Abstract
O-GlcNAc-ylation is the post-translational addition of an O-linked β-N-acetylglucosamine to the serine and threonine residues of thousands of proteins in eukaryotic cells. Specifically, half of the thirty different types of protein components in the nuclear pore complex (NPC) are modified by O-GlcNAc, of which the majority are intrinsically disordered nucleoporins (Nups) containing multiple phenylalanine-glycine (FG) repeats. Moreover, these FG-Nups form a strict selectivity barrier with a high density of O-GlcNAc in the NPC to mediate bidirectional trafficking between the cytoplasm and nucleus. However, the roles that O-GlcNAc plays in the structure and function of the NPC remain obscure. In this review paper, we will discuss the current knowledge of O-GlcNAc-ylated Nups, highlight some new techniques used to probe O-GlcNAc’s roles in the nuclear pore, and finally propose a new model for the effect of O-GlcNAc on the NPC’s permeability.
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References
Akey, C. W., and M. Radermacher. Architecture of the Xenopus nuclear pore complex revealed by three-dimensional cryo-electron microscopy. J. Cell Biol. 122(1):1–19, 1993.
Beck, M., F. Forster, et al. Nuclear pore complex structure and dynamics revealed by cryoelectron tomography. Science 306(5700):1387–1390, 2004.
Blatch, G. L., and M. Lassle. The tetratricopeptide repeat: a structural motif mediating protein-protein interactions. BioEssays 21(11):932–939, 1999.
Bond, M. R., and J. A. Hanover. A little sugar goes a long way: the cell biology of O-GlcNAc. J. Cell Biol. 208(7):869–880, 2015.
Buse, M. G. Hexosamines, insulin resistance, and the complications of diabetes: current status. Am. J. Physiol. Endocrinol. Metab. 290(1):E1–E8, 2006.
Butkinaree, C., K. Park, et al. O-linked beta-N-acetylglucosamine (O-GlcNAc): extensive crosstalk with phosphorylation to regulate signaling and transcription in response to nutrients and stress. Biochim. Biophys. Acta 1800(2):96–106, 2010.
Chikanishi, T., R. Fujiki, et al. Glucose-induced expression of MIP-1 genes requires O-GlcNAc transferase in monocytes. Biochem. Biophys. Res. Commun. 394(4):865–870, 2010.
Chou, T. Y., and G. W. Hart. O-linked N-acetylglucosamine and cancer: messages from the glycosylation of c-Myc. Adv. Exp. Med. Biol. 491:413–418, 2001.
Chou, C. F., A. J. Smith, et al. Characterization and dynamics of O-linked glycosylation of human cytokeratin 8 and 18. J. Biol. Chem. 267(6):3901–3906, 1992.
Clark, P. M., J. F. Dweck, et al. Direct in-gel fluorescence detection and cellular imaging of O-GlcNAc-modified proteins. J. Am. Chem. Soc. 130(35):11576–11577, 2008.
Copeland, R. J., J. W. Bullen, et al. Cross-talk between GlcNAcylation and phosphorylation: roles in insulin resistance and glucose toxicity. Am. J. Physiol. Endocrinol. Metab. 295(1):E17–E28, 2008.
Crampton, N., M. Kodiha, et al. Oxidative stress inhibits nuclear protein export by multiple mechanisms that target FG nucleoporins and Crm1. Mol. Biol. Cell 20(24):5106–5116, 2009.
D’Angelo, M. A., and M. W. Hetzer. Structure, dynamics and function of nuclear pore complexes. Trends Cell Biol. 18(10):456–466, 2008.
Dieterich, D. C., and M. R. Kreutz. Proteomics of the synapse—a quantitative approach to neuronal plasticity. Mol. Cell. Proteomics, p. mcp-R115, 2015.
Favreau, C., H. J. Worman, et al. Cell cycle-dependent phosphorylation of nucleoporins and nuclear pore membrane protein Gp210. Biochemistry 35(24):8035–8044, 1996.
