Monoclonal antibodies identify a group of nuclear pore complex glycoproteins

doi:10.1083/jcb.104.5.1143

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Monoclonal antibodies identify a group of nuclear pore complex glycoproteins

CM Snow, A Senior and L Gerace

Using monoclonal antibodies we identified a group of eight polypeptides of rat liver nuclear envelopes that have common epitopes. Most or all of these proteins are structurally distinct, as shown by tryptic peptide mapping and analysis with polyclonal antibodies. While these polypeptides are relatively tightly bound to nuclear membranes, only one is an integral membrane protein. The eight antigens cofractionate with the nuclear pore complex under various conditions of ionic strength and detergent. It can be seen by immunofluorescence microscopy that the monoclonal antibodies reacting with these antigens stain the nuclear surface of interphase cells in a finely punctate pattern. When the nuclear envelope is disassembled and subsequently reformed during mitosis, the proteins are reversibly dispersed throughout the cytoplasm in the form of minute foci. By EM immunogold localization on isolated nuclear envelopes, the monoclonal antibodies label exclusively the nuclear pore complex, at both its nucleoplasmic and cytoplasmic margins. Considered together, our biochemical and localization data indicate that the eight nuclear envelope polypeptides are pore complex components. As shown in the accompanying paper (Holt, G. D., C. M. Snow, A. Senior, R. S. Haltiwanger, L. Gerace, and G. W. Hart, J. Cell Biol., 104:1157-1164) these eight polypeptides contain a novel form of glycosylation, O-linked N-acetylglucosamine. The relative abundance and disposition of these O-linked glycoproteins in the pore complex are consistent with their having a role in nucleocytoplasmic transport.
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Hofemeister, H., O'Hare, P. (2008). Nuclear Pore Composition and Gating in Herpes Simplex Virus-Infected Cells. J. Virol. 82: 8392-8399 [Abstract] [Full Text]
Katsani, K. R., Karess, R. E., Dostatni, N., Doye, V. (2008). In Vivo Dynamics of Drosophila Nuclear Envelope Components. Mol. Biol. Cell 19: 3652-3666 [Abstract] [Full Text]
Rengifo, J., Gibson, C. J., Winkler, E., Collin, T., Ehrlich, B. E. (2007). Regulation of the Inositol 1,4,5-Trisphosphate Receptor Type I by O-GlcNAc Glycosylation. J. Neurosci. 27: 13813-13821 [Abstract] [Full Text]
Naim, B., Brumfeld, V., Kapon, R., Kiss, V., Nevo, R., Reich, Z. (2007). Passive and Facilitated Transport in Nuclear Pore Complexes Is Largely Uncoupled. J. Biol. Chem. 282: 3881-3888 [Abstract] [Full Text]
Akimoto, Y., Hart, G. W., Wells, L., Vosseller, K., Yamamoto, K., Munetomo, E., Ohara-Imaizumi, M., Nishiwaki, C., Nagamatsu, S., Hirano, H., Kawakami, H. (2007). Elevation of the post-translational modification of proteins by O-linked N-acetylglucosamine leads to deterioration of the glucose-stimulated insulin secretion in the pancreas of diabetic Goto-Kakizaki rats. Glycobiology 17: 127-140 [Abstract] [Full Text]
Xylourgidis, N., Roth, P., Sabri, N., Tsarouhas, V., Samakovlis, C. (2006). The nucleoporin Nup214 sequesters CRM1 at the nuclear rim and modulates NF{kappa}B activation in Drosophila. J. Cell Sci. 119: 4409-4419 [Abstract] [Full Text]
Hutten, S., Kehlenbach, R. H. (2006). Nup214 Is Required for CRM1-Dependent Nuclear Protein Export In Vivo.. Mol. Cell. Biol. 26: 6772-6785 [Abstract] [Full Text]
