Abstract
The ubiquitin-like protein SUMO-1 (small ubiquitin-related modifier 1) is covalently attached to substrate proteins by ligases and cleaved by isopeptidases. Yeast has two SUMO-1-deconjugating enzymes, Ulp1 and Ulp2, which are located at nuclear pores and in the nucleoplasm, respectively. Here we show that the catalytic C-domain of Ulp1 must be excluded from the nucleoplasm for cell viability. This is achieved by the noncatalytic N-domain, which tethers Ulp1 to the nuclear pores. The bulk of cellular Ulp1 is not associated with nucleoporins but instead associates with three karyopherins (Pse1, Kap95 and Kap60), in a complex that is not dissociated by RanGTP in vitro. The Ulp1 N-domain has two distinct binding sites for Pse1 and Kap95/Kap60, both of which are required for anchoring to the nuclear pore complex. We propose that Ulp1 is tethered to the nuclear pores by a Ran-insensitive interaction with karyopherins associated with nucleoporins. This location could allow Ulp1 to remove SUMO-1 from sumoylated cargo proteins during their passage through the nuclear pore channel.
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Acknowledgements
E.H. was supported by grants from the Deutsche Forschungsgemeinschaft (Leibniz-Programm) and Fonds der Chemischen Industrie. V.G.P. is the recipient of a long-term fellowship from the Human Frontier Science Program.
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Supplementary Tables
Table 1: Yeast strains (DOC 28 kb)
Table 2: Plasmids and constructions
Legend to supplementary Figure 1.
Supplementary Figure
Fig. 1. Levels of Ulp1-GFP are not altered upon ovexpression of Yrb4ΔN or in the pse1-1 strain when shifted to restrictive temperature. (JPG 391 kb)
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Panse, V., Küster, B., Gerstberger, T. et al. Unconventional tethering of Ulp1 to the transport channel of the nuclear pore complex by karyopherins. Nat Cell Biol 5, 21–27 (2003). https://doi.org/10.1038/ncb893
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DOI: https://doi.org/10.1038/ncb893
