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A direct linkage between the phosphoinositide 3-kinase-AKT signaling pathway and the mammalian target of rapamycin in mitogen-stimulated and transformed cells.
Sekulic, A; Hudson, C C; Homme, J L; Yin, P; Otterness, D M; Karnitz, L M; Abraham, R T.
Afiliación
  • Sekulic A; Department of Immunology, Mayo Clinic, Rochester, Minnesota 55905, USA.
Cancer Res ; 60(13): 3504-13, 2000 Jul 01.
Article en En | MEDLINE | ID: mdl-10910062
The microbially derived antiproliferative agent rapamycin inhibits cell growth by interfering with the signaling functions of the mammalian target of rapamycin (mTOR). In this study, we demonstrate that interleukin-3 stimulation induces a wortmannin-sensitive increase in mTOR kinase activity in a myeloid progenitor cell line. The involvement of phosphoinositide 3'-kinase (PI3K) in the regulation of mTOR activity was further suggested by findings that mTOR was phosphorylated in vitro and in vivo by the PI3K-regulated protein kinase, AKT/PKB. Although AKT phosphorylated mTOR at two COOH-terminal sites (Thr2446 and Ser2448) in vitro, Ser2448 was the major phosphorylation site in insulin-stimulated or -activated AKT-expressing human embryonic kidney cells. Transient transfection assays with mTOR mutants bearing Ala substitutions at Ser2448 and/or Thr2446 indicated that AKT-dependent mTOR phosphorylation was not essential for either PHAS-I phosphorylation or p70S6K activation in HEK cells. However, a deletion of amino acids 2430-2450 in mTOR, which includes the potential AKT phosphorylation sites, significantly increased both the basal protein kinase activity and in vivo signaling functions of mTOR. These results demonstrate that mTOR is a direct target of the PI3K-AKT signaling pathway in mitogen-stimulated cells, and that the identified AKT phosphorylation sites are nested within a "repressor domain" that negatively regulates the catalytic activity of mTOR. Furthermore, the activation status of the PI3K-AKT pathway in cancer cells may be an important determinant of cellular sensitivity to the cytostatic effect of rapamycin.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Quinasas / Transducción de Señal / Proteínas Proto-Oncogénicas / Proteínas Serina-Treonina Quinasas / Fosfotransferasas (Aceptor de Grupo Alcohol) / Fosfatidilinositol 3-Quinasas / Sirolimus Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: Cancer Res Año: 2000 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Quinasas / Transducción de Señal / Proteínas Proto-Oncogénicas / Proteínas Serina-Treonina Quinasas / Fosfotransferasas (Aceptor de Grupo Alcohol) / Fosfatidilinositol 3-Quinasas / Sirolimus Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: Cancer Res Año: 2000 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos