Lutzmann, Malik and Kunze, Ruth and Stangl, Karin and Stelter, Philipp and Fejes Tóth, Katalin and Böttcher, Bettina and Hurt, Ed (2005) Reconstitution of Nup157 and Nup145N into the Nup84 Complex. Journal of Biological Chemistry, 280 (18). pp. 18442-18451. ISSN 0021-9258. doi:10.1074/jbc.m412787200. https://resolver.caltech.edu/CaltechAUTHORS:20200421-090412153
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Abstract
About 30 different nucleoporins (Nups) constitute the nuclear pore complex. We have affinity-purified 28 of these nuclear pore proteins and identified new nucleoporin interactions by this analysis. We found that Nup157 and Nup170, two members of the large structural Nups, and the Gly-Leu-Phe-Gly nucleoporin Nup145N specifically co-purified with members of the Nup84 complex. In addition, Nup145N co-enriched during Nup157 purification. By in vitro reconstitution, we demonstrate that Nup157 and Nup145N form a nucleoporin subcomplex. Moreover, we show that Nup157 and Nup145N bind to the heptameric Nup84 complex. This assembly thus represents approximately one-third of all nucleoporins. To characterize Nup157 structurally, we purified and analyzed it by electron microscopy. Nup157 is a hollow sphere that resembles a clamp or a gripping hand. Thus, we could reconstitute an interaction between a large structural Nup, an FG repeat Nup, and a major structural module of the nuclear pore complex.
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| Additional Information: | © 2005 The American Society for Biochemistry and Molecular Biology, Inc. Received November 11, 2004. Revision received January 28, 2005. We are grateful to Daniela Roser for technical assistance in TAP-tagging the yeast nucleoporins, Dr. Katja Strässer for the Nup60-TAP purification, Denise Lau for the high copy number ProtA-Nup192 plasmid, Sabine Merker and Petra Ihrig, under the supervision of Dr. J. Lechner (BZH, Heidelberg, Germany), for performing mass spectrometry, and Karsten Rippe (Kirchhoff-Institut für Physik, Heidelberg University) for help in analytical ultracentrifugation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. | |||||||||
| Issue or Number: | 18 | |||||||||
| DOI: | 10.1074/jbc.m412787200 | |||||||||
| Record Number: | CaltechAUTHORS:20200421-090412153 | |||||||||
| Persistent URL: | https://resolver.caltech.edu/CaltechAUTHORS:20200421-090412153 | |||||||||
| Usage Policy: | No commercial reproduction, distribution, display or performance rights in this work are provided. | |||||||||
| ID Code: | 102690 | |||||||||
| Collection: | CaltechAUTHORS | |||||||||
| Deposited By: | Tony Diaz | |||||||||
| Deposited On: | 21 Apr 2020 16:11 | |||||||||
| Last Modified: | 16 Nov 2021 18:14 |
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