fungal Genet Biol

  • Belles-Boix E, Babiychuk E, Montagu MV, Inze D, Kushnir S (2000) CEF, a sec24 homologue of Arabidopsis thaliana, enhances the survival of yeast under oxidative stress conditions. J Exp Bot 51:1761–1762

    Article  CAS  PubMed  Google Scholar 

  • Champattanachai V, Marchase RB, Chatham JC (2007) Glucosamine protects neonatal cardiomyocytes from ischemia-reperfusion injury via increased protein-associated O-GlcNAc. Am J Physiol Cell Physiol 292:C178–C187

    Article  CAS  PubMed  Google Scholar 

  • Champattanachai V, Marchase RB, Chatham JC (2008) Glucosamine protects neonatal cardiomyocytes from ischemia-reperfusion injury via increased protein O-GlcNAc and increased mitochondrial Bcl-2. Am J Physiol Cell Physiol 294:C1509–C1520

    Article  CAS  PubMed  Google Scholar 

  • Chatham JC, Not LG, Fulop N, Marchase RB (2008) Hexosamine biosynthesis and protein O-glycosylation: the first line of defense against stress, ischemia, and trauma. Shock 29:431–440

    CAS  PubMed  Google Scholar 

  • Chen SL, Loffler KA, Chen D, Stallcup MR, Muscat GE (2002) The coactivator-associated arginine methyltransferase is necessary for muscle differentiation: CARM1 coactivates myocyte enhancer factor-2. J Biol Chem 277:4324–4333

    Article  CAS  PubMed  Google Scholar 

  • Cheng D, Cote J, Shaaban S, Bedford MT (2007) The arginine methyltransferase CARM1 regulates the coupling of transcription and mRNA processing. Mol Cell 25:71–83

    Article  PubMed  Google Scholar 

  • Cheng Y, Liu X, Zhang S, Lin Y, Yang J, Zhang C (2009) MicroRNA-21 protects against the H(2)O(2)-induced injury on cardiac myocytes via its target gene PDCD4. J Mol Cell Cardiol 47:5–14

    Article  CAS  PubMed  Google Scholar 

  • Cheung WD, Hart GW (2008) AMP-activated protein kinase and p38 MAPK activate O-GlcNAcylation of neuronal proteins during glucose deprivation. J Biol Chem 283:13009–13020

    Article  CAS  PubMed  Google Scholar 

  • Cheung WD, Sakabe K, Housley MP, Dias WB, Hart GW (2008) O-linked beta-N-acetylglucosaminyltransferase substrate specificity is regulated by myosin phosphatase targeting and other interacting proteins. J Biol Chem 283:33935–33941

    Article  CAS  PubMed  Google Scholar 

  • Choe KP, Strange K (2007) Evolutionarily conserved WNK and Ste20 kinases are essential for acute volume recovery and survival after hypertonic shrinkage in Caenorhabditis elegans. Am J Physiol Cell Physiol 293:C915–C927

    Article  CAS  PubMed  Google Scholar 

  • Chou TY, Hart GW, Dang CV (1995) c-Myc is glycosylated at threonine 58, a known phosphorylation site and a mutational hot spot in lymphomas. J Biol Chem 270:18961–18965

    Article  CAS  PubMed  Google Scholar 

  • Comer FI, Hart GW (1996) Investigating the role of O-GlcNAc on RNA polymerase II. FASEB J 10(6):A1110–A1119

    Google Scholar 

  • Crampton N, Kodiha M, Shrivastava S, Umar R, Stochaj U (2009) Oxidative stress inhibits nuclear protein export by multiple mechanisms that target FG nucleoporins and Crm1. Mol Biol Cell 20:5106–5116

    Article  CAS  PubMed  Google Scholar 

  • El Messaoudi S, Fabbrizio E, Rodriguez C, Chuchana P, Fauquier L, Cheng D, Theillet C, Vandel L, Bedford MT, Sardet C (2006) Coactivator-associated arginine methyltransferase 1 (CARM1) is a positive regulator of the Cyclin E1 gene. Proc Natl Acad Sci USA 103:13351–13356

