Cargando…

Conserved regulatory motifs in the juxtamembrane domain and kinase N-lobe revealed through deep mutational scanning of the MET receptor tyrosine kinase domain

MET is a receptor tyrosine kinase (RTK) responsible for initiating signaling pathways involved in development and wound repair. MET activation relies on ligand binding to the extracellular receptor, which prompts dimerization, intracellular phosphorylation, and recruitment of associated signaling pr...

Descripción completa

Detalles Bibliográficos
Autores principales: Estevam, Gabriella O., Linossi, Edmond M., Macdonald, Christian B., Espinoza, Carla A., Michaud, Jennifer M., Coyote-Maestas, Willow, Collisson, Eric A., Jura, Natalia, Fraser, James S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10418267/
https://www.ncbi.nlm.nih.gov/pubmed/37577651
http://dx.doi.org/10.1101/2023.08.03.551866
_version_ 1785088225994342400
author Estevam, Gabriella O.
Linossi, Edmond M.
Macdonald, Christian B.
Espinoza, Carla A.
Michaud, Jennifer M.
Coyote-Maestas, Willow
Collisson, Eric A.
Jura, Natalia
Fraser, James S.
author_facet Estevam, Gabriella O.
Linossi, Edmond M.
Macdonald, Christian B.
Espinoza, Carla A.
Michaud, Jennifer M.
Coyote-Maestas, Willow
Collisson, Eric A.
Jura, Natalia
Fraser, James S.
author_sort Estevam, Gabriella O.
collection PubMed
description MET is a receptor tyrosine kinase (RTK) responsible for initiating signaling pathways involved in development and wound repair. MET activation relies on ligand binding to the extracellular receptor, which prompts dimerization, intracellular phosphorylation, and recruitment of associated signaling proteins. Mutations, which are predominantly observed clinically in the intracellular juxtamembrane and kinase domains, can disrupt typical MET regulatory mechanisms. Understanding how juxtamembrane variants, such as exon 14 skipping (METΔEx14), and rare kinase domain mutations can increase signaling, often leading to cancer, remains a challenge. Here, we perform a parallel deep mutational scan (DMS) of MET intracellular kinase domain in two fusion protein backgrounds: wild type and METΔEx14. Our comparative approach has revealed a critical hydrophobic interaction between a juxtamembrane segment and the kinase αC helix, pointing to differences in regulatory mechanisms between MET and other RTKs. Additionally, we have uncovered a β5 motif that acts as a structural pivot for kinase domain activation in MET and other TAM family of kinases. We also describe a number of previously unknown activating mutations, aiding the effort to annotate driver, passenger, and drug resistance mutations in the MET kinase domain.
format Online
Article
Text
id pubmed-10418267
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Cold Spring Harbor Laboratory
record_format MEDLINE/PubMed
spelling pubmed-104182672023-08-12 Conserved regulatory motifs in the juxtamembrane domain and kinase N-lobe revealed through deep mutational scanning of the MET receptor tyrosine kinase domain Estevam, Gabriella O. Linossi, Edmond M. Macdonald, Christian B. Espinoza, Carla A. Michaud, Jennifer M. Coyote-Maestas, Willow Collisson, Eric A. Jura, Natalia Fraser, James S. bioRxiv Article MET is a receptor tyrosine kinase (RTK) responsible for initiating signaling pathways involved in development and wound repair. MET activation relies on ligand binding to the extracellular receptor, which prompts dimerization, intracellular phosphorylation, and recruitment of associated signaling proteins. Mutations, which are predominantly observed clinically in the intracellular juxtamembrane and kinase domains, can disrupt typical MET regulatory mechanisms. Understanding how juxtamembrane variants, such as exon 14 skipping (METΔEx14), and rare kinase domain mutations can increase signaling, often leading to cancer, remains