Finlay, D. R., and D. J. Forbes. Reconstitution of biochemically altered nuclear pores: transport can be eliminated and restored. Cell 60(1):17–29, 1990.
Frey, S., and D. Gorlich. A saturated FG-repeat hydrogel can reproduce the permeability properties of nuclear pore complexes. Cell 130(3):512–523, 2007.
Gambetta, M. C., K. Oktaba, et al. Essential role of the glycosyltransferase sxc/Ogt in polycomb repression. Science 325(5936):93–96, 2009.
Goldberg, M. W., and T. D. Allen. High resolution scanning electron microscopy of the nuclear envelope: demonstration of a new, regular, fibrous lattice attached to the baskets of the nucleoplasmic face of the nuclear pores. J. Cell Biol. 119(6):1429–1440, 1992.
Gupta, R., and S. Brunak. Prediction of glycosylation across the human proteome and the correlation to protein function. Pac. Symp. Biocomput. 20022002:310–322, 2002.
Gut, P., and E. Verdin. The nexus of chromatin regulation and intermediary metabolism. Nature 502(7472):489–498, 2013.
Hanover, J. A., C. K. Cohen, et al. O-linked N-acetylglucosamine is attached to proteins of the nuclear pore. Evidence for cytoplasmic and nucleoplasmic glycoproteins. J. Biol. Chem. 262(20):9887–9894, 1987.
Hanover, J. A., S. Yu, et al. Mitochondrial and nucleocytoplasmic isoforms of O-linked GlcNAc transferase encoded by a single mammalian gene. Arch. Biochem. Biophys. 409(2):287–297, 2003.
Hart, G. W., M. P. Housley, et al. Cycling of O-linked beta-N-acetylglucosamine on nucleocytoplasmic proteins. Nature 446(7139):1017–1022, 2007.
Harwood, K. R., and J. A. Hanover. Nutrient-driven O-GlcNAc cycling—think globally but act locally. J. Cell Sci. 127(9):1857–1867, 2014.
Hicks, G. R., and N. V. Raikhel. Protein import into the nucleus: an integrated view. Annu. Rev. Cell Dev. Biol. 11:155–188, 1995.
Hinshaw, J. E., B. O. Carragher, et al. Architecture and design of the nuclear pore complex. Cell 69(7):1133–1141, 1992.
Holt, G. D., C. M. Snow, et al. Nuclear pore complex glycoproteins contain cytoplasmically disposed O-linked N-acetylglucosamine. J. Cell Biol. 104(5):1157–1164, 1987.
Hulsmann, B. B., A. A. Labokha, et al. The permeability of reconstituted nuclear pores provides direct evidence for the selective phase model. Cell 150(4):738–751, 2012.
Jia, C. Z., T. Liu, et al. O-GlcNAcPRED: a sensitive predictor to capture protein O-GlcNAcylation sites. Mol. Biosyst. 911:2909–2913, 2013.
Jinek, M., J. Rehwinkel, et al. The superhelical TPR-repeat domain of O-linked GlcNAc transferase exhibits structural similarities to importin alpha. Nat. Struct. Mol. Biol. 11(10):1001–1007, 2004.
Kim, E. J., L. K. Abramowitz, et al. Versatile O-GlcNAc transferase assay for high-throughput identification of enzyme variants, substrates, and inhibitors. Bioconjug. Chem. 25(6):1025–1030, 2014.
Kodiha, M., A. Chu, et al. Multiple mechanisms promote the inhibition of classical nuclear import upon exposure to severe oxidative stress. Cell Death Differ. 11(8):862–874, 2004.
Kodiha, M., N. Crampton, et al. Traffic control at the nuclear pore. Nucleus 1(3):237–244, 2010.
Kodiha, M., D. Tran, et al. Dissecting the signaling events that impact classical nuclear import and target nuclear transport factors. PLoS One 4(12):e8420, 2009.