    Article  CAS  PubMed  Google Scholar 

  • Fu Y, Subramanya A, Rozansky D, Cohen DM (2006) WNK kinases influence TRPV4 channel function and localization. Am J Physiol Renal Physiol 290:F1305–F1314

    Article  CAS  PubMed  Google Scholar 

  • Fulop N, Marchase RB, Chatham JC (2007a) Role of protein O-linked N-acetyl-glucosamine in mediating cell function and survival in the cardiovascular system. Cardiovasc Res 73:288–297

    Article  PubMed  Google Scholar 

  • Fulop N, Zhang Z, Marchase RB, Chatham JC (2007b) Glucosamine cardioprotection in perfused rat heart associated with increased O-Linked N-acetylglucosamine protein modification and altered p38 activation. Am J Physiol Heart Circ Physiol

  • Fulop N, Zhang Z, Marchase RB, Chatham JC (2007c) Glucosamine cardioprotection in perfused rat hearts associated with increased O-linked N-acetylglucosamine protein modification and altered p38 activation. Am J Physiol Heart Circ Physiol 292:H2227–H2236

    Article  CAS  PubMed  Google Scholar 

  • Gambetta MC, Oktaba K, Muller J (2009) Essential role of the glycosyltransferase sxc/Ogt in polycomb repression. Science 325:93–96

    Article  CAS  PubMed  Google Scholar 

  • Griffith LS, Schmitz B (1995) O-linked N-acetylglucosamine is upregulated in Alzheimer brains. Biochem Biophys Res Commun 213:424–431

    Article  CAS  PubMed  Google Scholar 

  • Griffith LS, Schmitz B (1999) O-linked N-acetylglucosamine levels in cerebellar neurons respond reciprocally to pertubations of phosphorylation. Eur J Biochem 262:824–831

    Article  CAS  PubMed  Google Scholar 

  • Guinez C, Mir AM, Dehennaut V, Cacan R, Harduin-Lepers A, Michalski JC, Lefebvre T (2008) Protein ubiquitination is modulated by O-GlcNAc glycosylation. FASEB J 22:2901–2911

    Article  CAS  PubMed  Google Scholar 

  • Harsha HC, Molina H, Pandey A (2008) Quantitative proteomics using stable isotope labeling with amino acids in cell culture. Nat Protoc 3:505–516

    Article  CAS  PubMed  Google Scholar 

  • Hart GW, Housley MP, Slawson C (2007) Cycling of O-linked beta-N-acetylglucosamine on nucleocytoplasmic proteins. Nature 446:1017–1022

    Article  CAS  PubMed  Google Scholar 

  • Heilig C, Brosius F, Siu B, Concepcion L, Mortensen R, Heilig K, Zhu M, Weldon R, Wu G, Conner D (2003) Implications of glucose transporter protein type 1 (GLUT1)-haplodeficiency in embryonic stem cells for their survival in response to hypoxic stress. Am J Pathol 163:1873–1885

    Article  CAS  PubMed  Google Scholar 

  • Hoorens A, Van de Casteele M, Klöppel G, Pipeleers D (1996) Glucose promotes survival of rat pancreatic beta cells by activating synthesis of proteins which suppress a constitutive apoptotic program. JClin Invest 98:1568–1574

    Article  CAS  Google Scholar 

  • Ibarrola N, Kalume DE, Gronborg M, Iwahori A, Pandey A (2003) A proteomic approach for quantitation of phosphorylation using stable isotope labeling in cell culture. Anal Chem 75:6043–6049

    Article  CAS  PubMed  Google Scholar 

  • Jha S, Dutta A (2009) RVB1/RVB2: running rings around molecular biology. Mol Cell 34:521–533

    Article  CAS  PubMed  Google Scholar 

  • Jones SP, Zachara NE, Ngoh GA, Hill BG, Teshima Y, Bhatnagar A, Hart GW, Marban E (2008) Cardioprotection by N-acetylglucosamine linkage to cellular proteins. Circulation 117:1172–1182

    Article  CAS  PubMed  Google Scholar 

  • Kamemura K, Hart GW (2003) Dynamic interplay between O-glycosylation and O-phosphorylation of nucleocytoplasmic proteins: a new paradigm for metabolic control of signal transduction and transcription. Prog Nucleic Acid Res Mol Biol 73:107–136

