O-Linked GlcNAc Transferase Is a Conserved Nucleocytoplasmic Protein Containing Tetratricopeptide Repeats

Lubas, William A.; Frank, David W.; Krause, Michael; Hanover, John A.

doi:10.1074/jbc.272.14.9316

Cited by 479 publications

(452 citation statements)

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“…Unlike many other glycosyltransferases, OGT is a soluble protein that is predominantly localized to the nucleus, mitochondria, and cytoplasm of all tissues studied thus far (Kreppel et al 1997;Haltiwanger et al 1992;Lubas et al 1997;Love et al 2003;Hanover et al 2003). Two functional domains characterize OGT: an N-terminal tetratricopeptide repeat (TPR domain) and a C-terminal catalytic domain belonging to the glycogen phosphorylase superfamily (Kreppel et al 1997;Lubas et al 1997;Wrabl and Grishin 2001).…”

Section: O-glcnac Transferasementioning

confidence: 99%

“…Two functional domains characterize OGT: an N-terminal tetratricopeptide repeat (TPR domain) and a C-terminal catalytic domain belonging to the glycogen phosphorylase superfamily (Kreppel et al 1997;Lubas et al 1997;Wrabl and Grishin 2001). Recently, the structure of OGT and the TPR domain have been solved (Lazarus et al 2011;Martinez-Fleites et al 2008;Jinek et al 2004).…”

Section: O-glcnac Transferasementioning

confidence: 99%

“…The gene for OGT maps to Xq13 on the mammalian X chromosome and encodes three well-characterized variants: short OGT, mitochondrial OGT (mOGT), and nuclear/cytoplasmic OGT (Kreppel et al 1997;Lubas et al 1997;Love et al 2003;Hanover et al 2003). mOGT has an alternative mitochondrial targeting sequence on its N-terminus.…”

Section: O-glcnac Transferasementioning

confidence: 99%

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Dynamic O-GlcNAcylation and its roles in the cellular stress response and homeostasis

Groves

Lee

Yildirir

et al. 2013

Cell Stress and Chaperones

119

112

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O-linked N-acetyl-β-D-glucosamine (O-GlcNAc) is a ubiquitous and dynamic post-translational modification known to modify over 3,000 nuclear, cytoplasmic, and mitochondrial eukaryotic proteins. Addition of O-GlcNAc to proteins is catalyzed by the O-GlcNAc transferase and is removed by a neutral-N-acetyl-β-glucosaminidase (OGlcNAcase). O-GlcNAc is thought to regulate proteins in a manner analogous to protein phosphorylation, and the cycling of this carbohydrate modification regulates many cellular functions such as the cellular stress response. Diverse forms of cellular stress and tissue injury result in enhanced O-GlcNAc modification, or O-GlcNAcylation, of numerous intracellular proteins. Stress-induced OGlcNAcylation appears to promote cell/tissue survival by regulating a multitude of biological processes including: the phosphoinositide 3-kinase/Akt pathway, heat shock protein expression, calcium homeostasis, levels of reactive oxygen species, ER stress, protein stability, mitochondrial dynamics, and inflammation. Here, we will discuss the regulation of these processes by O-GlcNAc and the impact of such regulation on survival in models of ischemia reperfusion injury and trauma hemorrhage. We will also discuss the misregulation of O-GlcNAc in diseases commonly associated with the stress response, namely Alzheimer's and Parkinson's diseases. Finally, we will highlight recent advancements in the tools and technologies used to study the O-GlcNAc modification.