a challenge. Here, we perform a parallel deep mutational scan (DMS) of MET intracellular kinase domain in two fusion protein backgrounds: wild type and METΔEx14. Our comparative approach has revealed a critical hydrophobic interaction between a juxtamembrane segment and the kinase αC helix, pointing to differences in regulatory mechanisms between MET and other RTKs. Additionally, we have uncovered a β5 motif that acts as a structural pivot for kinase domain activation in MET and other TAM family of kinases. We also describe a number of previously unknown activating mutations, aiding the effort to annotate driver, passenger, and drug resistance mutations in the MET kinase domain. Cold Spring Harbor Laboratory 2023-08-03 /pmc/articles/PMC10418267/ /pubmed/37577651 http://dx.doi.org/10.1101/2023.08.03.551866 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use.
spellingShingle Article
Estevam, Gabriella O.
Linossi, Edmond M.
Macdonald, Christian B.
Espinoza, Carla A.
Michaud, Jennifer M.
Coyote-Maestas, Willow
Collisson, Eric A.
Jura, Natalia
Fraser, James S.
Conserved regulatory motifs in the juxtamembrane domain and kinase N-lobe revealed through deep mutational scanning of the MET receptor tyrosine kinase domain
title Conserved regulatory motifs in the juxtamembrane domain and kinase N-lobe revealed through deep mutational scanning of the MET receptor tyrosine kinase domain
title_full Conserved regulatory motifs in the juxtamembrane domain and kinase N-lobe revealed through deep mutational scanning of the MET receptor tyrosine kinase domain
title_fullStr Conserved regulatory motifs in the juxtamembrane domain and kinase N-lobe revealed through deep mutational scanning of the MET receptor tyrosine kinase domain
title_full_unstemmed Conserved regulatory motifs in the juxtamembrane domain and kinase N-lobe revealed through deep mutational scanning of the MET receptor tyrosine kinase domain
title_short Conserved regulatory motifs in the juxtamembrane domain and kinase N-lobe revealed through deep mutational scanning of the MET receptor tyrosine kinase domain
title_sort conserved regulatory motifs in the juxtamembrane domain and kinase n-lobe revealed through deep mutational scanning of the met receptor tyrosine kinase domain
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10418267/
https://www.ncbi.nlm.nih.gov/pubmed/37577651
http://dx.doi.org/10.1101/2023.08.03.551866
work_keys_str_mv AT estevamgabriellao conservedregulatorymotifsinthejuxtamembranedomainandkinasenloberevealedthroughdeepmutationalscanningofthemetreceptortyrosinekinasedomain
AT linossiedmondm conservedregulatorymotifsinthejuxtamembranedomainandkinasenloberevealedthroughdeepmutationalscanningofthemetreceptortyrosinekinasedomain
AT macdonaldchristianb conservedregulatorymotifsinthejuxtamembranedomainandkinasenloberevealedthroughdeepmutationalscanningofthemetreceptortyrosinekinasedomain
AT espinozacarlaa conservedregulatorymotifsinthejuxtamembranedomainandkinasenloberevealedthroughdeepmutationalscanningofthemetreceptortyrosinekinasedomain
AT michaudjenniferm conservedregulatorymotifsinthejuxtamembranedomainandkinasenloberevealedthroughdeepmutationalscanningofthemetreceptortyrosinekinasedomain
AT coyotemaestaswillow conservedregulatorymotifsinthejuxtamembranedomainandkinasenloberevealedthroughdeepmutationalscanningofthemetreceptortyrosinekinasedomain
AT collissonerica conservedregulatorymotifsinthejuxtamembranedomainandkinasenloberevealedthroughdeepmutationalscanningofthemetreceptortyrosinekinasedomain
AT juranatalia conservedregulatorymotifsinthejuxtamembranedomainandkinasenloberevealedthroughdeepmutationalscanningofthemetreceptortyrosinekinasedomain
AT fraserjamess conservedregulatorymotifsinthejuxtamembranedomainandkinasenloberevealedthroughdeepmutationalscanningofthemetreceptortyrosinekinasedomain