    Article  CAS  PubMed  Google Scholar 

  • Kodiha M, Tran D, Morogan A, Qian C, Stochaj U (2009) Dissecting the signaling events that impact classical nuclear import and target nuclear transport factors. PLoS One 4:e8420

    Article  PubMed  Google Scholar 

  • Kulshreshtha R, Ferracin M, Negrini M, Calin GA, Davuluri RV, Ivan M (2007a) Regulation of microRNA expression: the hypoxic component. Cell Cycle 6:1426–1431

    Article  CAS  PubMed  Google Scholar 

  • Kulshreshtha R, Ferracin M, Wojcik SE, Garzon R, Alder H, Agosto-Perez FJ, Davuluri R, Liu CG, Croce CM, Negrini M et al (2007b) A microRNA signature of hypoxia. Mol Cell Biol 27:1859–1867

    Article  CAS  PubMed  Google Scholar 

  • Laczy B, Hill BG, Wang K, Paterson AJ, White CR, Xing D, Chen YF, Darley-Usmar V, Oparil S, Chatham JC (2009) Protein O-GlcNAcylation: a new signaling paradigm for the cardiovascular system. Am J Physiol Heart Circ Physiol 296:H13–H28

    Article  CAS  PubMed  Google Scholar 

  • Langland JO, Kao PN, Jacobs BL (1999) Nuclear factor-90 of activated T-cells: a double-stranded RNA-binding protein and substrate for the double-stranded RNA-dependent protein kinase, PKR. Biochemistry 38:6361–6368

    Article  CAS  PubMed  Google Scholar 

  • Lefebvre T, Alonso C, Mahboub S, Dupire MJ, Zanetta JP, Caillet-Boudin ML, Michalski JC (1999) Effect of okadaic acid on O-linked N-acetylglucosamine levels in a neuroblastoma cell line. Biochim Biophys Acta 1472:71–81

    CAS  PubMed  Google Scholar 

  • Lefebvre T, Ferreira S, Dupont-Wallois L, Bussiere T, Dupire MJ, Delacourte A, Michalski JC, Caillet-Boudin ML (2003) Evidence of a balance between phosphorylation and O-GlcNAc glycosylation of Tau proteins—a role in nuclear localization. Biochim Biophys Acta 1619:167–176

    CAS  PubMed  Google Scholar 

  • Leung AK, Calabrese JM, Sharp PA (2006) Quantitative analysis of Argonaute protein reveals microRNA-dependent localization to stress granules. Proc Natl Acad Sci USA 103:18125–18130

    Article  CAS  PubMed  Google Scholar 

  • Lim KH, Chang HI (2006) O-linked N-acetylglucosamine suppresses thermal aggregation of Sp1. FEBS Lett 580:4645–4652

    Article  CAS  PubMed  Google Scholar 

  • Liu J, Pang Y, Chang T, Bounelis P, Chatham JC, Marchase RB (2006) Increased hexosamine biosynthesis and protein O-GlcNAc levels associated with myocardial protection against calcium paradox and ischemia. J Mol Cell Cardiol 40:303–312

    Article  CAS  PubMed  Google Scholar 

  • Liu J, Marchase RB, Chatham JC (2007) Increased O-GlcNAc levels during reperfusion lead to improved functional recovery and reduced calpain proteolysis. Am J Physiol Heart Circ Physiol 293:H1391–H1399

    Article  CAS  PubMed  Google Scholar 

  • Loberg RD, Vesely E, Brosius FC III (2002) Enhanced glycogen synthase kinase-3beta activity mediates hypoxia-induced apoptosis of vascular smooth muscle cells and is prevented by glucose transport and metabolism. J Biol Chem 277:41667–41673

    Article  CAS  PubMed  Google Scholar 

  • Malhotra R, Brosius FC 3rd (1999) Glucose uptake and glycolysis reduce hypoxia-induced apoptosis in cultured neonatal rat cardiac myocytes. J Biol Chem 274:12567–12575

