Phosphorylation of FOXO3a on Ser-7 by p38 promotes its nuclear localization in response to doxorubicin.

Ho, Ka-Kei; McGuire, Victoria A; Koo, Chuay-Yeng; Muir, Kyle W; de Olano, Natalia; Maifoshie, Evie; Kelly, Douglas J; McGovern, Ursula B; Monteiro, Lara J; Gomes, Ana R; Nebreda, Angel R; Campbell, David G; Arthur, J Simon C; Lam, Eric W-F

Ho, Ka-Kei; McGuire, Victoria A; Koo, Chuay-Yeng; Muir, Kyle W; de Olano, Natalia; Maifoshie, Evie; Kelly, Douglas J; McGovern, Ursula B; Monteiro, Lara J; Gomes, Ana R; Nebreda, Angel R; Campbell, David G; Arthur, J Simon C; Lam, Eric W-F.

Afiliación

Ho KK; Department of Surgery and Cancer, Imperial College London, Hammersmith Hospital Campus, London W12 0NN, United Kingdom.

J Biol Chem ; 287(2): 1545-55, 2012 Jan 06.

Article en En | MEDLINE | ID: mdl-22128155

RESUMEN

FOXO3a is a forkhead transcription factor that regulates a multitude of important cellular processes, including proliferation, apoptosis, differentiation, and metabolism. Doxorubicin treatment of MCF-7 breast carcinoma cells results in FOXO3a nuclear relocation and the induction of the stress-activated kinase p38 MAPK. Here, we studied the potential regulation of FOXO3a by p38 in response to doxorubicin. Co-immunoprecipitation studies in MCF-7 cells demonstrated a direct interaction between p38 and FOXO3a. We also showed that p38 can bind and phosphorylate a recombinant FOXO3a directly in vitro. HPLC-coupled phosphopeptide mapping and mass spectrometric analyses identified serine 7 as a major site for p38 phosphorylation. Using a phosphorylated Ser-7 FOXO3a antibody, we demonstrated that FOXO3a is phosphorylated on Ser-7 in response to doxorubicin. Immunofluorescence staining studies showed that upon doxorubicin treatment, the wild-type FOXO3a relocalized to the nucleus, whereas the phosphorylation-defective FOXO3a (Ala-7) mutant remained largely in the cytoplasm. Treatment with SB202190 also inhibits the doxorubicin-induced FOXO3a Ser-7 phosphorylation and nuclear accumulation in MCF-7 cells. In addition, doxorubicin caused the nuclear translocation of FOXO3a in wild-type but not p38-depleted mouse fibroblasts. Together, our results suggest that p38 phosphorylation of FOXO3a on Ser-7 is essential for its nuclear relocalization in response to doxorubicin.

Asunto(s)

Antibióticos Antineoplásicos/farmacología; Núcleo Celular/metabolismo; Doxorrubicina/farmacología; Factores de Transcripción Forkhead/metabolismo; Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo; Transporte Activo de Núcleo Celular/efectos de los fármacos; Transporte Activo de Núcleo Celular/genética; Sustitución de Aminoácidos; Animales; Línea Celular Tumoral; Núcleo Celular/genética; Inhibidores Enzimáticos/farmacología; Proteína Forkhead Box O3; Factores de Transcripción Forkhead/genética; Células HEK293; Humanos; Imidazoles/farmacología; Ratones; Ratones Noqueados; Mutación Missense; Fosforilación/efectos de los fármacos; Fosforilación/genética; Piridinas/farmacología; Proteínas Quinasas p38 Activadas por Mitógenos/antagonistas & inhibidores; Proteínas Quinasas p38 Activadas por Mitógenos/genética

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Doxorrubicina / Núcleo Celular / Proteínas Quinasas p38 Activadas por Mitógenos / Factores de Transcripción Forkhead / Antibióticos Antineoplásicos Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: J Biol Chem Año: 2012 Tipo del documento: Article País de afiliación: Reino Unido Pais de publicación: Estados Unidos

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