MARCH3 attenuates IL-1β–triggered inflammation by mediating K48-linked polyubiquitination and degradation of IL-1RI

Abstract

The proinflammatory cytokine IL-1β plays critical roles in inflammatory and autoimmune diseases. IL-1β signaling is tightly regulated to avoid excessive inflammatory response. In this study, we identified the E3 ubiquitin ligase membrane-associated RING-CH-type finger 3 (MARCH3) as a critical negative regulator of IL-1β–triggered signaling. Overexpression of MARCH3 inhibited IL-1β–triggered activation of NF-κB as well as expression of inflammatory genes, whereas MARCH3 deficiency had the opposite effects. MARCH3-deficient mice produced higher levels of serum inflammatory cytokines and were more sensitive to inflammatory death upon IL-1β injection or Listeria monocytogenes infection. Mechanistically, MARCH3 was associated with IL-1 receptor I (IL-1RI) and mediated its K48-linked polyubiquitination at K409 and lysosomal-dependent degradation. Furthermore, IL-1β stimulation triggered dephosphorylation of MARCH3 by CDC25A and activation of its E3 ligase activity. Our findings suggest that MARCH3-mediated IL-1RI degradation is an important mechanism for attenuating IL-1β–triggered inflammatory response.

RNF 152 positively regulates TLR / IL ‐1R signaling by enhancing MyD88 oligomerization

“…Recombinant human IL‐1β (Peprotech), peptidoglycan (PGN; Sigma), LPS (Sigma), and SYBR (Bio‐Rad) were purchased from the indicated companies. Monoclonal anti‐Flag M2 antibody and monoclonal anti‐Flag M2‐Peroxidase (HRP) antibody (Sigma‐Aldrich, 1:1,000); antibodies against HA (OriGene, 1:2,000), myc (ABclonal, 1:2,000), β‐actin (ABclonal, 1:10,000), phospho‐IKKα (Ser176)/IKKβ (Ser177; C84E11; Cell Signaling Technology, 1:500), IKKβ (D30C6; Cell Signaling Technology, 1:2,000), phospho‐p38 (Thr180/Tyr182; 28B10; Cell Signaling Technology, 1:500), p38 (Cell Signaling Technology, 1:2,000), GST (91G1; Cell Signaling Technology, 1:2,000), MyD88 (R&D Systems, AF3109, 1:500), IRAK1 (D51G7; Cell Signaling Technology, 1:2,000), TLR4 (ABclonal, A17634, 1:1,000), and mouse antisera against IL1R1 were described previously. APC rat anti‐mouse CD8a (BD Biosciences, 1:200); Pacific Blue™ rat anti‐mouse CD4 (BD Biosciences, 1:200); PE rat anti‐mouse CD4 (BD Biosciences, 1:200); PE rat anti‐mouse CD25 (BD Biosciences, 1:200); APC rat anti‐mouse CD45R/B220 (BD Biosciences, 1:200); FITC rat anti‐mouse CD3 molecular complex (BD Biosciences, 1:500); PE anti‐mouse CD49b antibody (BioLegend, 1:200); mouse IL‐1β ELISA MAX™ Deluxe (BioLegend); LEGEND MAX™ Mouse IFN‐β ELISA Kit (BioLegend); and mouse TNF‐α ELISA MAX™ Deluxe (BioLegend) were purchased from the indicated companies.…”

“…Upon stimulation, MyD88 is recruited to the receptor via TIR-TIR interactions, which in turn releases the DD of MyD88 and consequently initiates its oligomerization [10]. Although intensive efforts have been made to characterize the signaling mediated by TLR/IL-1R [11][12][13][14][15][16][17], the regulation of MyD88 oligomerization is still enigmatic.…”

The Membrane-Associated MARCH E3 Ligase Family: Emerging Roles in Immune Regulation

“…MARCH1 is localized on LAMP-1-positive late endosomes/lysosomes, which is consistent with the finding that MARCH1 mediates ubiquitination and lysosome-dependent degradation of MHC class II molecules (24, 25). MARCH3 and MARCH8 regulate interleukin-1 (IL-1) receptor complex-mediated signaling (26). MARCH4 is located on the Golgi apparatus (15).…”

“…For example, MARCH3 is kept inactive by TYRO3-mediated phosphorylation in unstimulated cells. Upon IL-1β stimulation, CDC25A dephosphorylates MARCH3, which in turn activates MARCH3 and causes K48-linked polyubiquitination and degradation of IL-1 receptor type I (IL-1RI), leading to inhibition of IL-1β-triggered signaling (26).…”

Role of RING-Type E3 Ubiquitin Ligases in Inflammatory Signalling and Inflammatory Bowel Disease

“…Nevertheless, despite the formation of heterodimers of RNF40 with RNF20, RNF40 alone activates NF- κ B signalling and upregulates NF- κ B-dependent transcription by promoting I κ B kinase (IKK) phosphorylation and p65 nuclear translocation, indicating the involvement of NF- κ B-dependent transcription in the ubiquitination of substrates other than H2B [ 34 ]. MARCH3 (RNF173) mediates Lys48-linked ubiquitination and lysosomal degradation of IL-1 receptor I and thereby inhibits IL-1 β -triggered NF- κ B activation [ 35 ]. Independent of its E3 ligase activity, RNF8 inhibits TNF- α -induced NF- κ B activation by directly binding with the kinase domain of IKK and interfering with IKK α / β phosphorylation [ 36 ].…”