Kreppel, L. K., M. A. Blomberg, et al. Dynamic glycosylation of nuclear and cytosolic proteins. Cloning and characterization of a unique O-GlcNAc transferase with multiple tetratricopeptide repeats. J. Biol. Chem. 272(14):9308–9315, 1997.
Labokha, A. A., S. Gradmann, et al. Systematic analysis of barrier-forming FG hydrogels from Xenopus nuclear pore complexes. EMBO J. 32(2):204–218, 2013.
Laczy, B., B. G. Hill, et al. Protein O-GlcNAcylation: a new signaling paradigm for the cardiovascular system. Am. J. Physiol. Heart Circ. Physiol. 296(1):H13–H28, 2009.
Lazarus, B. D., D. C. Love, et al. O-GlcNAc cycling: implications for neurodegenerative disorders. Int. J. Biochem. Cell Biol. 41(11):2134–2146, 2009.
Letschert, S., A. Gohler, et al. Super-resolution imaging of plasma membrane glycans. Angew. Chem. Int. Ed. Engl. 53(41):10921–10924, 2014.
Li, B., and J. J. Kohler. Glycosylation of the nuclear pore. Traffic 15(4):347–361, 2014.
Love, D. C., J. Kochan, et al. Mitochondrial and nucleocytoplasmic targeting of O-linked GlcNAc transferase. J. Cell Sci. 116(4):647–654, 2003.
Ma, J., A. Goryaynov, et al. Self-regulated viscous channel in the nuclear pore complex. Proc. Natl. Acad. Sci. USA 109(19):7326–7331, 2012.
Ma, J., and G. W. Hart. O-GlcNAc profiling: from proteins to proteomes. Clin. Proteomics 11(1):8, 2014.
Ma, J., and W. Yang. Three-dimensional distribution of transient interactions in the nuclear pore complex obtained from single-molecule snapshots. Proc. Natl. Acad. Sci. USA 107(16):7305–7310, 2010.
Macauley, M. S., Y. He, et al. Inhibition of O-GlcNAcase using a potent and cell-permeable inhibitor does not induce insulin resistance in 3T3-L1 adipocytes. Chem. Biol. 17(9):937–948, 2010.
Miller, M. W., and J. A. Hanover. Functional nuclear pores reconstituted with beta 1-4 galactose-modified O-linked N-acetylglucosamine glycoproteins. J. Biol. Chem. 269(12):9289–9297, 1994.
Ozcan, S., S. S. Andrali, et al. Modulation of transcription factor function by O-GlcNAc modification. Biochim. Biophys. Acta 1799(5–6):353–364, 2010.
Park, M. K., M. D’Onofrio, et al. A monoclonal antibody against a family of nuclear pore proteins (nucleoporins): O-linked N-acetylglucosamine is part of the immunodeterminant. Proc. Natl. Acad. Sci. USA 84(18):6462–6466, 1987.
Parker, G., R. Taylor, et al. Hyperglycemia and inhibition of glycogen synthase in streptozotocin-treated mice: role of O-linked N-acetylglucosamine. J. Biol. Chem. 279(20):20636–20642, 2004.
Powers, M. A., C. Macaulay, et al. Reconstituted nuclei depleted of a vertebrate GLFG nuclear pore protein, p97, import but are defective in nuclear growth and replication. J. Cell Biol. 128(5):721–736, 1995.
Radermacher, P. T., F. Myachina, et al. O-GlcNAc reports ambient temperature and confers heat resistance on ectotherm development. Proc. Natl. Acad. Sci. USA 111(15):5592–5597, 2014.
Reeves, R. A., A. Lee, et al. Characterization of the specificity of O-GlcNAc reactive antibodies under conditions of starvation and stress. Anal. Biochem. 457:8–18, 2014.
Ribbeck, K., and D. Gorlich. Kinetic analysis of translocation through nuclear pore complexes. EMBO J. 20(6):1320–1330, 2001.
