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Non-canonical dynamic mechanisms of interaction between the p66Shc protein and Met receptor

Met receptor tyrosine kinase (RTK) is known to bind to the three distinct protein isoforms encoded by the ShcA (Shc) gene. Structure–function studies have unveiled critical roles for p52Shc-dependent signalling pathways in Met-regulated biological functions. The molecular basis of the interaction be...

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Autores principales: Landry, Mélissa, Pomerleau, Véronique, Saucier, Caroline
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4888465/
https://www.ncbi.nlm.nih.gov/pubmed/27048591
http://dx.doi.org/10.1042/BCJ20160249
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author Landry, Mélissa
Pomerleau, Véronique
Saucier, Caroline
author_facet Landry, Mélissa
Pomerleau, Véronique
Saucier, Caroline
author_sort Landry, Mélissa
collection PubMed
description Met receptor tyrosine kinase (RTK) is known to bind to the three distinct protein isoforms encoded by the ShcA (Shc) gene. Structure–function studies have unveiled critical roles for p52Shc-dependent signalling pathways in Met-regulated biological functions. The molecular basis of the interaction between the Met and p52Shc proteins is well-defined, but not for the longest protein isoform, p66Shc. In the present study, co-immunoprecipitation assays were performed in human embryonic kidney 293 (HEK293) cells, transiently co-transfected with Met and p66Shc mutants, in order to define the molecular determinants involved in mediating Met–p66Shc interaction. Our results show that p66Shc interacts constitutively with the receptor Met, and the Grb2 (growth factor receptor-bound protein-2) and Gab1 (Grb2-associated binder-1) adaptor proteins. Although its phosphotyrosine-binding domain (PTB) and Src homology 2 (SH2) domains co-ordinate p66Shc binding to non-activated Met receptor, these phosphotyrosine-binding modules, and its collagen homology domain 2 (CH2) region, exert negative constraints. In contrast, p66Shc interaction with the activated Met depends mainly on the integrity of its PTB domain, and to a lesser extent of its SH2 domain. Even though not required for the recruitment of p66Shc, tyrosine phosphorylation of p66Shc by activated Met enhances these interactions by mechanisms not reliant on the integrity of the Met multisubstrate-binding site. In turn, this increases phosphotyrosine-dependent p66Shc–Grb2–Gab1 complex formation away from the receptor, while blocking Grb2 and Gab1 recruitment to activated Met. In conclusion, we identify, for the first time, a novel non-canonical dynamic mode of interaction between Met and the p66 protein isoform of Shc and its effects on rewiring binding effector complexes according to the activation state of the receptor.
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spelling pubmed-48884652016-06-08 Non-canonical dynamic mechanisms of interaction between the p66Shc protein and Met receptor Landry, Mélissa Pomerleau, Véronique Saucier, Caroline Biochem J Research Articles Met receptor tyrosine kinase (RTK) is known to bind to the three distinct protein isoforms encoded by the ShcA (Shc) gene. Structure–function studies have unveiled critical roles for p52Shc-dependent signalling pathways in Met-regulated biological functions. The molecular basis of the interaction between the Met and p52Shc proteins is well-defined, but not for the longest protein isoform, p66Shc. In the present study, co-immunoprecipitation assays were performed in human embryonic kidney 293 (HEK293) cells, transiently co-transfected with Met and p66Shc mutants, in order to define the molecular determinants involved in mediating Met–p66Shc interaction. Our results show that p66Shc interacts constitutively with the receptor Met, and the Grb2 (growth factor receptor-bound protein-2) and Gab1 (Grb2-associated binder-1) adaptor proteins. Although its phosphotyrosine-binding domain (PTB) and Src homology 2 (SH2) domains co-ordinate p66Shc binding to non-activated Met receptor, these phosphotyrosine-binding modules, and its collagen homology domain 2 (CH2) region, exert negative constraints. In contrast, p66Shc interaction with the activated Met depends mainly on the integrity of its PTB domain, and to a lesser extent of its SH2 domain. Even though not required for the recruitment of p66Shc, tyrosine phosphorylation of p66Shc by activated Met enhances these interactions by mechanisms not reliant on the integrity of the Met multisubstrate-binding site. In turn, this increases phosphotyrosine-dependent p66Shc–Grb2–Gab1 complex formation away from the receptor, while blocking Grb2 and Gab1 recruitment to activated Met. In conclusion, we identify, for the first time, a novel non-canonical dynamic mode of interaction between Met and the p66 protein isoform of Shc and its effects on rewiring binding effector complexes according to the activation state of the receptor. Portland Press Ltd. 2016-05-27 2016-06-01 /pmc/articles/PMC4888465/ /pubmed/27048591 http://dx.doi.org/10.1042/BCJ20160249 Text en © 2016 The Author(s). published by Portland Press Limited on behalf of the Biochemical Society
spellingShingle Research Articles
Landry, Mélissa
Pomerleau, Véronique
Saucier, Caroline
Non-canonical dynamic mechanisms of interaction between the p66Shc protein and Met receptor
title Non-canonical dynamic mechanisms of interaction between the p66Shc protein and Met receptor
title_full Non-canonical dynamic mechanisms of interaction between the p66Shc protein and Met receptor
title_fullStr Non-canonical dynamic mechanisms of interaction between the p66Shc protein and Met receptor
title_full_unstemmed Non-canonical dynamic mechanisms of interaction between the p66Shc protein and Met receptor
title_short Non-canonical dynamic mechanisms of interaction between the p66Shc protein and Met receptor
title_sort non-canonical dynamic mechanisms of interaction between the p66shc protein and met receptor
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4888465/
https://www.ncbi.nlm.nih.gov/pubmed/27048591
http://dx.doi.org/10.1042/BCJ20160249
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