J. Cell Biol., Volume 140, Number 2, January 26, 1998 259-270

Molecular Characterization of the SUMO-1 Modification of RanGAP1 and Its Role in Nuclear Envelope Association

Rohit Mahajan, Larry Gerace, and Frauke Melchior

Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037

The mammalian guanosine triphosphate (GTP)ase-activating protein RanGAP1 is the first example of a protein covalently linked to the ubiquitin-related protein SUMO-1. Here we used peptide mapping, mass spectroscopy analysis, and mutagenesis to identify the nature of the link between RanGAP1 and SUMO-1. SUMO-1 is linked to RanGAP1 via glycine 97, indicating that the last 4 amino acids of this 101- amino acid protein are proteolytically removed before its attachment to RanGAP1. Recombinant SUMO-1 lacking the last four amino acids is efficiently used for modification of RanGAP1 in vitro and of multiple unknown proteins in vivo. In contrast to most ubiquitinated proteins, only a single lysine residue (K526) in RanGAP1 can serve as the acceptor site for modification by SUMO-1. Modification of RanGAP1 with SUMO-1 leads to association of RanGAP1 with the nuclear envelope (NE), where it was previously shown to be required for nuclear protein import. Sufficient information for modification and targeting resides in a 25-kD domain of RanGAP1. RanGAP1-SUMO-1 remains stably associated with the NE during many cycles of in vitro import. This indicates that removal of RanGAP1 from the NE is not a required element of nuclear protein import and suggests that the reversible modification of RanGAP1 may have a regulatory role.