    Article  CAS  PubMed  Google Scholar 

  • Marsit CJ, Eddy K, Kelsey KT (2006) MicroRNA responses to cellular stress. Cancer Res 66:10843–10848

    Article  CAS  PubMed  Google Scholar 

  • Moley KH, Mueckler MM (2000) Glucose transport and apoptosis. Apoptosis 5:99–105

    Article  CAS  PubMed  Google Scholar 

  • Musicki B, Kramer MF, Becker RE, Burnett AL (2005) Inactivation of phosphorylated endothelial nitric oxide synthase (Ser-1177) by O-GlcNAc in diabetes-associated erectile dysfunction. Proc Natl Acad Sci USA 102:11870–11875

    Article  CAS  PubMed  Google Scholar 

  • Nagy T, Champattanachai V, Marchase RB, Chatham JC (2006) Glucosamine inhibits angiotensin II-induced cytoplasmic Ca2 + elevation in neonatal cardiomyocytes via protein-associated O-linked N-acetylglucosamine. Am J Physiol Cell Physiol 290:C57–C65

    Article  CAS  PubMed  Google Scholar 

  • Ngoh GA, Jones SP (2008) New insights into metabolic signaling and cell survival: the role of beta-O-linkage of N-acetylglucosamine. J Pharmacol Exp Ther 327:602–609

    Article  CAS  PubMed  Google Scholar 

  • Ngoh GA, Watson LJ, Facundo HT, Dillmann W, Jones SP (2008) Non-canonical glycosyltransferase modulates post-hypoxic cardiac myocyte death and mitochondrial permeability transition. J Mol Cell Cardiol 45:313–325

    Article  CAS  PubMed  Google Scholar 

  • Ngoh GA, Facundo HT, Hamid T, Dillmann W, Zachara NE, Jones SP (2009) Unique hexosaminidase reduces metabolic survival signal and sensitizes cardiac myocytes to hypoxia/reoxygenation injury. Circ Res 104:41–49

    Article  CAS  PubMed  Google Scholar 

  • Not LG, Marchase RB, Fulop N, Brocks CA, Chatham JC (2007) Glucosamine administration improves survival rate after severe hemorrhagic shock combined with trauma in rats. Shock

  • Ohn T, Kedersha N, Hickman T, Tisdale S, Anderson P (2008) A functional RNAi screen links O-GlcNAc modification of ribosomal proteins to stress granule and processing body assembly. Nat Cell Biol 10:1224–1231

    Article  CAS  PubMed  Google Scholar 

  • Ong SE, Blagoev B, Kratchmarova I, Kristensen DB, Steen H, Pandey A, Mann M (2002) Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics. Mol Cell Proteomics 1:376–386

    Article  CAS  PubMed  Google Scholar 

  • Pang Y, Hunton DL, Bounelis P, Marchase RB (2002) Hyperglycemia inhibits capacitative calcium entry and hypertrophy in neonatal cardiomyocytes. Diabetes 51:3461–3467

    Article  CAS  PubMed  Google Scholar 

  • Phillips JR, Dalmay T, Bartels D (2007) The role of small RNAs in abiotic stress. FEBS Lett 581:3592–3597

    Article  CAS  PubMed  Google Scholar 

  • Ren XP, Wu J, Wang X, Sartor MA, Qian J, Jones K, Nicolaou P, Pritchard TJ, Fan GC (2009) MicroRNA-320 is involved in the regulation of cardiac ischemia/reperfusion injury by targeting heat-shock protein 20. Circulation 119:2357–2366

    Article  CAS  PubMed  Google Scholar 

  • Reyes JL, Arenas-Huertero C, Sunkar R (2010) Cloning of stress-responsive microRNAs and other small RNAs from plants. Methods Mol Biol 639:239–251

    Article  CAS  PubMed  Google Scholar 

  • Sakamoto S, Aoki K, Higuchi T, Todaka H, Morisawa K, Tamaki N, Hatano E, Fukushima A, Taniguchi T, Agata Y (2009) The NF90-NF45 complex functions as a negative regulator in the microRNA processing pathway. Mol Cell Biol 29:3754–3769

