Abstract
Nucleocytoplasmic transport occurs through nuclear pore complexes (NPCs) embedded in the nuclear envelope1. Here, we discovered an unexpected role for yeast dynein light chain (Dyn2)2 in the NPC. Dyn2 is a previously undescribed nucleoporin that functions as molecular glue to dimerize and stabilize the Nup82–Nsp1–Nup159 complex, a module of the cytoplasmic pore filaments3. Biochemical analyses showed that Dyn2 binds to a linear motif (termed DIDNup159) inserted between the Phe-Gly repeat and coiled-coil domain of Nup159. Electron microscopy revealed that the reconstituted Dyn2–DIDNup159 complex forms a rigid rod-like structure, in which five Dyn2 homodimers align like 'pearls on a string' between two extented DIDNup159 strands. These findings imply that the rigid 20 nm long Dyn2–DIDNup159 filament projects the Nup159 Phe-Gly repeats from the Nup82 module. Thus, it is possible that dynein light chain plays a role in organizing natively unfolded Phe-Gly repeats within the NPC scaffold to facilitate nucleocytoplasmic transport.
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Acknowledgements
We are grateful to S. Merker, P. Ihrig and J. Lechner for performing mass spectrometry. E.H. is recipient of grants from the Deutsche Forschungsgemeinschaft (SFB 638/B2) and Fonds der Chemischen Industrie.
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Experiments were designed and data analysed and interpreted by P.S. and E.H. Strain construction, DNA recombination work, fluorescence microscopy and biochemical analyses (affinity purification, gel filtration, in vitro assays) were performed by P.S. and R.K. D.H. and P.P. performed double-flourescence microscopy. Negative-staining electron microscopy was conducted by D.F., M.D. and B.M. The manuscript was written by P.S. and E.H. All authors discussed the results and commented on the manuscript.
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Supplementary figures S1, S2, S3, S4, S5, S6 and Supplementary table S1 (PDF 1204 kb)
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Stelter, P., Kunze, R., Flemming, D. et al. Molecular basis for the functional interaction of dynein light chain with the nuclear-pore complex. Nat Cell Biol 9, 788–796 (2007). https://doi.org/10.1038/ncb1604
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DOI: https://doi.org/10.1038/ncb1604
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