Analysis of the initiation of nuclear pore assembly by ectopically targeting nucleoporins to chromatin
| Autor: | Anna Travesa, Michal Schwartz, Steven W Martell, Douglass J. Forbes |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Nup133
Nuclear Envelope nuclear pore assembly Nup160/107 complex Active Transport Cell Nucleus Lac repressor Biology medicine.disease_cause Sudden death 03 medical and health sciences cardiac arrhythmia 0302 clinical medicine LacI Nup155 R391H Cell Line Tumor medicine Lac Repressors Humans Nup153 Nuclear pore LacO array 030304 developmental biology 0303 health sciences Mutation Arrhythmias Cardiac Cell Biology Transfection Fusion protein Molecular biology Chromatin Cell biology Protein Structure Tertiary Nuclear Pore Complex Proteins nuclear rim localization ELYS Nucleoporin 030217 neurology & neurosurgery Research Paper |
| Zdroj: | Nucleus |
| ISSN: | 1949-1042 1949-1034 |
| Popis: | Nuclear pore complexes (NPCs) form the gateway to the nucleus, mediating virtually all nucleocytoplasmic trafficking. Assembly of a nuclear pore complex requires the organization of many soluble sub-complexes into a final massive structure embedded in the nuclear envelope. By use of a LacI/LacO reporter system, we were able to assess nucleoporin (Nup) interactions, show that they occur with a high level of specificity, and identify nucleoporins sufficient for initiation of the complex process of NPC assembly in vivo. Eleven nucleoporins from different sub-complexes were fused to LacI-CFP and transfected separately into a human cell line containing a stably integrated LacO DNA array. The LacI-Nup fusion proteins, which bound to the array, were examined for their ability to recruit endogenous nucleoporins to the intranuclear LacO site. Many could recruit nucleoporins of the same sub-complex and a number could also recruit other sub-complexes. Strikingly, Nup133 and Nup107 of the Nup107/160 subcomplex and Nup153 and Nup50 of the nuclear pore basket recruited a near full complement of nucleoporins to the LacO array. Furthermore, Nup133 and Nup153 efficiently targeted the LacO array to the nuclear periphery. Our data support a hierarchical, seeded assembly pathway and identify Nup133 and Nup153 as effective "seeds" for NPC assembly. In addition, we show that this system can be applied to functional studies of individual nucleoporin domains as well as to specific nucleoporin disease mutations. We find that the R391H cardiac arrhythmia/sudden death mutation of Nup155 prevents both its subcomplex assembly and nuclear rim targeting of the LacO array. |
| Databáze: | OpenAIRE |
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