The focal adhesion targeting domain of p130Cas confers a mechanosensing function

Bradbury, PM; Turner, K; Mitchell, C; Griffin, KR; Middlemiss, S; Lau, L; Dagg, R; Taran, E; Cooper-White, J; Fabry, B; O'Neill, GM

The focal adhesion targeting domain of p130Cas confers a mechanosensing function

Bradbury, PM

Turner, K Mitchell, C Griffin, KR Middlemiss, S Lau, L Dagg, R Taran, E Cooper-White, J Fabry, B O'Neill, GM

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Publication Type:: Journal Article
Citation:: Journal of Cell Science, 2017, 130 (7), pp. 1263 - 1273
Issue Date:: 2017-04-01

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Field	Value	Language
dc.contributor.author	Bradbury, PM https://orcid.org/0000-0001-6360-9591	en_US
dc.contributor.author	Turner, K	en_US
dc.contributor.author	Mitchell, C	en_US
dc.contributor.author	Griffin, KR	en_US
dc.contributor.author	Middlemiss, S	en_US
dc.contributor.author	Lau, L	en_US
dc.contributor.author	Dagg, R	en_US
dc.contributor.author	Taran, E	en_US
dc.contributor.author	Cooper-White, J	en_US
dc.contributor.author	Fabry, B	en_US
dc.contributor.author	O'Neill, GM	en_US
dc.date.available	2017-02-02	en_US
dc.date.issued	2017-04-01	en_US
dc.identifier.citation	Journal of Cell Science, 2017, 130 (7), pp. 1263 - 1273	en_US
dc.identifier.issn	0021-9533	en_US
dc.identifier.uri	http://hdl.handle.net/10453/124842
dc.description.abstract	© 2017. Published by The Company of Biologists Ltd. Members of the Cas family of focal adhesion proteins contain a highly conserved C-terminal focal adhesion targeting (FAT) domain. To determine the role of the FAT domain in these proteins, we compared wild-type exogenous NEDD9 with a hybrid construct in which the NEDD9 FAT domain had been exchanged for the p130Cas (also known as BCAR1) FAT domain. Fluorescence recovery after photobleaching (FRAP) revealed significantly slowed exchange of the fusion protein at focal adhesions and significantly slower twodimensional migration. No differences were detected in cell stiffness as measured using atomic force microscopy (AFM) and in cell adhesion forces measured with a magnetic tweezer device. Thus, the slowed migration was not due to changes in cell stiffness or adhesion strength. Analysis of cell migration on surfaces of increasing rigidity revealed a striking reduction of cell motility in cells expressing the p130Cas FAT domain. The p130Cas FAT domain induced rigiditydependent phosphorylation of tyrosine residues within NEDD9. This in turn reduced post-translational cleavage of NEDD9, which we show inhibits NEDD9-induced migration. Collectively, our data therefore suggest that the p130Cas FAT domain uniquely confers a mechanosensing function.	en_US
dc.relation.ispartof	Journal of Cell Science	en_US
dc.relation.isbasedon	10.1242/jcs.192930	en_US
dc.subject.classification	Developmental Biology	en_US
dc.subject.mesh	Cell Line, Tumor	en_US
dc.subject.mesh	Focal Adhesions	en_US
dc.subject.mesh	Extracellular Matrix	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Tetracycline	en_US
dc.subject.mesh	Adaptor Proteins, Signal Transducing	en_US
dc.subject.mesh	Phosphoproteins	en_US
dc.subject.mesh	Recombinant Fusion Proteins	en_US
dc.subject.mesh	Sequence Alignment	en_US
dc.subject.mesh	Mechanotransduction, Cellular	en_US
dc.subject.mesh	Cell Movement	en_US
dc.subject.mesh	Amino Acid Sequence	en_US
dc.subject.mesh	Structure-Activity Relationship	en_US
dc.subject.mesh	Protein Transport	en_US
dc.subject.mesh	Phosphorylation	en_US
dc.subject.mesh	Crk-Associated Substrate Protein	en_US
dc.subject.mesh	Gene Knockdown Techniques	en_US
dc.subject.mesh	Protein Domains	en_US
dc.title	The focal adhesion targeting domain of p130Cas confers a mechanosensing function	en_US
dc.type	Journal Article
utslib.citation.volume	7	en_US
utslib.citation.volume	130	en_US
utslib.for	0601 Biochemistry and Cell Biology	en_US
utslib.for	1101 Medical Biochemistry and Metabolomics	en_US
utslib.for	06 Biological Sciences	en_US
utslib.for	11 Medical and Health 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	7	en_US
pubs.publication-status	Published	en_US
pubs.volume	130	en_US

Abstract:

© 2017. Published by The Company of Biologists Ltd. Members of the Cas family of focal adhesion proteins contain a highly conserved C-terminal focal adhesion targeting (FAT) domain. To determine the role of the FAT domain in these proteins, we compared wild-type exogenous NEDD9 with a hybrid construct in which the NEDD9 FAT domain had been exchanged for the p130Cas (also known as BCAR1) FAT domain. Fluorescence recovery after photobleaching (FRAP) revealed significantly slowed exchange of the fusion protein at focal adhesions and significantly slower twodimensional migration. No differences were detected in cell stiffness as measured using atomic force microscopy (AFM) and in cell adhesion forces measured with a magnetic tweezer device. Thus, the slowed migration was not due to changes in cell stiffness or adhesion strength. Analysis of cell migration on surfaces of increasing rigidity revealed a striking reduction of cell motility in cells expressing the p130Cas FAT domain. The p130Cas FAT domain induced rigiditydependent phosphorylation of tyrosine residues within NEDD9. This in turn reduced post-translational cleavage of NEDD9, which we show inhibits NEDD9-induced migration. Collectively, our data therefore suggest that the p130Cas FAT domain uniquely confers a mechanosensing function.

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http://hdl.handle.net/10453/124842