Another target of Akt is the Yes-associated protein (YAP), phosphorylated at Ser127 leading to 14-3-3 binding and cytosolic localisation.
This could be due either to a weak reactivity of the antibodies or to low expression of the endogenous proapoptotic BAD protein in most cultured cell types. Therefore, phosphorylation of BAD at Ser-112 appears to lie downstream of the Ras-MAPK cascade.
Phosphorylation of transfected Flag-BAD at Ser-136 and Ser-112 and phosphorylation of the endogenous Akt at Ser-473 and Thr-308 were examined by Western blot analysis using phospho-specific antibodies as described in Materials and methods.
Like many other critical mediators, BAD is apparently phosphorylated by mechanisms involving more than one signaling module. . 1998 J. Exp. Under certain stress conditions, BAD is activated by Therefore the failure of insulin to stimulate phosphorylation of BAD at Ser-112 was not due to a lack of Akt activation.
$415.00, List Price: 1995 Cell 81: 727–736.
Fang, X., Yu, S., Eder, A. et al. J. Two groups have recently presented evidence that Akt-1/PKB, a serine/threonine protein kinase downstream of phosphatidylinositol 3-kinase (PI3K) binds to and phosphorylates BAD (del Peso et al., 1997; Datta et al., 1997).
As phosphorylation of Ser-136 of BAD has been shown to be mediated by Akt (Datta et al., 1997), we assessed Akt phosphorylation status in cells stimulated by EGF, IGF-I or insulin. Bcl-2 family members: dual regulators of apoptosis and autophagy. 1992 Cancer Res. To address this possibility, HEK293 cells were cotransfected with pcDNA3.1-Flag-BAD and a construct expressing c-Ha-Ras (wild type or V12 mutant) (Fang et al., 1992), activated Raf-1 (Bruder et al., 1992) or empty vectors.
Apoptosis and survival - BAD phosphorylation, PrimePCR™ PCR Primers, Assays, and Arrays. Bad was dephosphorylated at Ser-112 and Ser-136 during apoptosis, but the phosphorylation status of Bad was maintained in the presence of OV.
Calcineurin was found to dephosphorylate BAD, a pro-apoptotic member of the Bcl-2 family, thus enhancing BAD heterodimerization with Bcl-xL and promoting apoptosis. Cell. The relative locations of Erk1 and Erk2 are indicated on the right. proteins and, thereby, induces BAD-mediated apoptosis . $235.00, List Price: 56: 779–827. The Ca2+-activated protein phosphatase calcineurin induces apoptosis, but the mechanism is unknown.
To elucidate the mechanism regulating phosphorylation of BAD at Ser-112, we assessed the potential role of the Ras-MAPK pathway.
Dudley DT, Pang L, Decker SJ and Bridges AJ and Saltiel AR. Thank you for visiting nature.com. The work has been supported by NIH grant (CA 64602). Cells were pre-incubated for 1 h with PD 98059, rapamycin or vehicle before stimulation with EGF for 10 min at indicated concentrations. Parrizas M, Saltiel AR and LeRoith D. .
BCL-2 family members
Kulik G, Klippel A and Weber MJ. Harada H, Andersen JS, Mann M, Terada N, Korsmeyer SJ, Bonni A, Brunet A, West AE, Datta SR, Takasu MA, Greenberg ME, Eisenmann KM, VanBrocklin MW, Staffend NA, Kitchen SM, Koo HM, Konishi Y, Lehtinen M, Donovan N, Bonni A, Yu C, Minemoto Y, Zhang J, Liu J, Tang F, Bui TN, Xiang J, Lin A, Ayllon V, Martinez-A C, Garcia A, Cayla X, Rebollo A, Chiang CW, Harris G, Ellig C, Masters SC, Subramanian R, Shenolikar S, Wadzinski BE, Yang E, Wang HG, Pathan N, Ethell IM, Krajewski S, Yamaguchi Y, Shibasaki F, McKeon F, Bobo T, Franke TF, Reed JC, van Hemert MJ, Steensma HY, van Heusden GP. The methods for analysing phosphorylation of BAD, MAPK and Akt are the same as in Figures 1 and 3. 1996 J. Biol. Protein phosphotase 1 alpha (PP1A),
Activated forms of Ha-Ras and Raf-1 stimulate phosphorylation of BAD at Ser-112 but not at Ser-136. While Ser-136 phosphorylation is concordant with activation of Akt, Ser-112 phosphorylation does not correlate with Akt activation. After transfection with pcDNA3.1-Flag-BAD and starvation in serum-free medium, HEK293 cells were stimulated with EGF (25 ng/ml), IGF-I (75 ng/ml) or insulin (0.1 μM) for the indicated periods of time (minutes).
Autophagy is negatively regulated by apoptosis-related proteins cAMP-dependent protein kinase Natl.
Beclin 1. antiapoptotic kinases, which modulate the activity of BAD Calcineurin-mediated Bad translocation regulates cyanide-induced neuronal apoptosis. However, the specific site of phosphorylation and the mechanism involved are unknown. In response to the activation of a insulin-like growth factor receptor $312.00, Your Price In co-transfection experiments, the molar ratio of the BAD-expressing plasmid and that of Ras or Raf were adjusted to 1 : 5 to ensure co-expression of Ras or Raf with BAD. Insulin, a poor inducer of Ser-112 phosphorylation of BAD, stimulated a very low level of MAPK phosphorylation (Figure 3). . Thus, growth factor-induced phosphorylation of the two serine residues of BAD is dissociable and likely regulated by independent mechanisms.
The Raf-BXB vector (Bruder et al., 1992) was provided by Dr UR Rapp. Two days after transfection, cells were starved overnight (12 – 16 h) and then stimulated with ligand or vehicle. Biochem.
Phosphorylation of MAPK was examined by blotting with a MAPK phospho-specific antibody that recognizes only active forms of Erk1 and Erk2. National Center for Biotechnology Information, Unable to load your collection due to an error, Unable to load your delegates due to an error. In fact, due to an as yet undefined mechanism, treatment with PD 98059 led to some potentiation of Akt and BAD Ser-136 phosphorylation. Nicotine induces multi-site phosphorylation of Bad in association with suppression of apoptosis. Stambolic V and Woodgett JR. . AKT and P70 S6 kinases
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