    Article  CAS  PubMed  Google Scholar 

  • Schwartz YB, Pirrotta V (2009) A little bit of sugar makes polycomb better. J Mol Cell Biol 1:11–12

    Article  CAS  PubMed  Google Scholar 

  • Scoumanne A, Chen X (2008) Protein methylation: a new mechanism of p53 tumor suppressor regulation. Histol Histopathol 23:1143–1149

    CAS  PubMed  Google Scholar 

  • Shi L, Qiu D, Zhao G, Corthesy B, Lees-Miller S, Reeves WH, Kao PN (2007) Dynamic binding of Ku80, Ku70 and NF90 to the IL-2 promoter in vivo in activated T-cells. Nucleic Acids Res 35:2302–2310

    Article  CAS  PubMed  Google Scholar 

  • Sinclair DA, Syrzycka M, Macauley MS, Rastgardani T, Komljenovic I, Vocadlo DJ, Brock HW, Honda BM (2009) Drosophila O-GlcNAc transferase (OGT) is encoded by the Polycomb group (PcG) gene, super sex combs (sxc). Proc Natl Acad Sci USA 106:13427–13432

    Article  CAS  PubMed  Google Scholar 

  • Singleton KD, Wischmeyer PE (2008) Glutamine induces heat shock protein expression via O-glycosylation and phosphorylation of HSF-1 and Sp1. JPEN J Parenter Enteral Nutr 32:371–376

    Article  CAS  PubMed  Google Scholar 

  • Sohn KC, Lee KY, Park JE, Do SI (2004) OGT functions as a catalytic chaperone under heat stress response: a unique defense role of OGT in hyperthermia. Biochem Biophys Res Commun 322:1045–1051

    Article  CAS  PubMed  Google Scholar 

  • Sunkar R (2010) MicroRNAs with macro effects on plant stress responses. Semin Cell Dev Biol

  • Tallent MK, Varghis N, Skorobogatko Y, Hernandez-Cuebas L, Whelan K, Vocadlo DJ, Vosseller K (2009) In vivo modulation of O-GlcNAc levels regulates hippocampal synaptic plasticity through interplay with phosphorylation. J Biol Chem 284:174–181

    Article  CAS  PubMed  Google Scholar 

  • Tang J, Kao PN, Herschman HR (2000) Protein-arginine methyltransferase I, the predominant protein-arginine methyltransferase in cells, interacts with and is regulated by interleukin enhancer-binding factor 3. J Biol Chem 275:19866–19876

    Article  CAS  PubMed  Google Scholar 

  • Tang Y, Zheng J, Sun Y, Wu Z, Liu Z, Huang G (2009) MicroRNA-1 regulates cardiomyocyte apoptosis by targeting Bcl-2. Int Heart J 50:377–387

    Article  CAS  PubMed  Google Scholar 

  • Teo CF, Ingale S, Wolfert MA, Elsayed GA, Not LG, Chatham JC, Wells L, Boons GJ (2010) Glycopeptide-specific monoclonal antibodies suggest new roles for O-GlcNAc. Nat Chem Biol 6:338–343

    Article  CAS  PubMed  Google Scholar 

  • Ting NS, Kao PN, Chan DW, Lintott LG, Lees-Miller SP (1998) DNA-dependent protein kinase interacts with antigen receptor response element binding proteins NF90 and NF45. J Biol Chem 273:2136–2145

    Article  CAS  PubMed  Google Scholar 

  • van Rooij E, Sutherland LB, Qi X, Richardson JA, Hill J, Olson EN (2007) Control of stress-dependent cardiac growth and gene expression by a microRNA. Science 316:575–579

    Article  PubMed  Google Scholar 

  • Verissimo F, Silva E, Morris JD, Pepperkok R, Jordan P (2006) Protein kinase WNK3 increases cell survival in a caspase-3-dependent pathway. Oncogene 25:4172–4182

    Article  CAS  PubMed  Google Scholar 

  • Vitari AC, Deak M, Morrice NA, Alessi DR (2005) The WNK1 and WNK4 protein kinases that are mutated in Gordon’s hypertension syndrome phosphorylate and activate SPAK and OSR1 protein kinases. Biochem J 391:17–24

