Direct cleavage of the human DNA fragmentation factor-45 by granzyme B induces caspase-activated DNase release and DNA fragmentation

被引:104
作者
Sharif-Askari, E
Alam, A
Rhéaume, E
Beresford, PJ
Scotto, C
Sharma, K
Lee, D
DeWolf, WE
Nuttall, ME
Lieberman, J
Sékaly, RP
机构
[1] Univ Montreal, Dept Microbiol & Immunol, Immunol Lab, Montreal, PQ H3C 3J7, Canada
[2] McGill Univ, Dept Microbiol & Immunol, Montreal, PQ H3A 2B4, Canada
[3] McGill Univ, Dept Expt Med, Montreal, PQ H3A 2B4, Canada
[4] Harvard Univ, Sch Med, Ctr Blood Res, Boston, MA 02115 USA
[5] Harvard Univ, Sch Med, Dept Pediat, Boston, MA 02115 USA
[6] SmithKline Beecham Pharmaceut, Dept Med Chem, King Of Prussia, PA 19406 USA
[7] SmithKline Beecham Pharmaceut, Dept Mechanist Enzymol, King Of Prussia, PA 19406 USA
[8] SmithKline Beecham Pharmaceut, Dept Bone & Cartilage Biol, King Of Prussia, PA 19406 USA
关键词
caspace; CTLs; DFF45; granzyme B;
D O I
10.1093/emboj/20.12.3101
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
The protease granzyme B (GrB) plays a key role in the cytocidal activity during cytotoxic T lymphocyte (CTL)-mediated programmed cell death. Multiple caspases have been identified as direct substrates for GrB, suggesting that the activation of caspases constitutes an important event during CTL-induced cell death. However, recent studies have provided evidence for caspase-independent pathway(s) during CTL-mediated apoptosis, In this study, we demonstrate caspase-independent and direct cleavage of the 45 kDa unit of DNA fragmentation factor (DFF45) by GrB both in vitro and in vivo. Using a novel and selective caspase-3 inhibitor, we show the ability of GrB to process DFF45 directly and mediate DNA fragmentation in the absence of caspase-3 activity. Furthermore, studies with DFF45 mutants reveal that both caspase-3 and GrB share a common cleavage site, which is necessary and sufficient to induce DNA fragmentation in target cells during apoptosis. Together, our data suggest that CTLs possess alternative mechanism(s) for inducing DNA fragmentation without the requirement for caspases.
引用
收藏
页码:3101 / 3113
页数:13
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