1. Academic Validation
  2. RIP3-mediated necroptosis is regulated by inter-filament assembly of RIP homotypic interaction motif

RIP3-mediated necroptosis is regulated by inter-filament assembly of RIP homotypic interaction motif

  • Cell Death Differ. 2021 Jan;28(1):251-266. doi: 10.1038/s41418-020-0598-9.
Hong Hu  # 1 2 3 Xialian Wu  # 1 2 3 Guoxiang Wu 1 3 Ning Nan 1 3 Jing Zhang 1 3 Xinxin Zhu 1 2 3 Yu Zhang 1 2 3 Zhaoqian Shu 1 3 Jia Liu 1 3 Xiaoyan Liu 1 Junxia Lu 4 Huayi Wang 5
Affiliations

Affiliations

  • 1 School of Life Science and Technology, ShanghaiTech University, 393 Middle Huaxia Road, Pudong, Shanghai, 201210, China.
  • 2 Chinese Academy of Sciences Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, 320 Yueyang Road, Shanghai, 200031, China.
  • 3 University of Chinese Academy of Sciences, 100864, Beijing, China.
  • 4 School of Life Science and Technology, ShanghaiTech University, 393 Middle Huaxia Road, Pudong, Shanghai, 201210, China. lujx@shanghaitech.edu.cn.
  • 5 School of Life Science and Technology, ShanghaiTech University, 393 Middle Huaxia Road, Pudong, Shanghai, 201210, China. wanghuayi@shanghaitech.edu.cn.
  • # Contributed equally.
Abstract

Necroptosis is mediated by signaling complexes called necrosomes, which contain receptor-interacting protein 3 (RIP3) and upstream effectors, such as RIP1. In necrosomes, the RIP homotypic interaction motif (RHIM) of RIP3 and RIP1 forms amyloidal complex. But how the amyloidal necrosomes control RIP3 activation and cell Necroptosis has not been determined. Here, we showed that RIP3 amyloid fibrils could further assemble into large fibrillar networks which presents as cellular puncta during Necroptosis. A viral RHIM-containing Necroptosis Inhibitor M45 could form heteroamyloid with RIP3 in cells and prevent RIP3 puncta formation and cell Necroptosis. We characterized mutual antagonism between RIP3-RHIM and M45-RHIM in Necroptosis regulation, which was caused by distinct inter-filament interactions in RIP3, M45 amyloids revealed with atomic force microscopy. Moreover, double mutations Asn464 and Met468 in RIP3-RHIM to Asp disrupted RIP3 kinase-dependent Necroptosis. While the mutant RIP3(N464D/M468D) could form amyloid as wild type upon Necroptosis induction. Based on these results, we propose that RIP3 amyloid formation is required but not sufficient in Necroptosis signaling, the ordered inter-filament assembly of RIP3 is critical in RIP3 amyloid mediated kinase activation and cell Necroptosis.

Figures
Products