Ubiquitin-like protein MNSFβ/endophilin II complex regulates Dectin-1-mediated phagocytosis and inflammatory responses in macrophages

Post-translational modification by monoclonal nonspecific suppressor factor β (MNSFβ) has been implicated in the regulation of a variety of cellular events. Previous studies have demonstrated that MNSFβ covalently binds to the intracellular pro-apoptotic protein Bcl-G in a macrophage cell line, Raw264.7, suggesting involvement of this ubiquitin-like protein in apoptosis. Most recently, we found that MNSFβ covalently conjugates to endophilin II, a member of the endophilin A family, and inhibits phagocytosis by macrophages. In this study, we further examined the mechanism of action of MNSFβ/endophilin II complex in the phagocytosis of zymosan. MNSFβ/endophilin II I mediated inhibition of phagocytosis in Raw264.7 cells was neutralized by anti-Decti-1, β-glucan receptor, mAb, indicating that MNSFβ/endophilin II is a mediator of Dectin-1 signaling in regulating phagocytosis. The β-glucan-dependent TNFα response to zymosan was significantly increased by the treatment with endophilin II siRNA and/or MNSFβ siRNA. Conversely, cotransfection of endophilin II and MNSFβ cDNAs inhibited the enhancement of zymosan-induced TNFα production. Interestingly, endophilin II siRNA did not affect Pam₃CSK₄ (TLR2 specific ligand)-induced TNFα production. Endophilin II and/or MNSFβ siRNA enhanced zymosan-induced IκBα degradation. Together, these results demonstrate that MNSFβ/endophilin II inhibits the signal pathway upstream of IKK activation, but not downstream of TLR2 signaling.