Hu, P., Berkowitz, P., Madden, V. J., Rubenstein, D. S. (2006). Stabilization of Plakoglobin and Enhanced Keratinocyte Cell-Cell Adhesion by Intracellular O-Glycosylation. J. Biol. Chem. 281: 12786-12791 [Abstract] [Full Text]
Love, D. C., Hanover, J. A. (2005). The Hexosamine Signaling Pathway: Deciphering the "O-GlcNAc Code". Sci Signal 2005: re13-re13 [Abstract] [Full Text]
Onischenko, E. A., Gubanova, N. V., Kiseleva, E. V., Hallberg, E. (2005). Cdk1 and Okadaic Acid-sensitive Phosphatases Control Assembly of Nuclear Pore Complexes in Drosophila Embryos. Mol. Biol. Cell 16: 5152-5162 [Abstract] [Full Text]
Ludemann, N., Clement, A., Hans, V. H., Leschik, J., Behl, C., Brandt, R. (2005). O-Glycosylation of the Tail Domain of Neurofilament Protein M in Human Neurons and in Spinal Cord Tissue of a Rat Model of Amyotrophic Lateral Sclerosis (ALS). J. Biol. Chem. 280: 31648-31658 [Abstract] [Full Text]
Hanover, J. A., Forsythe, M. E., Hennessey, P. T., Brodigan, T. M., Love, D. C., Ashwell, G., Krause, M. (2005). A Caenorhabditis elegans model of insulin resistance: Altered macronutrient storage and dauer formation in an OGT-1 knockout. Proc. Natl. Acad. Sci. USA 102: 11266-11271 [Abstract] [Full Text]
Ohba, T., Schirmer, E. C., Nishimoto, T., Gerace, L. (2004). Energy- and temperature-dependent transport of integral proteins to the inner nuclear membrane via the nuclear pore. JCB 167: 1051-1062 [Abstract] [Full Text]
Liu, F., Iqbal, K., Grundke-Iqbal, I., Hart, G. W., Gong, C.-X. (2004). O-GlcNAcylation regulates phosphorylation of tau: A mechanism involved in Alzheimer's disease. Proc. Natl. Acad. Sci. USA 101: 10804-10809 [Abstract] [Full Text]
Hu, Y., Riesland, L., Paterson, A. J., Kudlow, J. E. (2004). Phosphorylation of Mouse Glutamine-Fructose-6-phosphate Amidotransferase 2 (GFAT2) by cAMP-dependent Protein Kinase Increases the Enzyme Activity. J. Biol. Chem. 279: 29988-29993 [Abstract] [Full Text]
Hazel, M., Cooksey, R. C., Jones, D., Parker, G., Neidigh, J. L., Witherbee, B., Gulve, E. A., McClain, D. A. (2004). Activation of the Hexosamine Signaling Pathway in Adipose Tissue Results in Decreased Serum Adiponectin and Skeletal Muscle Insulin Resistance. Endocrinology 145: 2118-2128 [Abstract] [Full Text]
Kang, H. T., Ju, J. W., Cho, J. W., Hwang, E. S. (2003). Down-regulation of Sp1 Activity through Modulation of O-Glycosylation by Treatment with a Low Glucose Mimetic, 2-Deoxyglucose. J. Biol. Chem. 278: 51223-51231 [Abstract] [Full Text]
Akimoto, Y., Kawakami, H., Yamamoto, K., Munetomo, E., Hida, T., Hirano, H. (2003). Elevated Expression of O-GlcNAc-Modified Proteins and O-GlcNAc Transferase in Corneas of Diabetic Goto-Kakizaki Rats. IOVS 44: 3802-3809 [Abstract] [Full Text]
Castagnet, P., Mavlyutov, T., Cai, Y., Zhong, F., Ferreira, P. (2003). RPGRIP1s with distinct neuronal localization and biochemical properties associate selectively with RanBP2 in amacrine neurons. Hum Mol Genet 12: 1847-1863 [Abstract] [Full Text]
Rawe, V.Y., Olmedo, S.B., Nodar, F.N., Ponzio, R., Sutovsky, P. (2003). Abnormal assembly of annulate lamellae and nuclear pore complexes coincides with fertilization arrest at the pronuclear stage of human zygotic development. Hum Reprod 18: 576-582 [Abstract] [Full Text]
McClain, D. A., Lubas, W. A., Cooksey, R. C., Hazel, M., Parker, G. J., Love, D. C., Hanover, J. A. (2002). Altered glycan-dependent signaling induces insulin resistance and hyperleptinemia. Proc. Natl. Acad. Sci. USA 99: 10695-10699 [Abstract] [Full Text]