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Section: O-glcnac Transferasementioning

confidence: 99%

Section: O-glcnac Transferasementioning

confidence: 99%

See 1 more Smart Citation

Dynamic O-GlcNAcylation and its roles in the cellular stress response and homeostasis

Groves

Lee

Yildirir

et al. 2013

Cell Stress and Chaperones

119

112

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“…Rat CREB and human OGT cDNA clones were generously provided by Dr. R. H. Goodman (Oregon Health & Science University) 4 and Dr. J. A. Hanover (NIDDK, National Institutes of Health), 5 respectively, and were cloned into baculovirus expression vectors in frame with a histidine tag. Baculovirus preparation and protein expression in Sf9 cells were performed by Dr. P. Snow at the Beckman Institute Protein Expression Facility at the California Institute of Technology.…”

mentioning

confidence: 99%

Dynamic Glycosylation of the Transcription Factor CREB: A Potential Role in Gene Regulation

2003

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Experimental Section. General. Unless otherwise noted, reagents were purchased from the commercial suppliers Fisher (Fairlawn, NJ) and Sigma-Aldrich (St. Louis, MO) and were used without further purification. Protease and phosphatase inhibitors were purchased from Sigma-Aldrich or Alexis Biochemicals (San Diego, CA). Bovine GalT and ovalbumin were obtained from Sigma-Aldrich. Preparation of rat brain nuclear extracts. Nuclear extracts were prepared as previously reported 1 with minor modifications. The forebrains of Sprague Dawley rats (Charles River Laboratories, Kingston, MA) were dissected on ice and homogenized in 10 volumes of ice-cold buffer A (10 mM HEPES pH 7.9, 1.5 mM MgCl 2 , 10 mM KCl, 0.5 mM DTT) containing protease inhibitors (5 µg/ml pepstatin, 5 µg/ml chymostatin, 20 µg/ml leupeptin, 20 µg/ml aprotinin, 20 µg/ml antipain, 0.2 mM PMSF), phosphatase inhibitors (20 mM NaF, 1 mM Na 3 VO 4 , 50 µM Na 2 MoO 4 ), and hexosaminidase inhibitors (50 mM GlcNAc and 10 µM streptozocin). The resulting lysate was centrifuged at 1,000xg at 4 o C for 10 min, and the crude nuclear pellet was washed with buffer A. The pellet was resuspended at 2 mg/ml in buffer B (20 mM HEPES pH 7.9, 0.42 M NaCl, 1.5 mM MgCl 2 , 0.2 mM EDTA, 50 mM GlcNAc, 0.2 mM PMSF, 25% (v/v) glycerol) by stirring at 4 o C for 40 min. Following centrifugation at 10,000xg for 30 min, the supernatant was defined as the nuclear extract. The nuclear extract was dialyzed against buffer D (20 mM HEPES pH 7.9, 0.1 M KCl, 0.5 mM DTT, 0.2 mM EDTA, 50 mM GlcNAc, 20% (v/v) glycerol) for 6 h at 4 o C, and proteins were precipitated using (NH 4 ) 2 SO 4 (38% final concentration). The protein pellet obtained after centrifugation at 21,500xg for 10 min was solubilized in buffer C (0.3 volumes; 25 mM Tris pH7.5, 1 mM EDTA, 1 mM DTT containing protease inhibitors) by gentle mixing for 1 h at 4 o C. Following clarification at 21,500xg for 5 min, the supernatant was dialyzed into buffer G (20 mM HEPES pH 7.3, 0.1 M KCl, 0.5 mM DTT, 0.2 mM EDTA, 5 mM MnCl 2 , 0.2 mM PMSF) overnight at 4 o C. The protein concentration was determined using the BCA assay (Pierce/Endogen Biotech, Rockford, IL).

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“…De fait, on sait maintenant que la O-GlcNAcylation est une modification réversible qui, de façon analogue à la phosphorylation, contrôle l'activité des protéines et les interactions entre protéines au sein d'assemblages moléculaires [6]. Cependant, contrairement aux phosphorylations/déphosphorylations qui sont régulées par une myriade de kinases et phosphatases, deux enzymes seulement, l'OGT (O-linked N-acetyl-glucosaminyltransferase) et la N-Acétyl--D glucosaminidase (O-GlcNAcase ou OGA), dont les gènes ont été clonés respectivement en 1997 [7,8] et 2001 [9], contrôlent le niveau de O-GlcNAcylation des protéines (Figure 1). Bien que cette modification soit connue depuis maintenant un quart de siècle, la communauté scientifique commence tout juste à prendre conscience de son importance dans la plupart des processus biologiques.…”

