6X2P

Crystal Structure of the Mek1NES peptide bound to CRM1

Summary for 6X2P

• Entry DOI: 10.2210/pdb6x2p/pdb
• Descriptor: GTP-binding nuclear protein Ran, Ran-specific GTPase-activating protein 1, Exportin-1, ... (8 entities in total)
• Functional Keywords: nuclear export, crm1, xpo1, exportin-1, protein transport
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 4
• Total formula weight: 161059.62

Authors

Baumhardt, J.M. (deposition date: 2020-05-20, release date: 2020-07-01, Last modification date: 2023-10-18)

Primary citation

Baumhardt, J.M.,Walker, J.S.,Lee, Y.,Shakya, B.,Brautigam, C.A.,Lapalombella, R.,Grishin, N.,Chook, Y.M.
Recognition of nuclear export signals by CRM1 carrying the oncogenic E571K mutation.
Mol.Biol.Cell, 31:1879-1891, 2020

PubMed Abstract: The E571K mutation of CRM1 is highly prevalent in some cancers, but its mechanism of tumorigenesis is unclear. Glu571 of CRM1 is located in its nuclear export signal (NES)-binding groove, suggesting that binding of select NESs may be altered. We generated HEK 293 cells with either monoallelic CRM1WT/E571K or biallelic CRM1E571K/E571K using CRISPR/Cas9. We also combined analysis of binding affinities and structures of 27 diverse NESs for wild-type and E571K CRM1 with structure-based bioinformatics. While most NESs bind the two CRM1 similarly, NESs from Mek1, eIF4E-transporter, and RPS2 showed >10-fold affinity differences. These NESs have multiple charged side chains binding close to CRM1 position 571, but this feature alone was not sufficient to predict different binding to CRM1(E571K). Consistent with eIF4E-transporter NES binding weaker to CRM1(E571K), eIF4E-transporter was mislocalized in tumor cells carrying CRM1(E571K). This serves as proof of concept that understanding how CRM1(E571K) affects NES binding provides a platform for identifying cargoes that are mislocalized in cancer upon CRM1 mutation. Finally, we showed that large affinity changes seen with some NES peptides (of Mek1 and RPS2) do not always translate to the full-length cargoes, suggesting limitations with current NES prediction methods. Therefore, comprehensive studies like ours are imperative to identify CRM1 cargoes with real pathogenic potential.

PubMed: 32520643
DOI: 10.1091/mbc.E20-04-0233

Experimental method

X-RAY DIFFRACTION (2.401 Å)