Buse, M. G., Robinson, K. A., Marshall, B. A., Hresko, R. C., Mueckler, M. M. (2002). Enhanced O-GlcNAc protein modification is associated with insulin resistance in GLUT1-overexpressing muscles. Am. J. Physiol. Endocrinol. Metab. 283: E241-E250 [Abstract] [Full Text]
Juang, Y.-T., Solomou, E. E., Rellahan, B., Tsokos, G. C. (2002). Phosphorylation and O-Linked Glycosylation of Elf-1 Leads to Its Translocation to the Nucleus and Binding to the Promoter of the TCR {zeta}-Chain. J. Immunol. 168: 2865-2871 [Abstract] [Full Text]
Frosst, P., Guan, T., Subauste, C., Hahn, K., Gerace, L. (2002). Tpr is localized within the nuclear basket of the pore complex and has a role in nuclear protein export. JCB 156: 617-630 [Abstract] [Full Text]
Schmitt, I., Gerace, L. (2001). In Vitro Analysis of Nuclear Transport Mediated by the C-terminal Shuttle Domain of Tap. J. Biol. Chem. 276: 42355-42363 [Abstract] [Full Text]
HANOVER, J. A. (2001). Glycan-dependent signaling: O-linked N-acetylglucosamine. FASEB J. 15: 1865-1876 [Abstract] [Full Text]
Topper, L. M., Bastians, H., Ruderman, J. V., Gorbsky, G. J. (2001). Elevating the level of Cdc34/Ubc3 ubiquitin-conjugating enzyme in mitosis inhibits association of CENP-E with kinetochores and blocks the metaphase alignment of chromosomes. JCB 154: 707-718 [Abstract] [Full Text]
Tolonen, N., Doglio, L., Schleich, S., Locker, J. K. (2001). Vaccinia Virus DNA Replication Occurs in Endoplasmic Reticulum-enclosed Cytoplasmic Mini-Nuclei. Mol. Biol. Cell 12: 2031-2046 [Abstract] [Full Text]
Schirmer, E. C., Guan, T., Gerace, L. (2001). Involvement of the Lamin Rod Domain in Heterotypic Lamin Interactions Important for Nuclear Organization. JCB 153: 479-490 [Abstract] [Full Text]
Ben-Efraim, I., Gerace, L. (2001). Gradient of Increasing Affinity of Importin {beta} for Nucleoporins along the Pathway of Nuclear Import. JCB 152: 411-418 [Abstract] [Full Text]
Black, B. E., Holaska, J. M., Levesque, L., Ossareh-Nazari, B., Gwizdek, C., Dargemont, C., Paschal, B. M. (2001). Nxt1 Is Necessary for the Terminal Step of Crm1-Mediated Nuclear Export. JCB 152: 141-156 [Abstract] [Full Text]
Miller, B. R., Powers, M., Park, M., Fischer, W., Forbes, D. J. (2000). Identification of a New Vertebrate Nucleoporin, Nup188, with the Use of a Novel Organelle Trap Assay. Mol. Biol. Cell 11: 3381-3396 [Abstract] [Full Text]
Lee, K. K., Gruenbaum, Y., Spann, P., Liu, J., Wilson, K. L. (2000). C. elegans Nuclear Envelope Proteins Emerin, MAN1, Lamin, and Nucleoporins Reveal Unique Timing of Nuclear Envelope Breakdown during Mitosis. Mol. Biol. Cell 11: 3089-3099 [Abstract] [Full Text]
Guan, T., Kehlenbach, R. H., Schirmer, E. C., Kehlenbach, A., Fan, F., Clurman, B. E., Arnheim, N., Gerace, L. (2000). Nup50, a Nucleoplasmically Oriented Nucleoporin with a Role in Nuclear Protein Export. Mol. Cell. Biol. 20: 5619-5630 [Abstract] [Full Text]
Ojala, P. M., Sodeik, B., Ebersold, M. W., Kutay, U., Helenius, A. (2000). Herpes Simplex Virus Type 1 Entry into Host Cells: Reconstitution of Capsid Binding and Uncoating at the Nuclear Pore Complex In Vitro. Mol. Cell. Biol. 20: 4922-4931 [Abstract] [Full Text]
Wente, S. R. (2000). Gatekeepers of the Nucleus. Science 288: 1374-1377 [Abstract] [Full Text]
Liu, K., Paterson, A. J., Chin, E., Kudlow, J. E. (2000). Glucose stimulates protein modification by O-linked GlcNAc in pancreatic beta cells: Linkage of O-linked GlcNAc to beta cell death. Proc. Natl. Acad. Sci. USA 97: 2820-2825 [Abstract] [Full Text]
Steggerda, S. M., Black, B. E., Paschal, B. M. (2000). Monoclonal Antibodies to NTF2 Inhibit Nuclear Protein Import by Preventing Nuclear Translocation of the GTPase Ran. Mol. Biol. Cell 11: 703-719 [Abstract] [Full Text]
Black, B. E., Levesque, L., Holaska, J. M., Wood, T. C., Paschal, B. M. (1999). Identification of an NTF2-Related Factor That Binds Ran-GTP and Regulates Nuclear Protein Export. Mol. Cell. Biol. 19: 8616-8624 [Abstract] [Full Text]
Rolls, M. M., Stein, P. A., Taylor, S. S., Ha, E., McKeon, F., Rapoport, T. A. (1999). A Visual Screen of a Gfp-Fusion Library Identifies a New Type of Nuclear Envelope Membrane Protein. JCB 146: 29-44 [Abstract] [Full Text]
Kehlenbach, R. H., Dickmanns, A., Kehlenbach, A., Guan, T., Gerace, L. (1999). A Role for RanBP1 in the Release of CRM1 from the Nuclear Pore Complex in a Terminal Step of Nuclear Export. JCB 145: 645-657 [Abstract] [Full Text]
Rosorius, O, Reichart, B, Kratzer, F, Heger, P, Dabauvalle, M., Hauber, J (1999). Nuclear pore localization and nucleocytoplasmic transport of eIF-5A: evidence for direct interaction with the export receptor CRM1. J. Cell Sci. 112: 2369-2380 [Abstract]
Bangs, P., Burke, B., Powers, C., Craig, R., Purohit, A., Doxsey, S. (1998). Functional Analysis of Tpr: Identification of Nuclear Pore Complex Association and Nuclear Localization Domains and a Role in mRNA Export. JCB 143: 1801-1812 [Abstract] [Full Text]
de Hostos, E. L., McCaffrey, G., Sucgang, R., Pierce, D. W., Vale, R. D. (1998). A Developmentally Regulated Kinesin-related Motor Protein from Dictyostelium discoideum. Mol. Biol. Cell 9: 2093-2106 [Abstract] [Full Text]
Sutovsky, P, Simerly, C, Hewitson, L, Schatten, G (1998). Assembly of nuclear pore complexes and annulate lamellae promotes normal pronuclear development in fertilized mammalian oocytes. J. Cell Sci. 111: 2841-2854 [Abstract]
Yang, L., Guan, T., Gerace, L. (1997). Lamin-binding Fragment of LAP2 Inhibits Increase in Nuclear Volume during the Cell Cycle and Progression into S Phase. JCB 139: 1077-1087 [Abstract] [Full Text]
Delphin, C., Guan, T., Melchior, F., Gerace, L. (1997). RanGTP Targets p97 to RanBP2, a Filamentous Protein Localized at the Cytoplasmic Periphery of the Nuclear Pore Complex. Mol. Biol. Cell 8: 2379-2390 [Abstract] [Full Text]
Bastos, R., Pouplana, L. R. d., Enarson, M., Bodoor, K., Burke, B. (1997). Nup84, A Novel Nucleoporin That Is Associated With CAN/Nup214 on the Cytoplasmic Face of the Nuclear Pore Complex. JCB 137: 989-1000 [Abstract] [Full Text]
Del Priore, V, Heath, C, Snay, C, MacMillan, A, Gorsch, L, Dagher, S, Cole, C (1997). A structure/function analysis of Rat7p/Nup159p, an essential nucleoporin of Saccharomyces cerevisiae. J. Cell Sci. 110: 2987-2999 [Abstract]
Zimowska, G, Aris, J., Paddy, M. (1997). A Drosophila Tpr protein homolog is localized both in the extrachromosomal channel network and to nuclear pore complexes. J. Cell Sci. 110: 927-944 [Abstract]
Panté, N., Aebi, U. (1996). Sequential Binding of Import Ligands to Distinct Nucleopore Regions During Their Nuclear Import. Science 273: 1729-1732 [Abstract] [Full Text]
Sayeski, P. P., Kudlow, J. E. (1996). Glucose Metabolism to Glucosamine Is Necessary for Glucose Stimulation of Transforming Growth Factor-alpha Gene Transcription. J. Biol. Chem. 271: 15237-15243 [Abstract] [Full Text]
Weaver, V., Carson, C., Walker, P., Chaly, N, Lach, B, Raymond, Y, Brown, D., Sikorska, M (1996). Degradation of nuclear matrix and DNA cleavage in apoptotic thymocytes. J. Cell Sci. 109: 45-56 [Abstract]
