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dc.contributor.authorTravers, Timothy
dc.contributor.authorShao, Hanshuang
dc.contributor.authorJoughin, Brian A.
dc.contributor.authorLauffenburger, Douglas A.
dc.contributor.authorWells, Alan
dc.contributor.authorCamacho, Carlos J.
dc.date.accessioned2015-10-29T14:07:26Z
dc.date.available2015-10-29T14:07:26Z
dc.date.issued2015-05
dc.date.submitted2014-10
dc.identifier.issn1945-0877
dc.identifier.issn1937-9145
dc.identifier.urihttp://hdl.handle.net/1721.1/99497
dc.description.abstractPhosphorylated residues occur preferentially in the intrinsically disordered regions of eukaryotic proteins. In the disordered amino-terminal region of human a-actinin-4 (ACTN4), Tyr[superscript 4] and Tyr[superscript 31] are phosphorylated in cells stimulated with epidermal growth factor (EGF), and a mutant with phosphorylation-mimicking mutations of both tyrosines exhibits reduced interaction with actin in vitro. Cleavage of ACTN4 by m-calpain, a protease that in motile cells is predominantly activated at the rear, removes the Tyr[superscript 4] site. We found that introducing a phosphomimetic mutation at only Tyr[superscript 31] was sufficient to inhibit the interaction with actin in vitro. However, molecular dynamics simulations predicted that Tyr[superscript 31] is mostly buried and that phosphorylation of Tyr[superscript 4] would increase the solvent exposure and thus kinase accessibility of Tyr[superscript 31]. In fibroblast cells, EGF stimulation increased tyrosine phosphorylation of a mutant form of ACTN4 with a phosphorylation-mimicking residue at Tyr[superscript 4], whereas a truncated mutant representing the product of m-calpain cleavage exhibited EGF-stimulated tyrosine phosphorylation at a background amount similar to that observed for a double phosphomimetic mutant of Tyr[superscript 4] and Tyr[superscript 31]. We also found that inhibition of the receptor tyrosine kinases of the TAM family, such as AXL, blocked EGF-stimulated tyrosine phosphorylation of ACTN4. Mathematical modeling predicted that the kinetics of phosphorylation at Tyr[superscript 31] can be dictated by the kinase affinity for Tyr[superscript 4]. This study suggests that tandem-site phosphorylation within intrinsically disordered regions provides a mechanism for a site to function as a switch to reveal a nearby function-regulating site.en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (Grant R01 GM69668)en_US
dc.language.isoen_US
dc.publisherAmerican Association for the Advancement of Science (AAAS)en_US
dc.relation.isversionofhttp://dx.doi.org/10.1126/scisignal.aaa1977en_US
dc.rightsCreative Commons Attribution-Noncommercial-Share Alikeen_US
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/en_US
dc.sourcePMCen_US
dc.titleTandem phosphorylation within an intrinsically disordered region regulates ACTN4 functionen_US
dc.typeArticleen_US
dc.identifier.citationTravers, T., H. Shao, B. A. Joughin, D. A. Lauffenburger, A. Wells, and C. J. Camacho. “Tandem Phosphorylation Within an Intrinsically Disordered Region Regulates ACTN4 Function.” Science Signaling 8, no. 378 (May 26, 2015): ra51–ra51.en_US
dc.contributor.departmentDavid H. Koch Institute for Integrative Cancer Research at MITen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biological Engineeringen_US
dc.contributor.mitauthorJoughin, Brian A.en_US
dc.contributor.mitauthorLauffenburger, Douglas A.en_US
dc.relation.journalScience Signalingen_US
dc.eprint.versionAuthor's final manuscripten_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dspace.orderedauthorsTravers, T.; Shao, H.; Joughin, B. A.; Lauffenburger, D. A.; Wells, A.; Camacho, C. J.en_US
mit.licenseOPEN_ACCESS_POLICYen_US


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