Tyrosine Phosphorylation of the Myosin Regulatory Light Chain Controls Non-muscle Myosin II Assembly and Function in Migrating Cells
| Autor: | Marina Garrido-Casado, Vanessa C. Talayero, Sarah M. Heissler, María Millán-Salanova, Donald F. Hunt, Miguel Vicente-Manzanares, Clara Llorente-González, Cristina Delgado-Arevalo, Jessica R. Chapman, Jeffrey Shabanowitz, James R. Sellers, Rocío Aguilar-Cuenca |
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| Přispěvatelé: | National Institutes of Health (US), Junta de Castilla y León, Ministerio de Economía y Competitividad (España), Agencia Estatal de Investigación (España), Ministerio de Ciencia, Innovación y Universidades (España), European Commission, Fundación Ramón Areces, Fundación BBVA |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Myosin Light Chains Cell division Myosin CHO Cells macromolecular substances Spodoptera Biology Article General Biochemistry Genetics and Molecular Biology Focal adhesion 03 medical and health sciences chemistry.chemical_compound Cricetulus 0302 clinical medicine Cell Movement Sf9 Cells Animals Humans Phosphorylation Cytoskeleton Migration Actin Myosin Type II Myosin filament Tyrosine phosphorylation Cell biology HEK293 Cells 030104 developmental biology chemistry A549 Cells Tyrosine General Agricultural and Biological Sciences actin 030217 neurology & neurosurgery |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname Curr Biol |
| ISSN: | 0960-9822 2014-5470 |
| Popis: | Active non-muscle myosin II (NMII) enables migratory cell polarization and controls dynamic cellular processes, such as focal adhesion formation and turnover and cell division. Filament assembly and force generation depend on NMII activation through the phosphorylation of Ser19 of the regulatory light chain (RLC). Here, we identify amino acid Tyr (Y) 155 of the RLC as a novel regulatory site that spatially controls NMII function. We show that Y155 is phosphorylated in vitro by the Tyr kinase domain of epidermal growth factor (EGF) receptor. In cells, phosphorylation of Y155, or its phospho-mimetic mutation (Glu), prevents the interaction of RLC with the myosin heavy chain (MHCII) to form functional NMII units. Conversely, Y155 mutation to a structurally similar but non-phosphorylatable amino acid (Phe) restores the more dynamic cellular functions of NMII, such as myosin filament formation and nascent adhesion assembly, but not those requiring stable actomyosin bundles, e.g., focal adhesion elongation or migratory front-back polarization. In live cells, phospho-Y155 RLC is prominently featured in protrusions, where it prevents NMII assembly. Our data indicate that Y155 phosphorylation constitutes a novel regulatory mechanism that contributes to the compartmentalization of NMII assembly and function in live cells. C.L.-G. is supported by a predoctoral fellowship from the Junta de Castilla y León. M.M.-S. is supported by a predoctoral fellowhip from the AECC. This work was funded by the following grants: Programa de Apoyo a Planes Estratégicos de Investigación de Estructuras de Investigación de Excelencia (CLC–2017–01) from the Junta de Castilla-León with FEDER funds (Spain); SAF2014-54705-R and SAF2017-87408-R from MINECO (Spain); CIVP16A1831 from the Ramón Areces Foundation (Spain); 14-BBM-340 from the BBVA Foundation (Spain); and IDEAS-VICE18 from the Asociacion Española Contra el Cáncer (AECC, Spain) to M.V.-M.; NIH GM 037537 (D.F.H.); and K22HL131869 (S.M.H.) and the Intramural Research Program (J.R.S.) of the National Heart, Lung, and Blood Institute, NIH. |
| Databáze: | OpenAIRE |
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