PARP5A and RNF146 phase separation restrains RIPK1-dependent necroptosis

Shouqiao Hou; Jian Zhang; Xiaoyan Jiang; Yuanxin Yang; Bing Shan; Mengmeng Zhang; Cong Liu; Junying Yuan; Daichao Xu

doi:10.1016/j.molcel.2023.12.041

PARP5A and RNF146 phase separation restrains RIPK1-dependent necroptosis

Mol Cell. 2024 Mar 7;84(5):938-954.e8. doi: 10.1016/j.molcel.2023.12.041. Epub 2024 Jan 24.

Authors

Shouqiao Hou¹, Jian Zhang², Xiaoyan Jiang³, Yuanxin Yang¹, Bing Shan³, Mengmeng Zhang³, Cong Liu⁴, Junying Yuan⁵, Daichao Xu⁶

Affiliations

¹ Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 201210, China; University of Chinese Academy of Sciences, Beijing 101408, China.
² Department of Neurosurgery, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215031, China.
³ Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 201210, China.
⁴ Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 201210, China; Shanghai Key Laboratory of Aging Studies, Shanghai 201210, China; State Key Laboratory of Chemical Biology, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China.
⁵ Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 201210, China; Shanghai Key Laboratory of Aging Studies, Shanghai 201210, China.
⁶ Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 201210, China; Shanghai Key Laboratory of Aging Studies, Shanghai 201210, China; State Key Laboratory of Chemical Biology, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China. Electronic address: xudaichao@sioc.ac.cn.

PMID: 38272024
DOI: 10.1016/j.molcel.2023.12.041

Abstract

Phase separation is a vital mechanism that mediates the formation of biomolecular condensates and their functions. Necroptosis is a lytic form of programmed cell death mediated by RIPK1, RIPK3, and MLKL downstream of TNFR1 and has been implicated in mediating many human diseases. However, whether necroptosis is regulated by phase separation is not yet known. Here, we show that upon the induction of necroptosis and recruitment by the adaptor protein TAX1BP1, PARP5A and its binding partner RNF146 form liquid-like condensates by multivalent interactions to perform poly ADP-ribosylation (PARylation) and PARylation-dependent ubiquitination (PARdU) of activated RIPK1 in mouse embryonic fibroblasts. We show that PARdU predominantly occurs on the K376 residue of mouse RIPK1, which promotes proteasomal degradation of kinase-activated RIPK1 to restrain necroptosis. Our data demonstrate that PARdU on K376 of mouse RIPK1 provides an alternative cell death checkpoint mediated by phase separation-dependent control of necroptosis by PARP5A and RNF146.

Keywords: PARP5A; PARylation; RIPK1; RNF146; necroptosis; phase separation; ubiquitination.

MeSH terms

Animals
Apoptosis / physiology
Cell Death
Fibroblasts / metabolism
Mice
Necroptosis* / genetics
Phase Separation*
Receptor-Interacting Protein Serine-Threonine Kinases / genetics
Receptor-Interacting Protein Serine-Threonine Kinases / metabolism
Ubiquitin-Protein Ligases / genetics
Ubiquitin-Protein Ligases / metabolism

Substances

Receptor-Interacting Protein Serine-Threonine Kinases
Ubiquitin-Protein Ligases
Rnf146 protein, mouse
Ripk1 protein, mouse