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Phosphorylation of Serine 106 in Asef2 Regulates Cell Migration and Adhesion Turnover

[Image: see text] Asef2, a 652-amino acid protein, is a guanine nucleotide exchange factor (GEF) that regulates cell migration and other processes via activation of Rho family GTPases, including Rac. Binding of the tumor suppressor adenomatous polyposis coli (APC) to Asef2 is known to induce its GEF...

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Autores principales: Evans, J. Corey, Hines, Kelly M., Forsythe, Jay G., Erdogan, Begum, Shi, Mingjian, Hill, Salisha, Rose, Kristie L., McLean, John A., Webb, Donna J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2014
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4084842/
https://www.ncbi.nlm.nih.gov/pubmed/24874604
http://dx.doi.org/10.1021/pr5001384
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author Evans, J. Corey
Hines, Kelly M.
Forsythe, Jay G.
Erdogan, Begum
Shi, Mingjian
Hill, Salisha
Rose, Kristie L.
McLean, John A.
Webb, Donna J.
author_facet Evans, J. Corey
Hines, Kelly M.
Forsythe, Jay G.
Erdogan, Begum
Shi, Mingjian
Hill, Salisha
Rose, Kristie L.
McLean, John A.
Webb, Donna J.
author_sort Evans, J. Corey
collection PubMed
description [Image: see text] Asef2, a 652-amino acid protein, is a guanine nucleotide exchange factor (GEF) that regulates cell migration and other processes via activation of Rho family GTPases, including Rac. Binding of the tumor suppressor adenomatous polyposis coli (APC) to Asef2 is known to induce its GEF activity; however, little is currently known about other modes of Asef2 regulation. Here, we investigated the role of phosphorylation in regulating Asef2 activity and function. Using high-resolution mass spectrometry (MS) and tandem mass spectrometry (MS/MS), we obtained complete coverage of all phosphorylatable residues and identified six phosphorylation sites. One of these, serine 106 (S106), was particularly intriguing as a potential regulator of Asef2 activity because of its location within the APC-binding domain. Interestingly, mutation of this serine to alanine (S106A), a non-phosphorylatable analogue, greatly diminished the ability of Asef2 to activate Rac, while a phosphomimetic mutation (serine to aspartic acid, S106D) enhanced Rac activation. Furthermore, expression of these mutants in HT1080 cells demonstrated that phosphorylation of S106 is critical for Asef2-promoted migration and for cell-matrix adhesion assembly and disassembly (adhesion turnover), which is a process that facilitates efficient migration. Collectively, our results show that phosphorylation of S106 modulates Asef2 GEF activity and Asef2-mediated cell migration and adhesion turnover.
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spelling pubmed-40848422015-05-29 Phosphorylation of Serine 106 in Asef2 Regulates Cell Migration and Adhesion Turnover Evans, J. Corey Hines, Kelly M. Forsythe, Jay G. Erdogan, Begum Shi, Mingjian Hill, Salisha Rose, Kristie L. McLean, John A. Webb, Donna J. J Proteome Res [Image: see text] Asef2, a 652-amino acid protein, is a guanine nucleotide exchange factor (GEF) that regulates cell migration and other processes via activation of Rho family GTPases, including Rac. Binding of the tumor suppressor adenomatous polyposis coli (APC) to Asef2 is known to induce its GEF activity; however, little is currently known about other modes of Asef2 regulation. Here, we investigated the role of phosphorylation in regulating Asef2 activity and function. Using high-resolution mass spectrometry (MS) and tandem mass spectrometry (MS/MS), we obtained complete coverage of all phosphorylatable residues and identified six phosphorylation sites. One of these, serine 106 (S106), was particularly intriguing as a potential regulator of Asef2 activity because of its location within the APC-binding domain. Interestingly, mutation of this serine to alanine (S106A), a non-phosphorylatable analogue, greatly diminished the ability of Asef2 to activate Rac, while a phosphomimetic mutation (serine to aspartic acid, S106D) enhanced Rac activation. Furthermore, expression of these mutants in HT1080 cells demonstrated that phosphorylation of S106 is critical for Asef2-promoted migration and for cell-matrix adhesion assembly and disassembly (adhesion turnover), which is a process that facilitates efficient migration. Collectively, our results show that phosphorylation of S106 modulates Asef2 GEF activity and Asef2-mediated cell migration and adhesion turnover. American Chemical Society 2014-05-29 2014-07-03 /pmc/articles/PMC4084842/ /pubmed/24874604 http://dx.doi.org/10.1021/pr5001384 Text en Copyright © 2014 American Chemical Society Terms of Use (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html)
spellingShingle Evans, J. Corey
Hines, Kelly M.
Forsythe, Jay G.
Erdogan, Begum
Shi, Mingjian
Hill, Salisha
Rose, Kristie L.
McLean, John A.
Webb, Donna J.
Phosphorylation of Serine 106 in Asef2 Regulates Cell Migration and Adhesion Turnover
title Phosphorylation of Serine 106 in Asef2 Regulates Cell Migration and Adhesion Turnover
title_full Phosphorylation of Serine 106 in Asef2 Regulates Cell Migration and Adhesion Turnover
title_fullStr Phosphorylation of Serine 106 in Asef2 Regulates Cell Migration and Adhesion Turnover
title_full_unstemmed Phosphorylation of Serine 106 in Asef2 Regulates Cell Migration and Adhesion Turnover
title_short Phosphorylation of Serine 106 in Asef2 Regulates Cell Migration and Adhesion Turnover
title_sort phosphorylation of serine 106 in asef2 regulates cell migration and adhesion turnover
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4084842/
https://www.ncbi.nlm.nih.gov/pubmed/24874604
http://dx.doi.org/10.1021/pr5001384
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