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Recognition of nuclear export signals by CRM1 carrying the oncogenic E571K mutation

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...

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Autores principales: Baumhardt, Jordan M., Walker, Janek S., Lee, Yoonji, Shakya, Binita, Brautigam, Chad A., Lapalombella, Rosa, Grishin, Nick, Chook, Yuh Min
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7525811/
https://www.ncbi.nlm.nih.gov/pubmed/32520643
http://dx.doi.org/10.1091/mbc.E20-04-0233
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author Baumhardt, Jordan M.
Walker, Janek S.
Lee, Yoonji
Shakya, Binita
Brautigam, Chad A.
Lapalombella, Rosa
Grishin, Nick
Chook, Yuh Min
author_facet Baumhardt, Jordan M.
Walker, Janek S.
Lee, Yoonji
Shakya, Binita
Brautigam, Chad A.
Lapalombella, Rosa
Grishin, Nick
Chook, Yuh Min
author_sort Baumhardt, Jordan M.
collection PubMed
description 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.
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spelling pubmed-75258112020-10-16 Recognition of nuclear export signals by CRM1 carrying the oncogenic E571K mutation Baumhardt, Jordan M. Walker, Janek S. Lee, Yoonji Shakya, Binita Brautigam, Chad A. Lapalombella, Rosa Grishin, Nick Chook, Yuh Min Mol Biol Cell Articles 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. The American Society for Cell Biology 2020-08-01 /pmc/articles/PMC7525811/ /pubmed/32520643 http://dx.doi.org/10.1091/mbc.E20-04-0233 Text en © 2020 Baumhardt et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0 This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License.
spellingShingle Articles
Baumhardt, Jordan M.
Walker, Janek S.
Lee, Yoonji
Shakya, Binita
Brautigam, Chad A.
Lapalombella, Rosa
Grishin, Nick
Chook, Yuh Min
Recognition of nuclear export signals by CRM1 carrying the oncogenic E571K mutation
title Recognition of nuclear export signals by CRM1 carrying the oncogenic E571K mutation
title_full Recognition of nuclear export signals by CRM1 carrying the oncogenic E571K mutation
title_fullStr Recognition of nuclear export signals by CRM1 carrying the oncogenic E571K mutation
title_full_unstemmed Recognition of nuclear export signals by CRM1 carrying the oncogenic E571K mutation
title_short Recognition of nuclear export signals by CRM1 carrying the oncogenic E571K mutation
title_sort recognition of nuclear export signals by crm1 carrying the oncogenic e571k mutation
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7525811/
https://www.ncbi.nlm.nih.gov/pubmed/32520643
http://dx.doi.org/10.1091/mbc.E20-04-0233
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