Heat stress activates RIPK3 through ZBP1

热应激通过ZBP1激活RIPK3

RIPK1, Toll/interleukin-1 receptor domain–containing adapter-inducing interferon-b(TRIF), and ZBP1 are RIP homotypic interaction motif (RHIM)–containing proteins that can interact with and activate RIPK3 (5, 19, 20).

RIPK1,Toll/interleukin-1受体结构域包含适配器可诱导干扰素B，ZBP1和RIP同型相互作用基序蛋白质，可以与RIPK3相互作用并激活他；

We therefore investigated whether these RIPK3- interacting proteins are required for heat stress–induced cell death. Deletion of TRIF, mutation of the kinase domain of RIPK1 (RIPK1D/D), or inhibition of RIPK1 by necrostain-1 did not affect heat stress–induced cell death in BMDMs(Fig. 3A and fig. S6, A and B). 

因此我们研究了这些RIPK3作用蛋白对热应激诱导的细胞死亡是否是必需的。TRIF的缺失，RIPK1激酶域的突变，或者necrostain-1对RIPK1的抑制都不能影响BMDM中热应激诱导的细胞死亡

RIPK1 deficiency did not significantly inhibit heat stress–induced cell death in L929 cells (fig. S6A). 

RIPK1缺乏不能显著抑制L929细胞中热应激诱导的细胞死亡

By contrast,deletion of ZBP1 abrogated the phosphorylation of RIPK3 and MLKL; the cleavage of casp8, casp3, and GSDME; and cell death after heat stress (Fig. 3, A and B). These findings were confirmed in L929 cells (fig. S6, C and D).

相比之下，ZBP1的缺失消除了RIPK3和MLKL的磷酸化，casp8，casp3和GSDME的裂解和热应激后的细胞死亡。这些发现都在L929细胞中被加以证实。

Restoration of ZBP1 expression restored the capacity of Zbp1-deficient cells to undergo cell

death after heat stress (Fig. 3C). Human HT-29, a colon cancer cell line that express RIPK3 and RIPK1 but not ZBP1, also failed to undergo heat stress–induced cell death (Fig.3D). 

恢复ZBP1的表达后，回复了ZBP1-缺失细胞在热应激条件下激活细胞死亡的能力。人类HT-29，人类结肠癌细胞系，可以表达RIPK3和RIPK1，但是不表达ZBP1,也无法在热应激条件下诱导细胞死亡。

Expression of exogenous human ZBP1 rendered HT-29 cells susceptible to heat stress–

induced cell death (Fig. 3D). 

HT-29细胞中外源性ZBP1的表达使细胞对热应激诱导的死亡更加敏感

Furthermore, heat stress induced the interaction between ZBP1 and RIPK3 (Fig. 3, E and F). These data demonstrate that heat stress activates RIPK3 through ZBP1.

另外，热应激诱导了ZBP1和RIPK3的相互作用。这些数据证明，热应激是通过ZBP1来激活RIPK3。

今日没有什么心得

只是我的条带显得不好

难过子

做得不顺利是真的要及时止损