PP2A phosphatase suppresses function of the mesenchymal invasion regulator NEDD9

Bradbury, P; Mahmassani, M; Zhong, J; Turner, K; Paul, A; Verrills, NM; O'Neill, GM

PP2A phosphatase suppresses function of the mesenchymal invasion regulator NEDD9

Bradbury, P

Mahmassani, M Zhong, J Turner, K Paul, A Verrills, NM O'Neill, GM

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Publication Type:: Journal Article
Citation:: Biochimica et Biophysica Acta - Molecular Cell Research, 2012, 1823 (2), pp. 290 - 297
Issue Date:: 2012-02-01

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Field	Value	Language
dc.contributor.author	Bradbury, P https://orcid.org/0000-0001-6360-9591	en_US
dc.contributor.author	Mahmassani, M	en_US
dc.contributor.author	Zhong, J	en_US
dc.contributor.author	Turner, K	en_US
dc.contributor.author	Paul, A	en_US
dc.contributor.author	Verrills, NM	en_US
dc.contributor.author	O'Neill, GM	en_US
dc.date.available	2011-10-18	en_US
dc.date.issued	2012-02-01	en_US
dc.identifier.citation	Biochimica et Biophysica Acta - Molecular Cell Research, 2012, 1823 (2), pp. 290 - 297	en_US
dc.identifier.issn	0167-4889	en_US
dc.identifier.uri	http://hdl.handle.net/10453/115494
dc.description.abstract	The mesenchymal mode of cancer cell invasion characterized by active adhesion turnover and a polarized actin cytoskeleton, is critically regulated by the adaptor protein NEDD9/HEF1/Cas-L. While it is known that NEDD9 is subject to extensive phosphorylation modification, the molecules that determine NEDD9 phosphorylation to stimulate adhesion turnover and mesenchymal cell morphologies are currently unknown. Earlier studies have suggested that the serine/threonine phosphatase PP2A regulates interconversion between a low molecular mass NEDD9 phosphoform and higher molecular mass phosphoforms. However, previous studies have used chemical inhibitors to block PP2A activity. In the present study we therefore aimed to specifically inhibit PP2A activity via siRNA and dominant negative approaches to investigate the effect of PP2A on interconversion between 115. kDa and 105. kDa NEDD9 and determine the functional consequence of PP2A activity for NEDD9 function. Strikingly, we find that while the phosphatase inhibitor Calyculin A indeed abrogates detachment-induced dephosphorylation of the 115. kDa NEDD9 phosphoform, PP2A depletion does not inhibit 115. kDa to 105. kDa interconversion. Our data suggest instead that PP2A targets discrete NEDD9 phosphorylation modifications separate to the events that mediate interconversion between the two forms. Functionally, PP2A depletion increases NEDD9 mediated cell spreading and mutation of S369 in the serine-rich region of NEDD9 to aspartate mimics this effect. Importantly, mutation of S369 to alanine abrogates the ability of dominant negative PP2A to increase NEDD9-mediated cell spreading. Collectively, our data reveal that the tumour suppressor PP2A may act via S369 to regulated NEDD9-mediated cell spreading. © 2011.	en_US
dc.relation.ispartof	Biochimica et Biophysica Acta - Molecular Cell Research	en_US
dc.relation.isbasedon	10.1016/j.bbamcr.2011.10.011	en_US
dc.subject.mesh	Cell Line	en_US
dc.subject.mesh	Mesoderm	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Oxazoles	en_US
dc.subject.mesh	Adaptor Proteins, Signal Transducing	en_US
dc.subject.mesh	Leupeptins	en_US
dc.subject.mesh	Phosphoproteins	en_US
dc.subject.mesh	Protein Isoforms	en_US
dc.subject.mesh	Recombinant Fusion Proteins	en_US
dc.subject.mesh	Cysteine Proteinase Inhibitors	en_US
dc.subject.mesh	Cell Movement	en_US
dc.subject.mesh	RNA Interference	en_US
dc.subject.mesh	Protein Phosphatase 2	en_US
dc.title	PP2A phosphatase suppresses function of the mesenchymal invasion regulator NEDD9	en_US
dc.type	Journal Article
utslib.citation.volume	2	en_US
utslib.citation.volume	1823	en_US
utslib.for	02 Physical Sciences	en_US
utslib.for	06 Biological Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology/School of Biomedical Engineering
utslib.copyright.status	closed_access
pubs.issue	2	en_US
pubs.publication-status	Published	en_US
pubs.volume	1823	en_US

Abstract:

The mesenchymal mode of cancer cell invasion characterized by active adhesion turnover and a polarized actin cytoskeleton, is critically regulated by the adaptor protein NEDD9/HEF1/Cas-L. While it is known that NEDD9 is subject to extensive phosphorylation modification, the molecules that determine NEDD9 phosphorylation to stimulate adhesion turnover and mesenchymal cell morphologies are currently unknown. Earlier studies have suggested that the serine/threonine phosphatase PP2A regulates interconversion between a low molecular mass NEDD9 phosphoform and higher molecular mass phosphoforms. However, previous studies have used chemical inhibitors to block PP2A activity. In the present study we therefore aimed to specifically inhibit PP2A activity via siRNA and dominant negative approaches to investigate the effect of PP2A on interconversion between 115. kDa and 105. kDa NEDD9 and determine the functional consequence of PP2A activity for NEDD9 function. Strikingly, we find that while the phosphatase inhibitor Calyculin A indeed abrogates detachment-induced dephosphorylation of the 115. kDa NEDD9 phosphoform, PP2A depletion does not inhibit 115. kDa to 105. kDa interconversion. Our data suggest instead that PP2A targets discrete NEDD9 phosphorylation modifications separate to the events that mediate interconversion between the two forms. Functionally, PP2A depletion increases NEDD9 mediated cell spreading and mutation of S369 in the serine-rich region of NEDD9 to aspartate mimics this effect. Importantly, mutation of S369 to alanine abrogates the ability of dominant negative PP2A to increase NEDD9-mediated cell spreading. Collectively, our data reveal that the tumour suppressor PP2A may act via S369 to regulated NEDD9-mediated cell spreading. © 2011.

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http://hdl.handle.net/10453/115494