Identification of a novel phosphorylation site, Ser-170, as a regulator of bad pro-apoptotic activity

被引:72
作者
Dramsi, S
Scheid, MP
Maiti, A
Hojabrpour, P
Chen, XM
Schubert, K
Goodlett, DR
Aebersold, R
Duronio, V [1 ]
机构
[1] Univ British Columbia, Dept Med, Vancouver, BC V6H 3Z6, Canada
[2] Vancouver Hosp, Jack Bell Res Ctr, Vancouver, BC V6H 3Z6, Canada
[3] Inst Syst Biol, Seattle, WA 98105 USA
关键词
D O I
10.1074/jbc.M109990200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Bad is a pro-apoptotic member of the Bcl-2 family of proteins that is thought to exert a death-promoting effect by heterodimerization with Bcl-X-L, nullifying its anti-apoptotic activity. Growth factors may promote cell survival at least partially through phosphorylation of Bad at one or more of Ser-112, -136, or -155. Our previous work showed that Bad is also phosphorylated in response to cytokines at another site, which we now identify as Ser-170. The functional role of this novel phosphorylation site was assessed by site-directed mutagenesis and analysis of the pro-apoptotic function of Bad in transiently transfected HEK293 and COS-7 cells or by stable expression in the cytokine-dependent cell line, MC/9. In general, mutation of Ser-170 to Ala results in a protein with increased ability to induce apoptosis, similar to the S112A mutant. Mutation of Ser-170 to Asp, mimicking a constitutively phosphorylated site, results in a protein that is virtually unable to induce apoptosis. Similarly, the S112A/S170D double mutant does not cause apoptosis in HEK293 and MC/9 cell lines. These data strongly suggest that phosphorylation of Bad at Ser-170 is a critical event in blocking the pro-apoptotic activity of Bad.
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收藏
页码:6399 / 6405
页数:7
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