Unknown

Dataset Information

0

Probing nuclear pore complex architecture with proximity-dependent biotinylation.


ABSTRACT: Proximity-dependent biotin identification (BioID) is a method for identifying protein associations that occur in vivo. By fusing a promiscuous biotin ligase to a protein of interest expressed in living cells, BioID permits the labeling of proximate proteins during a defined labeling period. In this study we used BioID to study the human nuclear pore complex (NPC), one of the largest macromolecular assemblies in eukaryotes. Anchored within the nuclear envelope, NPCs mediate the nucleocytoplasmic trafficking of numerous cellular components. We applied BioID to constituents of the Nup107-160 complex and the Nup93 complex, two conserved NPC subcomplexes. A strikingly different set of NPC constituents was detected depending on the position of these BioID-fusion proteins within the NPC. By applying BioID to several constituents located throughout the extremely stable Nup107-160 subcomplex, we refined our understanding of this highly conserved subcomplex, in part by demonstrating a direct interaction of Nup43 with Nup85. Furthermore, by using the extremely stable Nup107-160 structure as a molecular ruler, we defined the practical labeling radius of BioID. These studies further our understanding of human NPC organization and demonstrate that BioID is a valuable tool for exploring the constituency and organization of large protein assemblies in living cells.

SUBMITTER: Kim DI 

PROVIDER: S-EPMC4066523 | biostudies-literature | 2014 Jun

REPOSITORIES: biostudies-literature

altmetric image

Publications

Probing nuclear pore complex architecture with proximity-dependent biotinylation.

Kim Dae In DI   Birendra K C KC   Zhu Wenhong W   Motamedchaboki Khatereh K   Doye Valérie V   Roux Kyle J KJ  

Proceedings of the National Academy of Sciences of the United States of America 20140603 24


Proximity-dependent biotin identification (BioID) is a method for identifying protein associations that occur in vivo. By fusing a promiscuous biotin ligase to a protein of interest expressed in living cells, BioID permits the labeling of proximate proteins during a defined labeling period. In this study we used BioID to study the human nuclear pore complex (NPC), one of the largest macromolecular assemblies in eukaryotes. Anchored within the nuclear envelope, NPCs mediate the nucleocytoplasmic  ...[more]

Similar Datasets

| S-EPMC8890125 | biostudies-literature
| S-EPMC5180592 | biostudies-literature
| S-EPMC4774922 | biostudies-literature
| S-EPMC8550940 | biostudies-literature
| S-EPMC4826903 | biostudies-literature
| S-EPMC6006428 | biostudies-literature
| S-EPMC3431207 | biostudies-literature
| S-EPMC1413685 | biostudies-literature
2023-12-04 | PXD047268 | Pride
| S-EPMC2865169 | biostudies-literature