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Genesis ; 52(11): 907-15, 2014 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-25242698

RESUMEN

Focal adhesion kinase is a non-receptor protein tyrosine kinase with signaling functions downstream of integrins and growth factor receptors. In addition to its role in adhesion, migration, and proliferation it also has non-kinase scaffolding functions in the nucleus. Focal adhesion kinase (FAK) activation involves the following: (1) ligand bound growth factors or clustered integrins activate FAK kinase domain; (2) FAK autophosphorylates tyrosine (Y) 397; (3) Src binds pY397 and phosphorylates FAK at various other sites including Y861; (4) downstream signaling of activated FAK elicits changes in cellular behavior. Although many studies have demonstrated roles for the kinase domain, Y397 and Y861 sites, in vitro much less is known about their functions in vivo. Here, we report the generation of a series of FAK-mutant knockin mice where mutant FAK, either kinase dead, non-phosphorylatable mutants Y397F and Y861F, or mutant Y397E-containing a phosphomimetic site that results in a constitutive active Y397, can be expressed in a Cre inducible fashion driven by the ROSA26 promoter. In future studies, intercrossing these mice with FAKflox/flox mice and inducible cre-expressing mice will enable the in vivo study of mutant FAK function in the absence of endogenous FAK in a spatially and temporally regulated fashion within the whole organism.


Asunto(s)
Activación Enzimática/fisiología , Quinasa 1 de Adhesión Focal/genética , Modelos Animales , Mutación Puntual/genética , Transducción de Señal/genética , Animales , Secuencia de Bases , Western Blotting , Activación Enzimática/genética , Técnica del Anticuerpo Fluorescente , Técnicas de Sustitución del Gen , Vectores Genéticos/genética , Inmunoprecipitación , Ratones , Sondas Moleculares/genética , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Fosforilación , Reacción en Cadena en Tiempo Real de la Polimerasa , Tamoxifeno
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