SIRT2 与细胞坏死课件.pptVIP

Necrosis What’s the difference between necrosis and apoptosis ? 背景1 肿瘤坏死因子TNF-a通过刺激RIP1和RIP3形成complex，进而磷酸化下游蛋白MLKL。MLKL将“necrosome”与线粒体磷酸酶PGAM5联系起来。PGAM5的激活可以导致成串排列的线粒体发生线性断裂，这一现象在细胞坏死发生的早期起到了非常重要的作用。 Necrosis or Necroptosis Necrostatin-1 背景2 What’s the molecular mechanism that regulates the interaction of RIP1 and RIP3? Sirt2 interacts with RIP3 IP results show that sirt2 interacts with RIP3, and the interaction was unchanged by the addition of the necrotic stimulus TNF-a and z-VAD-fmk over-expressed endogenous Sirt2 interacts with RIP3 Sirt2 regulates RIP1-RIP3 complex formation Mouse embryonic fibroblasts (MEFs) T/z/c:cycloheximide/TNF-a/z-VAD-fmk RIP3 RIP1 Sirt2 RIP1 RIP3 Sirt2 RIP3 RIP1 Sirt2 1 2 Hypothesis sirt2 might modulate the overall necrotic response Absence of a detectable interaction between Sirt2 and RIP1 L929 cells were transfected with an empty vector (-) or with a vector encoding Flag-tagged Sirt2 (+) to assess potential interactions between Sirt2 and RIP1 Sirt2 knockdown blocks RIP1-RIP3 complex formation L292 cell line Inhibition of SIRT2 prevents cellular necrosis Knock down assay shows that sirt2 is an essential factor in necrosis L292 cell line Inhibition of SIRT2 prevents necrosis by blocking RIP1 and RIP3 interaction Treatment with the sirt2 inhibitor AGK2 blocks RIP1-RIP3 complex formation a requirement for SIRT2 in the formation of the RIP1–RIP3 complex summary Copy from Wikipedia Copy from Wikipedia SIRT2 regulates RIP1 acetylation SIRT2 regulates RIP1 acetylation In vitro deacetylase assay Assessment of endogenous RIP1 acetylation and necrosis