E3 ligase TRIM25 ubiquitinates RIP3 to inhibit TNF induced cell necrosis

Abstract

Receptor interacting protein kinase 3 (RIP3 or RIPK3), the critical executor of cell programmed necrosis, plays essential roles in maintaining immune responses and appropriate tissue homeostasis. Although the E3 ligases CHIP and PELI1 are reported to promote RIP3 degradation, however, how post-translational modification regulates RIP3 activity and stability is poorly understood. Here, we identify the tripartite motif protein TRIM25 as a negative regulator of RIP3-dependent necrosis. TRIM25 directly interacts with RIP3 through its SPRY domain and mediates the K48-linked polyubiquitination of RIP3 on residue K501. The RING domain of TRIM25 facilitates the polyubiquitination chain on RIP3, thereby promoting proteasomal degradation of RIP3. Also, TRIM25 deficiency inhibited the ubiquitination of RIP3, thus promoting TNF-induced cell necrosis. Our current finding reveals the regulating mechanism of polyubiquitination on RIP3, which might be a potential therapeutic target for the intervention of RIP3-dependent necrosis-related diseases.

Surviving death: emerging concepts of RIPK3 and MLKL ubiquitination in the regulation of necroptosis

“…The tripartite motif E3 ligase TRIM25 interacts also with RIPK3, but not with RIPK1 or MLKL, and mediates ubiquitination of human RIPK3 at K501 [186]. Although loss of TRIM25 expression sensitised human and murine cell lines towards necroptosis, due to TRIM25-and K48-linked poly-Ub-mediated degradation of RIPK3 by the 26S proteasome, RIPK3 Ub: ubiquitin, RHIM: RIP homotypic interaction motif, 4HB: four-helix bundle, PsKD, pseudokinase domain.…”

The role of Ubiquitination in Apoptosis and Necroptosis

“…Another E3 ligase, TRIM25, was shown to induce degradation of RIPK3 via K48-linked chains [ 109 ]. Mutation of K501 of RIPK3 markedly reduced its TRIM25-dependent ubiquitination and increased TNF-α-induced necroptosis, indicating that TRIM25 is anti-necroptotic by reducing the expression of RIPK3 via proteasomal degradation [ 109 ]. However, overexpression experiments of mutant Ub were used to determine chain types conjugated to RIPK3, so should be interpreted with caution.…”

“… LUBAC* (M1) No effect? [ 101 ] OTULIN** (M1) ↑Necroptosis [ 103 ] CHIP* ↓Necroptosis [ 108 ] CYLD** ↑Necroptosis [ 112 , 113 ] RIPK3 K5 (Ms) A20** (K63) ↓Necroptosis [ 105 ] K55, K363 CHIP* ↓Necroptosis [ 108 ] K363 Pellino-1* (K48) ↓Necroptosis [ 110 ] K197, K302, K364 Parkin* (K33) ↓Necroptosis [ 107 ] K501 TRIM25*(K48) ↓Necroptosis [ 109 ] K518 USP22** ↑Necroptosis [ 106 ] MLKL K219 (Ms) ??? * (K63) ↑Necroptosis [ 119 ] ???…”

The Emerging Role of Deubiquitinases in Cell Death

“…RIPK3 ubiquitination is involved in the regulation of necroptosis. The K5, K42, K55, K197, K302, K351, K364, K363, K469, K501, and K518 sites are critical ubiquitination modification sites [ 66 , 67 , 68 , 69 , 70 , 71 ]. The E3 ligase STIP1 homology and U-box containing protein 1 (STUB1, also known as carboxy terminus of HSP70-interacting protein/CHIP) can regulate cell necroptosis through ubiquitylation- and lysosome-dependent RIPK3 degradation by K55 and K363 ubiquitination [ 66 ].…”

“…The pellino E3 ubiquitin-protein ligase 1 (PELI1) mediates K48-linked polyubiquitylation of RIPK3 on lysine 363, leading to proteasomal degradation of RIPK3 [ 67 ]. The E3 ligase tripartite motif containing 25 (TRIM25) binds to RIPK3, promotes polyubiquitination of K48-linked RIPK3 at residue K501, and negatively regulates RIPK3 stability through the ubiquitin–proteasome degradation pathway [ 68 ]. The E3 ligase Parkin (PRKN) is implicated in RIPK3 ubiquitination at K197, K302, and K364 residues conjugated K33-linked ubiquitin chains during TNF-induced necroptosis [ 69 ].…”