Ribbeck, K., and D. Gorlich. The permeability barrier of nuclear pore complexes appears to operate via hydrophobic exclusion. EMBO J. 21(11):2664–2671, 2002.
Roquemore, E. P., M. R. Chevrier, et al. Dynamic O-GlcNAcylation of the small heat shock protein alpha B-crystallin. Biochemistry 35(11):3578–3586, 1996.
Sinclair, D. A., M. Syrzycka, et al. Drosophila O-GlcNAc transferase (OGT) is encoded by the Polycomb group (PcG) gene, super sex combs (sxc). Proc. Natl. Acad. Sci. USA 106(32):13427–13432, 2009.
Slawson, C., T. Lakshmanan, et al. A mitotic GlcNAcylation/phosphorylation signaling complex alters the posttranslational state of the cytoskeletal protein vimentin. Mol. Biol. Cell 19(10):4130–4140, 2008.
Snow, C. M., A. Senior, et al. Monoclonal antibodies identify a group of nuclear pore complex glycoproteins. J. Cell Biol. 104(5):1143–1156, 1987.
Sterne-Marr, R., J. M. Blevitt, et al. O-linked glycoproteins of the nuclear pore complex interact with a cytosolic factor required for nuclear protein import. J. Cell Biol. 116(2):271–280, 1992.
Taylor, R. P., G. J. Parker, et al. Glucose deprivation stimulates O-GlcNAc modification of proteins through up-regulation of O-linked N-acetylglucosaminyltransferase. J. Biol. Chem. 283(10):6050–6057, 2008.
Trinidad, J. C., D. T. Barkan, et al. Global identification and characterization of both O-GlcNAcylation and phosphorylation at the murine synapse. Mol. Cell. Proteomics 11(8):215–229, 2012.
van de Linde, S., A. Loschberger, et al. Direct stochastic optical reconstruction microscopy with standard fluorescent probes. Nat. Protoc. 6(7):991–1009, 2011.
Vocadlo, D. J., H. C. Hang, et al. A chemical approach for identifying O-GlcNAc-modified proteins in cells. Proc. Natl. Acad. Sci. USA 100(16):9116–9121, 2003.
Vosseller, K., L. Wells, et al. Elevated nucleocytoplasmic glycosylation by O-GlcNAc results in insulin resistance associated with defects in Akt activation in 3T3-L1 adipocytes. Proc. Natl. Acad. Sci. USA 99(8):5313–5318, 2002.
Wang, S., X. Huang, et al. Extensive crosstalk between O-GlcNAcylation and phosphorylation regulates Akt signaling. PLoS One 75:e37427, 2012.
Wang, J., M. Torii, et al. dbOGAP—an integrated bioinformatics resource for protein O-GlcNAcylation. BMC Bioinform. 12:91, 2011.
Wells, L., K. Vosseller, et al. Glycosylation of nucleocytoplasmic proteins: signal transduction and O-GlcNAc. Science 291(5512):2376–2378, 2001.
Zachara, N. E., N. O’Donnell, et al. Dynamic O-GlcNAc modification of nucleocytoplasmic proteins in response to stress. A survival response of mammalian cells. J. Biol. Chem. 279(29):30133–30142, 2004.
Zhu, Y., T. W. Liu, et al. Post-translational O-GlcNAcylation is essential for nuclear pore integrity and maintenance of the pore selectivity filter. J Mol. Cell Biol. 2015.
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The Project was supported by Grants from the National Institutes of Health (NIH GM094041, GM097037 and GM116204).
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Andrew Ruba and Weidong Yang declare that they have no conflicts of interest.
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Ruba, A., Yang, W. O-GlcNAc-ylation in the Nuclear Pore Complex. Cel. Mol. Bioeng. 9, 227–233 (2016). https://doi.org/10.1007/s12195-016-0440-0
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DOI: https://doi.org/10.1007/s12195-016-0440-0