Section: La O-glcnacylation Modifie L'activité Des Protéinesunclassified

O-GlcNAc glycosylation et régulation de la signalisation cellulaire

Issad

2010

Med Sci (Paris)

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Le séquençage du génome humain, au début des années 2000, a révélé que notre patrimoine génétique comprenait moins de 40 000 gènes (20-25 000 gènes codant pour des protéines selon des estimations plus récentes [1]) alors que les cellules vivantes sont des systèmes sophistiqués comprenant un nombre très important d'activités biologiques complexes. Cette grande diversité des activités cellulaires, surprenante au regard du faible nombre de gènes qui codent pour les protéines, peut en partie s'expliquer par les nombreuses modifications post-traductionnelles (phosphorylation, glycosylation, sumoylation, acétylation, ubiquitinylation, nitrosylation, palmitoylation, farnésylation, méthyla-tion, ADP-ribosylation, hydroxylation, oxydation, etc.) qui augmentent de façon importante les potentialités fonctionnelles des protéines. Ces modifications contrô-lent en particulier leur localisation dans différents compartiments cellulaires et leur interaction avec différents partenaires au sein d'assemblages multimoléculaires, permettant la mise en place de réseaux de signalisation complexes. Les phosphorylations et glycosylations constituent les modifications post-traductionnelles les plus abondantes et les mieux connues. Cependant, alors que les phosphorylations sont généralement considérées comme étant des modifications rapides et réversibles, permettant à la cellule de répondre à divers stimulus et de s'adapter aux variations de son environnement, les glycosylations ont longtemps été considérées comme des modifications stables, impliquant l'ajout de chaî-nes complexes d'hydrates de carbone, qui généralement restent présentes sur la protéine mature tout au long de son existence. Ces glycosylations complexes, sur des résidus sérine/thréonine (O-glycosylations) ou asparagine (N-glycosylations), ne se rencontrent que dans certains compartiments cellulaires (réticulum endoplasmique, appareil de Golgi, face externe de la membrane plasmique) et concernent essentiellement les protéines membranaires ou sécrétées. Cependant, en 1984, en étudiant la distribution des résidus N-Acétyl-glucosamine terminaux à la surface des lymphocytes T, G. W. Hart [2] a découvert un type de glycosylation original, la O-N-Acétylglucosaminylation (O-GlcNAcylation), qui consiste en l'addition d'un monosaccharide, la N-acétyl-glucosamine, sur le groupement hydroxyl d'acides aminés sérine ou thréonine (Figure 1). Il s'est ensuite rendu compte que, contrairement aux glycosylations « classiques », cette modification était observée essentiellement dans le cytoplasme et le noyau de la cellule [3,4]. > La O-GlcNAcylation correspond à l'addition de N-Acétylglucosamine sur des résidus sérine/thréo-nine de protéines cytosoliques ou nucléaires. Elle constitue un mode de régulation post-traductionnel réversible, analogue aux phosphorylations, qui contrôle l'activité, la stabilité ou la localisation des protéines en fonction de la disponibilité en glucose. Des perturbations de la O-GlcNAcylation des protéines pourraient jouer un rôle important dans des pathologies humain...

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O-Linked GlcNAc Transferase Is a Conserved Nucleocytoplasmic Protein Containing Tetratricopeptide Repeats

Cited by 479 publications

References 35 publications

Dynamic O-GlcNAcylation and its roles in the cellular stress response and homeostasis

Dynamic O-GlcNAcylation and its roles in the cellular stress response and homeostasis

Dynamic Glycosylation of the Transcription Factor CREB: A Potential Role in Gene Regulation

O-GlcNAc glycosylation et régulation de la signalisation cellulaire

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