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• Kunzler, M., Hurt, E. (2002). Targeting of Ran: variation on a common theme?. J. Cell Sci. 114: 3233-3241 [Abstract] [Full Text]  
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• Saitoh, H., Pizzi, M. D., Wang, J. (2002). Perturbation of SUMOlation Enzyme Ubc9 by Distinct Domain within Nucleoporin RanBP2/Nup358. J. Biol. Chem. 277: 4755-4763 [Abstract] [Full Text]  
• Nishida, T., Kaneko, F., Kitagawa, M., Yasuda, H. (2001). Characterization of a Novel Mammalian SUMO-1/Smt3-specific Isopeptidase, a Homologue of Rat Axam, Which Is an Axin-binding Protein Promoting beta -Catenin Degradation. J. Biol. Chem. 276: 39060-39066 [Abstract] [Full Text]  
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• Poukka, H., Karvonen, U., Janne, O. A., Palvimo, J. J. (2000). Covalent modification of the androgen receptor by small ubiquitin-like modifier 1 (SUMO-1). Proc. Natl. Acad. Sci. USA 97: 14145-14150 [Abstract] [Full Text]  
• LIU, L. F., DESAI, S. D., LI, T.-K., MAO, Y., SUN, M., SIM, S.-P. (2000). Mechanism of Action of Camptothecin. Ann. N. Y. Acad. Sci. 922: 1-10 [Abstract] [Full Text]  
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• Saitoh, H., Hinchey, J. (2000). Functional Heterogeneity of Small Ubiquitin-related Protein Modifiers SUMO-1 versus SUMO-2/3. J. Biol. Chem. 275: 6252-6258 [Abstract] [Full Text]  
• Gong, L., Millas, S., Maul, G. G., Yeh, E. T. H. (2000). Differential Regulation of Sentrinized Proteins by a Novel Sentrin-specific Protease. J. Biol. Chem. 275: 3355-3359 [Abstract] [Full Text]  
• Lehembre, F., Badenhorst, P., Müller, S., Travers, A., Schweisguth, F., Dejean, A. (2000). Covalent Modification of the Transcriptional Repressor Tramtrack by the Ubiquitin-Related Protein Smt3 in Drosophila Flies. Mol. Cell. Biol. 20: 1072-1082 [Abstract] [Full Text]  
• Singh, B. B., Patel, H. H., Roepman, R., Schick, D., Ferreira, P. A. (1999). The Zinc Finger Cluster Domain of RanBP2 Is a Specific Docking Site for the Nuclear Export Factor, Exportin-1. J. Biol. Chem. 274: 37370-37378 [Abstract] [Full Text]  
• Tanaka, K., Nishide, J., Okazaki, K., Kato, H., Niwa, O., Nakagawa, T., Matsuda, H., Kawamukai, M., Murakami, Y. (1999). Characterization of a Fission Yeast SUMO-1 Homologue, Pmt3p, Required for Multiple Nuclear Events, Including the Control of Telomere Length and Chromosome Segregation. Mol. Cell. Biol. 19: 8660-8672 [Abstract] [Full Text]  
• Johnson, E. S., Blobel, G. (1999). Cell Cycle-Regulated Attachment of the Ubiquitin-Related Protein SUMO to the Yeast Septins. J. Cell Biol. 147: 981-994 [Abstract] [Full Text]  
• Kim, Y. H., Choi, C. Y., Kim, Y. (1999). Covalent modification of the homeodomain-interacting protein kinase 2 (HIPK2) by the ubiquitin-like protein SUMO-1. Proc. Natl. Acad. Sci. USA 96: 12350-12355 [Abstract] [Full Text]  
• Yaseen, N. R., Blobel, G. (1999). GTP Hydrolysis Links Initiation and Termination of Nuclear Import on the Nucleoporin Nup358. J. Biol. Chem. 274: 26493-26502 [Abstract] [Full Text]  
• Poukka, H., Aarnisalo, P., Karvonen, U., Palvimo, J. J., Janne, O. A. (1999). Ubc9 Interacts with the Androgen Receptor and Activates Receptor-dependent Transcription. J. Biol. Chem. 274: 19441-19446 [Abstract] [Full Text]  
• Sternsdorf, T., Puccetti, E., Jensen, K., Hoelzer, D., Will, H., Ottmann, O. G., Ruthardt, M. (1999). PIC-1/SUMO-1-Modified PML-Retinoic Acid Receptor alpha  Mediates Arsenic Trioxide-Induced Apoptosis in Acute Promyelocytic Leukemia. Mol. Cell. Biol. 19: 5170-5178 [Abstract] [Full Text]  
• Yaseen, N. R., Blobel, G. (1999). Two distinct classes of Ran-binding sites on the nucleoporin Nup-358. Proc. Natl. Acad. Sci. USA 96: 5516-5521 [Abstract] [Full Text]  
• Yuan, W., Strømhaug, P. E., Dunn, W. A. Jr. (1999). Glucose-induced Autophagy of Peroxisomes in Pichia pastoris Requires a Unique E1-like Protein. Mol. Biol. Cell 10: 1353-1366 [Abstract] [Full Text]  
• Tanida, I., Mizushima, N., Kiyooka, M., Ohsumi, M., Ueno, T., Ohsumi, Y., Kominami, E. (1999). Apg7p/Cvt2p: A Novel Protein-activating Enzyme Essential for Autophagy. Mol. Biol. Cell 10: 1367-1379 [Abstract] [Full Text]  
• Sternsdorf, T., Jensen, K., Reich, B., Will, H. (1999). The Nuclear Dot Protein Sp100, Characterization of Domains Necessary for Dimerization, Subcellular Localization, and Modification by Small Ubiquitin-like Modifiers. J. Biol. Chem. 274: 12555-12566 [Abstract] [Full Text]  
• Merrill, C., Bayraktaroglu, L., Kusano, A., Ganetzky, B. (1999). Truncated RanGAP Encoded by the Segregation Distorter Locus of Drosophila. Science 283: 1742-1745 [Abstract] [Full Text]  
• Duprez, E, Saurin, A., Desterro, J., Lallemand-Breitenbach, V, Howe, K, Boddy, M., Solomon, E, de The, H, Hay, R., Freemont, P. (1999). SUMO-1 modification of the acute promyelocytic leukaemia protein PML: implications for nuclear localisation. J. Cell Sci. 112: 381-393 [Abstract]  
• Kamitani, T., Kito, K., Nguyen, H. P., Wada, H., Fukuda-Kamitani, T., Yeh, E. T. H. (1998). Identification of Three Major Sentrinization Sites in PML. J. Biol. Chem. 273: 26675-26682 [Abstract] [Full Text]  
• Ferreira, P. A., Yunfei, C., Schick, D., Roepman, R. (1998). The Cyclophilin-like Domain Mediates the Association of Ran-Binding Protein 2 with Subunits of the 19 S Regulatory Complex of the Proteasome. J. Biol. Chem. 273: 24676-24682 [Abstract] [Full Text]  
• Lamond, A. I., Earnshaw, W. C. (1998). Structure and Function in the Nucleus. Science 280: 547-553 [Abstract] [Full Text]  
• Mao, Y., Desai, S. D., Liu, L. F. (2000). SUMO-1 Conjugation to Human DNA Topoisomerase II Isozymes. J. Biol. Chem. 275: 26066-26073 [Abstract] [Full Text]  
• Rangasamy, D., Wilson, V. G. (2000). Bovine Papillomavirus E1 Protein Is Sumoylated by the Host Cell Ubc9 Protein. J. Biol. Chem. 275: 30487-30495 [Abstract] [Full Text]  
• Rangasamy, D., Woytek, K., Khan, S. A., Wilson, V. G. (2000). SUMO-1 Modification of Bovine Papillomavirus E1 Protein Is Required for Intranuclear Accumulation. J. Biol. Chem. 275: 37999-38004 [Abstract] [Full Text]  
• Goodson, M. L., Hong, Y., Rogers, R., Matunis, M. J., Park-Sarge, O.-K., Sarge, K. D. (2001). SUMO-1 Modification Regulates the DNA Binding Activity of Heat Shock Transcription Factor 2, a Promyelocytic Leukemia Nuclear Body Associated Transcription Factor. J. Biol. Chem. 276: 18513-18518 [Abstract] [Full Text]  
• Sampson, D. A., Wang, M., Matunis, M. J. (2001). The Small Ubiquitin-like Modifier-1 (SUMO-1) Consensus Sequence Mediates Ubc9 Binding and Is Essential for SUMO-1 Modification. J. Biol. Chem. 276: 21664-21669 [Abstract] [Full Text]  
• Mao, Y., Sun, M., Desai, S. D., Liu, L. F. (2000). SUMO-1 conjugation to topoisomerase I: A possible repair response to topoisomerase-mediated DNA damage. Proc. Natl. Acad. Sci. USA 97: 4046-4051 [Abstract] [Full Text]  
• Kurtzman, A. L., Schechter, N. (2001). Ubc9 interacts with a nuclear localization signal and mediates nuclear localization of the paired-like homeobox protein Vsx-1 independent of SUMO-1 modification. Proc. Natl. Acad. Sci. USA 98: 5602-5607 [Abstract] [Full Text]  
• Joseph, J., Tan, S.-H., Karpova, T. S., McNally, J. G., Dasso, M. (2002). SUMO-1 targets RanGAP1 to kinetochores and mitotic spindles. J. Cell Biol. 156: 595-602 [Abstract] [Full Text]  

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