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Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2

Discoidin, CUB, and LCCL Domain-containing (DCBLD) 2 is a neuropilin-like transmembrane scaffolding receptor with known and anticipated roles in vascular remodeling and neuronal positioning. DCBLD2 is also upregulated in several cancers and can drive glioblastomas downstream of activated Epidermal G...

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Autores principales: Schmoker, Anna M., Weinert, Jaye L., Kellett, Kyle J., Johnson, Hannah E., Joy, Ryan M., Weir, Marion E., Ebert, Alicia M., Ballif, Bryan A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6029619/
https://www.ncbi.nlm.nih.gov/pubmed/29025973
http://dx.doi.org/10.1042/BCJ20170615
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author Schmoker, Anna M.
Weinert, Jaye L.
Kellett, Kyle J.
Johnson, Hannah E.
Joy, Ryan M.
Weir, Marion E.
Ebert, Alicia M.
Ballif, Bryan A.
author_facet Schmoker, Anna M.
Weinert, Jaye L.
Kellett, Kyle J.
Johnson, Hannah E.
Joy, Ryan M.
Weir, Marion E.
Ebert, Alicia M.
Ballif, Bryan A.
author_sort Schmoker, Anna M.
collection PubMed
description Discoidin, CUB, and LCCL Domain-containing (DCBLD) 2 is a neuropilin-like transmembrane scaffolding receptor with known and anticipated roles in vascular remodeling and neuronal positioning. DCBLD2 is also upregulated in several cancers and can drive glioblastomas downstream of activated Epidermal Growth Factor Receptor. While a few studies have shown either a positive or negative role for DCBLD2 in regulating growth factor receptor signaling, little is known about the conserved signaling features of DCBLD family members that drive their molecular activities. We previously identified DCBLD2 tyrosine phosphorylation sites in intracellular YxxP motifs that are required for the phosphorylation-dependent binding of the signaling adaptors CRK and CRKL (CT10 regulator of kinase and CRK-Like). These intracellular YxxP motifs are highly conserved across vertebrates and between DCBLD family members. Here, we demonstrate that, as for DCBLD2, DCBLD1 YxxP motifs are required for CRKL-SH2 binding. We report Src family kinases (SFKs) and Abl differentially promote the interaction between the CRKL-SH2 domain and DCBLD1 and DCBLD2, and while SFKs and Abl each promotes DCBLD1 and DCBLD2 binding to the CRKL-SH2 domain, the effect of Abl is more pronounced for DCBLD1. Using high performance liquid chromatography coupled with tandem mass spectrometry, we quantified phosphorylation at several YxxP sites in DCBLD1 and DCBLD2, mapping site-specific preferences for SFKs and Abl. Together these data provide a platform to decipher the signaling mechanisms by which these novel receptors drive their biological activities.
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spelling pubmed-60296192018-11-21 Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2 Schmoker, Anna M. Weinert, Jaye L. Kellett, Kyle J. Johnson, Hannah E. Joy, Ryan M. Weir, Marion E. Ebert, Alicia M. Ballif, Bryan A. Biochem J Article Discoidin, CUB, and LCCL Domain-containing (DCBLD) 2 is a neuropilin-like transmembrane scaffolding receptor with known and anticipated roles in vascular remodeling and neuronal positioning. DCBLD2 is also upregulated in several cancers and can drive glioblastomas downstream of activated Epidermal Growth Factor Receptor. While a few studies have shown either a positive or negative role for DCBLD2 in regulating growth factor receptor signaling, little is known about the conserved signaling features of DCBLD family members that drive their molecular activities. We previously identified DCBLD2 tyrosine phosphorylation sites in intracellular YxxP motifs that are required for the phosphorylation-dependent binding of the signaling adaptors CRK and CRKL (CT10 regulator of kinase and CRK-Like). These intracellular YxxP motifs are highly conserved across vertebrates and between DCBLD family members. Here, we demonstrate that, as for DCBLD2, DCBLD1 YxxP motifs are required for CRKL-SH2 binding. We report Src family kinases (SFKs) and Abl differentially promote the interaction between the CRKL-SH2 domain and DCBLD1 and DCBLD2, and while SFKs and Abl each promotes DCBLD1 and DCBLD2 binding to the CRKL-SH2 domain, the effect of Abl is more pronounced for DCBLD1. Using high performance liquid chromatography coupled with tandem mass spectrometry, we quantified phosphorylation at several YxxP sites in DCBLD1 and DCBLD2, mapping site-specific preferences for SFKs and Abl. Together these data provide a platform to decipher the signaling mechanisms by which these novel receptors drive their biological activities. 2017-11-21 /pmc/articles/PMC6029619/ /pubmed/29025973 http://dx.doi.org/10.1042/BCJ20170615 Text en http://creativecommons.org/licenses/by/4.0/ Use of open access articles is permitted based on the terms of the specific Creative Commons Licence under which the article is published. Archiving of non-open access articles is permitted in accordance with the Archiving Policy of Portland Press (http://www.portlandpresspublishing.com/content/open-access-policy#Archiving).
spellingShingle Article
Schmoker, Anna M.
Weinert, Jaye L.
Kellett, Kyle J.
Johnson, Hannah E.
Joy, Ryan M.
Weir, Marion E.
Ebert, Alicia M.
Ballif, Bryan A.
Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2
title Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2
title_full Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2
title_fullStr Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2
title_full_unstemmed Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2
title_short Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2
title_sort dynamic multi-site phosphorylation by fyn and abl drives the interaction between crkl and the novel scaffolding receptors dcbld1 and dcbld2
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6029619/
https://www.ncbi.nlm.nih.gov/pubmed/29025973
http://dx.doi.org/10.1042/BCJ20170615
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