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A Screen for PKN3 Substrates Reveals an Activating Phosphorylation of ARHGAP18

Protein kinase N3 (PKN3) is a serine/threonine kinase implicated in tumor progression of multiple cancer types, however, its substrates and effector proteins still remain largely understudied. In the present work we aimed to identify novel PKN3 substrates in a phosphoproteomic screen using analog se...

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Detalles Bibliográficos
Autores principales: Dibus, Michal, Brábek, Jan, Rösel, Daniel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7594087/
https://www.ncbi.nlm.nih.gov/pubmed/33092266
http://dx.doi.org/10.3390/ijms21207769
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author Dibus, Michal
Brábek, Jan
Rösel, Daniel
author_facet Dibus, Michal
Brábek, Jan
Rösel, Daniel
author_sort Dibus, Michal
collection PubMed
description Protein kinase N3 (PKN3) is a serine/threonine kinase implicated in tumor progression of multiple cancer types, however, its substrates and effector proteins still remain largely understudied. In the present work we aimed to identify novel PKN3 substrates in a phosphoproteomic screen using analog sensitive PKN3. Among the identified putative substrates we selected ARHGAP18, a protein from RhoGAP family, for validation of the screen and further study. We confirmed that PKN3 can phosphorylate ARHGAP18 in vitro and we also characterized the interaction of the two proteins, which is mediated via the N-terminal part of ARHGAP18. We present strong evidence that PKN3-ARHGAP18 interaction is increased upon ARHGAP18 phosphorylation and that the phosphorylation of ARHGAP18 by PKN3 enhances its GAP domain activity and contributes to negative regulation of active RhoA. Taken together, we identified new set of potential PKN3 substrates and revealed a new negative feedback regulatory mechanism of Rho signaling mediated by PKN3-induced ARHGAP18 activation.
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spelling pubmed-75940872020-10-30 A Screen for PKN3 Substrates Reveals an Activating Phosphorylation of ARHGAP18 Dibus, Michal Brábek, Jan Rösel, Daniel Int J Mol Sci Article Protein kinase N3 (PKN3) is a serine/threonine kinase implicated in tumor progression of multiple cancer types, however, its substrates and effector proteins still remain largely understudied. In the present work we aimed to identify novel PKN3 substrates in a phosphoproteomic screen using analog sensitive PKN3. Among the identified putative substrates we selected ARHGAP18, a protein from RhoGAP family, for validation of the screen and further study. We confirmed that PKN3 can phosphorylate ARHGAP18 in vitro and we also characterized the interaction of the two proteins, which is mediated via the N-terminal part of ARHGAP18. We present strong evidence that PKN3-ARHGAP18 interaction is increased upon ARHGAP18 phosphorylation and that the phosphorylation of ARHGAP18 by PKN3 enhances its GAP domain activity and contributes to negative regulation of active RhoA. Taken together, we identified new set of potential PKN3 substrates and revealed a new negative feedback regulatory mechanism of Rho signaling mediated by PKN3-induced ARHGAP18 activation. MDPI 2020-10-20 /pmc/articles/PMC7594087/ /pubmed/33092266 http://dx.doi.org/10.3390/ijms21207769 Text en © 2020 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
Dibus, Michal
Brábek, Jan
Rösel, Daniel
A Screen for PKN3 Substrates Reveals an Activating Phosphorylation of ARHGAP18
title A Screen for PKN3 Substrates Reveals an Activating Phosphorylation of ARHGAP18
title_full A Screen for PKN3 Substrates Reveals an Activating Phosphorylation of ARHGAP18
title_fullStr A Screen for PKN3 Substrates Reveals an Activating Phosphorylation of ARHGAP18
title_full_unstemmed A Screen for PKN3 Substrates Reveals an Activating Phosphorylation of ARHGAP18
title_short A Screen for PKN3 Substrates Reveals an Activating Phosphorylation of ARHGAP18
title_sort screen for pkn3 substrates reveals an activating phosphorylation of arhgap18
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7594087/
https://www.ncbi.nlm.nih.gov/pubmed/33092266
http://dx.doi.org/10.3390/ijms21207769
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