Emig, S., Schmalz, D., Shakibaei, M., Buchner, K. (1995). The Nuclear Pore Complex Protein p62 Is One of Several Sialic Acid-containing Proteins of the Nuclear Envelope. J. Biol. Chem. 270: 13787-13793 [Abstract] [Full Text]
Martin, L., Crimaudo, C., Gerace, L. (1995). cDNA Cloning and Characterization of Lamina-associated Polypeptide 1C (LAP1C), an Integral Protein of the Inner Nuclear Membrane. J. Biol. Chem. 270: 8822-8828 [Abstract] [Full Text]
Blobel, G. (1995). Unidirectional and Bidirectional Protein Traffic across Membranes. Cold Spring Harb Symp Quant Biol 60: 1-10 [Abstract]
Melchior, F., Guan, T., Paschal, B., Gerace, L. (1995). Biochemical and Structural Analysis of Nuclear Protein Import. Cold Spring Harb Symp Quant Biol 60: 707-716 [Abstract]
Matsuoka, Y, Takechi, S, Nakayama, T, Yoneda, Y (1994). Exogenous histone H1 injection into mitotic cells disrupts synchronous progression of mitotic events by delaying chromosome decondensation. J. Cell Sci. 107: 693-701 [Abstract]
Buss, F, Kent, H, Stewart, M, Bailer, S., Hanover, J. (1994). Role of different domains in the self-association of rat nucleoporin p62. J. Cell Sci. 107: 631-638 [Abstract]
Chugani, D., Rome, L., Kedersha, N. (1993). Evidence that vault ribonucleoprotein particles localize to the nuclear pore complex. J. Cell Sci. 106: 23-29 [Abstract]
Goldberg, M., Allen, T. (1993). The nuclear pore complex: three-dimensional surface structure revealed by field emission, in-lens scanning electron microscopy, with underlying structure uncovered by proteolysis. J. Cell Sci. 106: 261-274 [Abstract]
Curtenaz, S., Peck, R. K. (1992). A monoclonal antibody study of protein distribution in the membrane skeleton of the ciliate Pseudomicrothorax. J. Cell Sci. 103: 1117-1125 [Abstract]
Dingwall, C, Laskey, R (1992). The nuclear membrane. Science 258: 942-947 [Abstract]
Yoneda, Y, Imamoto-Sonobe, N, Matsuoka, Y, Iwamoto, R, Kiho, Y, Uchida, T (1988). Antibodies to Asp-Asp-Glu-Asp can inhibit transport of nuclear proteins into the nucleus. Science 242: 275-278 [Abstract]
Chang, Q., Su, K., Baker, J. R., Yang, X., Paterson, A. J., Kudlow, J. E. (2000). Phosphorylation of Human Glutamine:Fructose-6-phosphate Amidotransferase by cAMP-dependent Protein Kinase at Serine 205 Blocks the Enzyme Activity. J. Biol. Chem. 275: 21981-21987 [Abstract] [Full Text]
Kehlenbach, R. H., Gerace, L. (2000). Phosphorylation of the Nuclear Transport Machinery Down-regulates Nuclear Protein Import in Vitro. J. Biol. Chem. 275: 17848-17856 [Abstract] [Full Text]
Kehlenbach, R. H., Assheuer, R., Kehlenbach, A., Becker, J., Gerace, L. (2001). Stimulation of Nuclear Export and Inhibition of Nuclear Import by a Ran Mutant Deficient in Binding to Ran-binding Protein 1. J. Biol. Chem. 276: 14524-14531 [Abstract] [Full Text]
Levesque, L., Guzik, B., Guan, T., Coyle, J., Black, B. E., Rekosh, D., Hammarskjold, M.-L., Paschal, B. M. (2001). RNA Export Mediated by Tap Involves NXT1-dependent Interactions with the Nuclear Pore Complex. J. Biol. Chem. 276: 44953-44962 [Abstract] [Full Text]
Frosst, P., Guan, T., Subauste, C., Hahn, K., Gerace, L. (2002). Tpr is localized within the nuclear basket of the pore complex and has a role in nuclear protein export. JCB 156: 617-630 [Abstract] [Full Text]
Brownawell, A. M., Macara, I. G. (2002). Exportin-5, a novel karyopherin, mediates nuclear export of double-stranded RNA binding proteins. JCB 156: 53-64 [Abstract] [Full Text]