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Enhanced dimerization drives ligand-independent activity of mutant epidermal growth factor receptor in lung cancer

Mutations within the epidermal growth factor receptor (EGFR/erbB1/Her1) are often associated with tumorigenesis. In particular, a number of EGFR mutants that demonstrate ligand-independent signaling are common in non–small cell lung cancer (NSCLC), including kinase domain mutations L858R (also calle...

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Autores principales: Valley, Christopher C., Arndt-Jovin, Donna J., Karedla, Narain, Steinkamp, Mara P., Chizhik, Alexey I., Hlavacek, William S., Wilson, Bridget S., Lidke, Keith A., Lidke, Diane S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4710239/
https://www.ncbi.nlm.nih.gov/pubmed/26337388
http://dx.doi.org/10.1091/mbc.E15-05-0269
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author Valley, Christopher C.
Arndt-Jovin, Donna J.
Karedla, Narain
Steinkamp, Mara P.
Chizhik, Alexey I.
Hlavacek, William S.
Wilson, Bridget S.
Lidke, Keith A.
Lidke, Diane S.
author_facet Valley, Christopher C.
Arndt-Jovin, Donna J.
Karedla, Narain
Steinkamp, Mara P.
Chizhik, Alexey I.
Hlavacek, William S.
Wilson, Bridget S.
Lidke, Keith A.
Lidke, Diane S.
author_sort Valley, Christopher C.
collection PubMed
description Mutations within the epidermal growth factor receptor (EGFR/erbB1/Her1) are often associated with tumorigenesis. In particular, a number of EGFR mutants that demonstrate ligand-independent signaling are common in non–small cell lung cancer (NSCLC), including kinase domain mutations L858R (also called L834R) and exon 19 deletions (e.g., ΔL747-P753insS), which collectively make up nearly 90% of mutations in NSCLC. The molecular mechanisms by which these mutations confer constitutive activity remain unresolved. Using multiple subdiffraction-limit imaging modalities, we reveal the altered receptor structure and interaction kinetics of NSCLC-associated EGFR mutants. We applied two-color single quantum dot tracking to quantify receptor dimerization kinetics on living cells and show that, in contrast to wild-type EGFR, mutants are capable of forming stable, ligand-independent dimers. Two-color superresolution localization microscopy confirmed ligand-independent aggregation of EGFR mutants. Live-cell Förster resonance energy transfer measurements revealed that the L858R kinase mutation alters ectodomain structure such that unliganded mutant EGFR adopts an extended, dimerization-competent conformation. Finally, mutation of the putative dimerization arm confirmed a critical role for ectodomain engagement in ligand-independent signaling. These data support a model in which dysregulated activity of NSCLC-associated kinase mutants is driven by coordinated interactions involving both the kinase and extracellular domains that lead to enhanced dimerization.
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spelling pubmed-47102392016-01-20 Enhanced dimerization drives ligand-independent activity of mutant epidermal growth factor receptor in lung cancer Valley, Christopher C. Arndt-Jovin, Donna J. Karedla, Narain Steinkamp, Mara P. Chizhik, Alexey I. Hlavacek, William S. Wilson, Bridget S. Lidke, Keith A. Lidke, Diane S. Mol Biol Cell Articles Mutations within the epidermal growth factor receptor (EGFR/erbB1/Her1) are often associated with tumorigenesis. In particular, a number of EGFR mutants that demonstrate ligand-independent signaling are common in non–small cell lung cancer (NSCLC), including kinase domain mutations L858R (also called L834R) and exon 19 deletions (e.g., ΔL747-P753insS), which collectively make up nearly 90% of mutations in NSCLC. The molecular mechanisms by which these mutations confer constitutive activity remain unresolved. Using multiple subdiffraction-limit imaging modalities, we reveal the altered receptor structure and interaction kinetics of NSCLC-associated EGFR mutants. We applied two-color single quantum dot tracking to quantify receptor dimerization kinetics on living cells and show that, in contrast to wild-type EGFR, mutants are capable of forming stable, ligand-independent dimers. Two-color superresolution localization microscopy confirmed ligand-independent aggregation of EGFR mutants. Live-cell Förster resonance energy transfer measurements revealed that the L858R kinase mutation alters ectodomain structure such that unliganded mutant EGFR adopts an extended, dimerization-competent conformation. Finally, mutation of the putative dimerization arm confirmed a critical role for ectodomain engagement in ligand-independent signaling. These data support a model in which dysregulated activity of NSCLC-associated kinase mutants is driven by coordinated interactions involving both the kinase and extracellular domains that lead to enhanced dimerization. The American Society for Cell Biology 2015-11-05 /pmc/articles/PMC4710239/ /pubmed/26337388 http://dx.doi.org/10.1091/mbc.E15-05-0269 Text en © 2015 Valley et al. 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 (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.
spellingShingle Articles
Valley, Christopher C.
Arndt-Jovin, Donna J.
Karedla, Narain
Steinkamp, Mara P.
Chizhik, Alexey I.
Hlavacek, William S.
Wilson, Bridget S.
Lidke, Keith A.
Lidke, Diane S.
Enhanced dimerization drives ligand-independent activity of mutant epidermal growth factor receptor in lung cancer
title Enhanced dimerization drives ligand-independent activity of mutant epidermal growth factor receptor in lung cancer
title_full Enhanced dimerization drives ligand-independent activity of mutant epidermal growth factor receptor in lung cancer
title_fullStr Enhanced dimerization drives ligand-independent activity of mutant epidermal growth factor receptor in lung cancer
title_full_unstemmed Enhanced dimerization drives ligand-independent activity of mutant epidermal growth factor receptor in lung cancer
title_short Enhanced dimerization drives ligand-independent activity of mutant epidermal growth factor receptor in lung cancer
title_sort enhanced dimerization drives ligand-independent activity of mutant epidermal growth factor receptor in lung cancer
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4710239/
https://www.ncbi.nlm.nih.gov/pubmed/26337388
http://dx.doi.org/10.1091/mbc.E15-05-0269
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