Cargando…

Fyn Regulates Binding Partners of Cyclic-AMP Dependent Protein Kinase A

The cAMP-dependent protein kinase A (PKA) is a serine/threonine kinase involved in many fundamental cellular processes, including migration and proliferation. Recently, we found that the Src family kinase Fyn phosphorylates the catalytic subunit of PKA (PKA-C) at Y69, thereby increasing PKA kinase a...

Descripción completa

Detalles Bibliográficos
Autores principales: Schmoker, Anna M., Barritt, Samuel A., Weir, Marion E., Mann, Jacqueline E., Hogan, Tyler C., Ballif, Bryan A., Deming, Paula B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6313912/
https://www.ncbi.nlm.nih.gov/pubmed/30274258
http://dx.doi.org/10.3390/proteomes6040037
_version_ 1783384041402138624
author Schmoker, Anna M.
Barritt, Samuel A.
Weir, Marion E.
Mann, Jacqueline E.
Hogan, Tyler C.
Ballif, Bryan A.
Deming, Paula B.
author_facet Schmoker, Anna M.
Barritt, Samuel A.
Weir, Marion E.
Mann, Jacqueline E.
Hogan, Tyler C.
Ballif, Bryan A.
Deming, Paula B.
author_sort Schmoker, Anna M.
collection PubMed
description The cAMP-dependent protein kinase A (PKA) is a serine/threonine kinase involved in many fundamental cellular processes, including migration and proliferation. Recently, we found that the Src family kinase Fyn phosphorylates the catalytic subunit of PKA (PKA-C) at Y69, thereby increasing PKA kinase activity. We also showed that Fyn induced the phosphorylation of cellular proteins within the PKA preferred target motif. This led to the hypothesis that Fyn could affect proteins in complex with PKA. To test this, we employed a quantitative mass spectrometry approach to identify Fyn-dependent binding partners in complex with PKA-C. We found Fyn enhanced the binding of PKA-C to several cytoskeletal regulators that localize to the centrosome and Golgi apparatus. Three of these Fyn-induced PKA interactors, AKAP9, PDE4DIP, and CDK5RAP2, were validated biochemically and were shown to exist in complex with Fyn and PKA in a glioblastoma cell line. Intriguingly, the complexes formed between PKA-C and these known AKAPs were dependent upon Fyn catalytic activity and expression levels. In addition, we identified Fyn-regulated phosphorylation sites on proteins in complex with PKA-C. We also identified and biochemically validated a novel PKA-C interactor, LARP4, which complexed with PKA in the absence of Fyn. These results demonstrate the ability of Fyn to influence the docking of PKA to specific cellular scaffolds and suggest that Fyn may affect the downstream substrates targeted by PKA.
format Online
Article
Text
id pubmed-6313912
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-63139122019-01-07 Fyn Regulates Binding Partners of Cyclic-AMP Dependent Protein Kinase A Schmoker, Anna M. Barritt, Samuel A. Weir, Marion E. Mann, Jacqueline E. Hogan, Tyler C. Ballif, Bryan A. Deming, Paula B. Proteomes Article The cAMP-dependent protein kinase A (PKA) is a serine/threonine kinase involved in many fundamental cellular processes, including migration and proliferation. Recently, we found that the Src family kinase Fyn phosphorylates the catalytic subunit of PKA (PKA-C) at Y69, thereby increasing PKA kinase activity. We also showed that Fyn induced the phosphorylation of cellular proteins within the PKA preferred target motif. This led to the hypothesis that Fyn could affect proteins in complex with PKA. To test this, we employed a quantitative mass spectrometry approach to identify Fyn-dependent binding partners in complex with PKA-C. We found Fyn enhanced the binding of PKA-C to several cytoskeletal regulators that localize to the centrosome and Golgi apparatus. Three of these Fyn-induced PKA interactors, AKAP9, PDE4DIP, and CDK5RAP2, were validated biochemically and were shown to exist in complex with Fyn and PKA in a glioblastoma cell line. Intriguingly, the complexes formed between PKA-C and these known AKAPs were dependent upon Fyn catalytic activity and expression levels. In addition, we identified Fyn-regulated phosphorylation sites on proteins in complex with PKA-C. We also identified and biochemically validated a novel PKA-C interactor, LARP4, which complexed with PKA in the absence of Fyn. These results demonstrate the ability of Fyn to influence the docking of PKA to specific cellular scaffolds and suggest that Fyn may affect the downstream substrates targeted by PKA. MDPI 2018-09-29 /pmc/articles/PMC6313912/ /pubmed/30274258 http://dx.doi.org/10.3390/proteomes6040037 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Schmoker, Anna M.
Barritt, Samuel A.
Weir, Marion E.
Mann, Jacqueline E.
Hogan, Tyler C.
Ballif, Bryan A.
Deming, Paula B.
Fyn Regulates Binding Partners of Cyclic-AMP Dependent Protein Kinase A
title Fyn Regulates Binding Partners of Cyclic-AMP Dependent Protein Kinase A
title_full Fyn Regulates Binding Partners of Cyclic-AMP Dependent Protein Kinase A
title_fullStr Fyn Regulates Binding Partners of Cyclic-AMP Dependent Protein Kinase A
title_full_unstemmed Fyn Regulates Binding Partners of Cyclic-AMP Dependent Protein Kinase A
title_short Fyn Regulates Binding Partners of Cyclic-AMP Dependent Protein Kinase A
title_sort fyn regulates binding partners of cyclic-amp dependent protein kinase a
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6313912/
https://www.ncbi.nlm.nih.gov/pubmed/30274258
http://dx.doi.org/10.3390/proteomes6040037
work_keys_str_mv AT schmokerannam fynregulatesbindingpartnersofcyclicampdependentproteinkinasea
AT barrittsamuela fynregulatesbindingpartnersofcyclicampdependentproteinkinasea
AT weirmarione fynregulatesbindingpartnersofcyclicampdependentproteinkinasea
AT mannjacquelinee fynregulatesbindingpartnersofcyclicampdependentproteinkinasea
AT hogantylerc fynregulatesbindingpartnersofcyclicampdependentproteinkinasea
AT ballifbryana fynregulatesbindingpartnersofcyclicampdependentproteinkinasea
AT demingpaulab fynregulatesbindingpartnersofcyclicampdependentproteinkinasea