| Autor: |
García-Santisteban, Iraia, Arregi, Igor, Alonso-Mariño, Marián, Urbaneja, María, Garcia-Vallejo, Juan, Bañuelos, Sonia, Rodríguez, Jose |
| Předmět: |
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| Zdroj: |
Cellular & Molecular Life Sciences; Dec2016, Vol. 73 Issue 24, p4685-4699, 15p, 1 Diagram, 6 Graphs |
| Abstrakt: |
The exportin CRM1 binds nuclear export signals (NESs), and mediates active transport of NES-bearing proteins from the nucleus to the cytoplasm. Structural and biochemical analyses have uncovered the molecular mechanisms underlying CRM1/NES interaction. CRM1 binds NESs through a hydrophobic cleft, whose open or closed conformation facilitates NES binding and release. Several cofactors allosterically modulate the conformation of the NES-binding cleft through intramolecular interactions involving an acidic loop and a C-terminal helix in CRM1. This current model of CRM1-mediated nuclear export has not yet been evaluated in a cellular setting. Here, we describe SRV100, a cellular reporter to interrogate CRM1 nuclear export activity. Using this novel tool, we provide evidence further validating the model of NES binding and release by CRM1. Furthermore, using both SRV100-based cellular assays and in vitro biochemical analyses, we investigate the functional consequences of a recurrent cancer-related mutation, which targets a residue near CRM1 NES-binding cleft. Our data indicate that this mutation does not necessarily abrogate the nuclear export activity of CRM1, but may increase its affinity for NES sequences bearing a more negatively charged C-terminal end. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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