    Article  CAS  PubMed  Google Scholar 

  • Wang Z, Pandey A, Hart GW (2007) Dynamic interplay between O-linked N-acetylglucosaminylation and glycogen synthase kinase-3-dependent phosphorylation. Mol Cell Proteomics 6:1365–1379

    Article  CAS  PubMed  Google Scholar 

  • Wang Z, Gucek M, Hart GW (2008) Cross-talk between GlcNAcylation and phosphorylation: site-specific phosphorylation dynamics in response to globally elevated O-GlcNAc. Proc Natl Acad Sci USA 105:13793–13798

    Article  CAS  PubMed  Google Scholar 

  • Wang Z, Udeshi ND, O’Malley M, Shabanowitz J, Hunt DF, Hart GW (2010a) Enrichment and site mapping of O-linked N-acetylglucosamine by a combination of chemical/enzymatic tagging, photochemical cleavage, and electron transfer dissociation mass spectrometry. Mol Cell Proteomics 9:153–160

    Article  CAS  PubMed  Google Scholar 

  • Wang Z, Udeshi ND, Slawson C, Compton PD, Sakabe K, Cheung WD, Shabanowitz J, Hunt DF, Hart GW (2010b) Extensive crosstalk between O-GlcNAcylation and phosphorylation regulates cytokinesis. Sci Signal 3, ra2

  • Wilson FH, Disse-Nicodeme S, Choate KA, Ishikawa K, Nelson-Williams C, Desitter I, Gunel M, Milford DV, Lipkin GW, Achard JM et al (2001) Human hypertension caused by mutations in WNK kinases. Science 293:1107–1112

    Article  CAS  PubMed  Google Scholar 

  • Wysocka J, Allis CD, Coonrod S (2006) Histone arginine methylation and its dynamic regulation. Front Biosci 11:344–355

    Article  CAS  PubMed  Google Scholar 

  • Xu P, Vernooy SY, Guo M, Hay BA (2003) The Drosophila microRNA Mir-14 suppresses cell death and is required for normal fat metabolism. Curr Biol 13:790–795

    Article  CAS  PubMed  Google Scholar 

  • Xu BE, Stippec S, Lenertz L, Lee BH, Zhang W, Lee YK, Cobb MH (2004) WNK1 activates ERK5 by an MEKK2/3-dependent mechanism. J Biol Chem 279:7826–7831

    Article  CAS  PubMed  Google Scholar 

  • Xu BE, Lee BH, Min X, Lenertz L, Heise CJ, Stippec S, Goldsmith EJ, Cobb MH (2005) WNK1: analysis of protein kinase structure, downstream targets, and potential roles in hypertension. Cell Res 15:6–10

    Article  PubMed  Google Scholar 

  • Xu CF, Yu CH, Li YM (2009) Regulation of hepatic microRNA expression in response to ischemic preconditioning following ischemia/reperfusion injury in mice. OMICS 13:513–520

    Article  CAS  PubMed  Google Scholar 

  • Yang S, Zou LY, Bounelis P, Chaudry I, Chatham JC, Marchase RB (2006) Glucosamine administration during resuscitation improves organ function after trauma hemorrhage. Shock 25:600–607

    Article  CAS  PubMed  Google Scholar 

  • Yeo WM, Chow VT (2007) The VP1 structural protein of enterovirus 71 interacts with human ornithine decarboxylase and gene trap ankyrin repeat. Microb Pathog 42:129–137

    Article  CAS  PubMed  Google Scholar 

  • Yin C, Wang X, Kukreja RC (2008) Endogenous microRNAs induced by heat-shock reduce myocardial infarction following ischemia-reperfusion in mice. FEBS Lett 582:4137–4142

    Article  CAS  PubMed  Google Scholar 

  • Yin C, Salloum FN, Kukreja RC (2009) A novel role of microRNA in late preconditioning: upregulation of endothelial nitric oxide synthase and heat shock protein 70. Circ Res 104:572–575

    Article  CAS  PubMed  Google Scholar 

  • Zachara NE, Hart GW (2004a) O-GlcNAc a sensor of cellular state: the role of nucleocytoplasmic glycosylation in modulating cellular function in response to nutrition and stress. Biochim Biophys Acta 1673:13–28

