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The focal adhesion-associated proteins DOCK5 and GIT2 comprise a rheostat in control of epithelial invasion

DOCK proteins are guanine nucleotide exchange factors for Rac and Cdc42 GTPases. DOCK1 is the founding member of the family and acts downstream of integrins via the canonical Crk-p130Cas complex to activate Rac GTPases in numerous contexts. In contrast, DOCK5, which possesses the greatest similarity...

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Autores principales: Frank, S R, Köllmann, C P, van Lidth de Jeude, J F, Thiagarajah, J R, Engelholm, L H, Frödin, M, Hansen, S H
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5366285/
https://www.ncbi.nlm.nih.gov/pubmed/27669437
http://dx.doi.org/10.1038/onc.2016.345
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author Frank, S R
Köllmann, C P
van Lidth de Jeude, J F
Thiagarajah, J R
Engelholm, L H
Frödin, M
Hansen, S H
author_facet Frank, S R
Köllmann, C P
van Lidth de Jeude, J F
Thiagarajah, J R
Engelholm, L H
Frödin, M
Hansen, S H
author_sort Frank, S R
collection PubMed
description DOCK proteins are guanine nucleotide exchange factors for Rac and Cdc42 GTPases. DOCK1 is the founding member of the family and acts downstream of integrins via the canonical Crk-p130Cas complex to activate Rac GTPases in numerous contexts. In contrast, DOCK5, which possesses the greatest similarity to DOCK1, remains sparingly studied. Here we establish that DOCK5 has a non-redundant role in regulating motile and invasive capacities of epithelial cells. DOCK1 is constitutively associated with sites of integrin attachment termed focal adhesions (FAs). In contrast, we demonstrate that DOCK5 recruitment to FAs in Hela cells is restricted by GIT2, an established regulator of FA signaling. We determine that GIT2 is targeted to FAs in response to Rho-ROCK signaling and actomyosin contractility. Accordingly, inhibition of ROCK activity or MLC function promotes enrichment of DOCK5 in membrane protrusions and nascent cell–substratum adhesions. We further demonstrate that GIT2 inhibits the interaction of DOCK5 with Crk. Moreover, we show that depletion of GIT2 promotes DOCK5-dependent activation of the Crk-p130Cas signaling cascade to promote Rac1-mediated lamellipodial protrusion and FA turnover. The antagonism between GIT2 and DOCK5 extends to non-transformed MCF10A mammary epithelial cells, with DOCK5 ‘dialing-up' and GIT2 ‘dialing-down' invasiveness. Finally, we determine that DOCK5 inhibition attenuates invasion and metastasis of MDA-MB-231 cells and prolongs life span of mice injected with these cells. Collectively, our work identifies DOCK5 as a key regulator of epithelial invasion and metastasis, and demonstrates that suppression of DOCK5 by GIT2 represents a previously unappreciated mechanism for coordination of Rho and Rac GTPases.
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spelling pubmed-53662852017-03-27 The focal adhesion-associated proteins DOCK5 and GIT2 comprise a rheostat in control of epithelial invasion Frank, S R Köllmann, C P van Lidth de Jeude, J F Thiagarajah, J R Engelholm, L H Frödin, M Hansen, S H Oncogene Original Article DOCK proteins are guanine nucleotide exchange factors for Rac and Cdc42 GTPases. DOCK1 is the founding member of the family and acts downstream of integrins via the canonical Crk-p130Cas complex to activate Rac GTPases in numerous contexts. In contrast, DOCK5, which possesses the greatest similarity to DOCK1, remains sparingly studied. Here we establish that DOCK5 has a non-redundant role in regulating motile and invasive capacities of epithelial cells. DOCK1 is constitutively associated with sites of integrin attachment termed focal adhesions (FAs). In contrast, we demonstrate that DOCK5 recruitment to FAs in Hela cells is restricted by GIT2, an established regulator of FA signaling. We determine that GIT2 is targeted to FAs in response to Rho-ROCK signaling and actomyosin contractility. Accordingly, inhibition of ROCK activity or MLC function promotes enrichment of DOCK5 in membrane protrusions and nascent cell–substratum adhesions. We further demonstrate that GIT2 inhibits the interaction of DOCK5 with Crk. Moreover, we show that depletion of GIT2 promotes DOCK5-dependent activation of the Crk-p130Cas signaling cascade to promote Rac1-mediated lamellipodial protrusion and FA turnover. The antagonism between GIT2 and DOCK5 extends to non-transformed MCF10A mammary epithelial cells, with DOCK5 ‘dialing-up' and GIT2 ‘dialing-down' invasiveness. Finally, we determine that DOCK5 inhibition attenuates invasion and metastasis of MDA-MB-231 cells and prolongs life span of mice injected with these cells. Collectively, our work identifies DOCK5 as a key regulator of epithelial invasion and metastasis, and demonstrates that suppression of DOCK5 by GIT2 represents a previously unappreciated mechanism for coordination of Rho and Rac GTPases. Nature Publishing Group 2017-03-30 2016-09-26 /pmc/articles/PMC5366285/ /pubmed/27669437 http://dx.doi.org/10.1038/onc.2016.345 Text en Copyright © 2017 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/
spellingShingle Original Article
Frank, S R
Köllmann, C P
van Lidth de Jeude, J F
Thiagarajah, J R
Engelholm, L H
Frödin, M
Hansen, S H
The focal adhesion-associated proteins DOCK5 and GIT2 comprise a rheostat in control of epithelial invasion
title The focal adhesion-associated proteins DOCK5 and GIT2 comprise a rheostat in control of epithelial invasion
title_full The focal adhesion-associated proteins DOCK5 and GIT2 comprise a rheostat in control of epithelial invasion
title_fullStr The focal adhesion-associated proteins DOCK5 and GIT2 comprise a rheostat in control of epithelial invasion
title_full_unstemmed The focal adhesion-associated proteins DOCK5 and GIT2 comprise a rheostat in control of epithelial invasion
title_short The focal adhesion-associated proteins DOCK5 and GIT2 comprise a rheostat in control of epithelial invasion
title_sort focal adhesion-associated proteins dock5 and git2 comprise a rheostat in control of epithelial invasion
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5366285/
https://www.ncbi.nlm.nih.gov/pubmed/27669437
http://dx.doi.org/10.1038/onc.2016.345
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