    CAS  PubMed  Google Scholar 

  • Zachara NE, Hart GW (2004b) O-GlcNAc a sensor of cellular state: the role of nucleocytoplasmic glycosylation in modulating cellular functions in response to nutrition and stress. Biochim Biophys Acta Gen Subj (in press)

  • Zachara NE, Hart GW (2006) Cell signaling, the essential role of O-GlcNAc!. Biochim Biophys Acta 1761:599–617

    CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Zagorska A, Pozo-Guisado E, Boudeau J, Vitari AC, Rafiqi FH, Thastrup J, Deak M, Campbell DG, Morrice NA, Prescott AR et al (2007) Regulation of activity and localization of the WNK1 protein kinase by hyperosmotic stress. J Cell Biol 176:89–100

    Article  CAS  PubMed  Google Scholar 

  • Zou L, Yang S, Hu S, Chaudry IH, Marchase RB, Chatham JC (2007) The protective effects of PUGNAc on cardiac function after trauma-hemorrhage are mediated via increased protein O-GlcNAc levels. Shock 27:402–408

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors acknowledge the generous gift of antibodies by Dr. Peter N. Kao (Anti-NF-90 and NF-45), Dr. Jean-Pierre Paccaud (Anti-Sec24), Dr. Katsuko Tani (Anti-p125i), and Dr. Taka-aki Tamura (anti-Tip49α, antiTip49β). The author’s work is supported by NIH grants HD R37-13563, CA R01-42486, DK-R01-61671, DK-R21/33-71280 (GWH), the National Heart, Lung, and Blood Institute, National Institutes of Health, contract No. N01-HV-28180 (GWH and AP), an NIH roadmap grant for Technology Centers of Networks and Pathways (U54RR020839), and an A*Star Research Grant to Johns Hopkins Singapore (NEZ). Under a licensing agreement between Covance Research Products and The Johns Hopkins University, Dr. Hart receives a share of royalties received by the university on sales of the CTD 110.6 antibody. The terms of this arrangement are being managed by The Johns Hopkins University in accordance with its conflict of interest policies.

Author information

Authors and Affiliations

  1. The Department of Biological Chemistry, The Johns Hopkins University School of Medicine, 725 N. Wolfe Street, Baltimore, MD, 21205-2185, USA

    Natasha E. Zachara, Henrik Molina, Akhilesh Pandey & Gerald W. Hart

  2. The Division of Biomedical Sciences, Johns Hopkins Singapore, 31 Biopolis Way, The Nanos #02-01, Singapore, 138669, Singapore

    Natasha E. Zachara & Ker Yi Wong

  3. McKusick-Nathans Institute for Genetic Medicine, Department of Biological Chemistry and Oncology, Johns Hopkins University, 733 N. Broadway, Baltimore, MD, 21205, USA

    Henrik Molina & Akhilesh Pandey

Authors
  1. Natasha E. Zachara
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Henrik Molina
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ker Yi Wong
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Akhilesh Pandey
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Gerald W. Hart
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Natasha E. Zachara.

Electronic supplementary material

Below is the link to the electronic supplementary material.

(PDF 44 kb)

Supplementary Table 2 Raw Mass Spectrometry Data (XLS 179 kb)

726_2010_695_MOESM3_ESM.eps

Supplementary Fig. S1 NF45 does not appear to be O-GlcNAc modified. NF-90 was immunoprecipitated from Cos-7 cells and separated by SDS–PAGE (16.5cm, 7.5% gel). While NF-90 appeared in the complex, we could not detect it by blotting with CTD110.6 suggesting that it’s occurance in the CTD110.6 IP is due to the other O-GlcNAcylated proteins in this complex (EPS 4467 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zachara, N.E., Molina, H., Wong, K.Y. et al. The dynamic stress-induced “O-GlcNAc-ome” highlights functions for O-GlcNAc in regulating DNA damage/repair and other cellular pathways. Amino Acids 40, 793–808 (2011). https://doi.org/10.1007/s00726-010-0695-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00726-010-0695-z

Keywords

Search

Navigation