STING signalling is terminated through ESCRT-dependent microautophagy of vesicles originating from recycling endosomes

STING炎症シグナルの終結分子機構 --新規細胞内分解システムの発見--. 京都大学プレスリリース. 2023-03-14.Stimulator of interferon genes (STING) is essential for the type I interferon response against a variety of DNA pathogens. Upon emergence of cytosolic DNA, STING translocates from the endoplasmic reticulum to the Golgi where STING activates the downstream kinase TBK1, then to lysosome through recycling endosomes (REs) for its degradation. Although the molecular machinery of STING activation is extensively studied and defined, the one underlying STING degradation and inactivation has not yet been fully elucidated. Here we show that STING is degraded by the endosomal sorting complexes required for transport (ESCRT)-driven microautophagy. Airyscan super-resolution microscopy and correlative light/electron microscopy suggest that STING-positive vesicles of an RE origin are directly encapsulated into Lamp1-positive compartments. Screening of mammalian Vps genes, the yeast homologues of which regulate Golgi-to-vacuole transport, shows that ESCRT proteins are essential for the STING encapsulation into Lamp1-positive compartments. Knockdown of Tsg101 and Vps4, components of ESCRT, results in the accumulation of STING vesicles in the cytosol, leading to the sustained type I interferon response. Knockdown of Tsg101 in human primary T cells leads to an increase the expression of interferon-stimulated genes. STING undergoes K63-linked ubiquitination at lysine 288 during its transit through the Golgi/REs, and this ubiquitination is required for STING degradation. Our results reveal a molecular mechanism that prevents hyperactivation of innate immune signalling, which operates at REs

Newly Developed Mg2+–Selective Fluorescent Probe Enables Visualization of Mg2+ Dynamics in Mitochondria

Mg2+ plays important roles in numerous cellular functions. Mitochondria take part in intracellular Mg2+ regulation and the Mg2+ concentration in mitochondria affects the synthesis of ATP. However, there are few methods to observe Mg2+ in mitochondria in intact cells. Here, we have developed a novel Mg2+–selective fluorescent probe, KMG-301, that is functional in mitochondria. This probe changes its fluorescence properties solely depending on the Mg2+ concentration in mitochondria under physiologically normal conditions. Simultaneous measurements using this probe together with a probe for cytosolic Mg2+, KMG-104, enabled us to compare the dynamics of Mg2+ in the cytosol and in mitochondria. With this method, carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone (FCCP)–induced Mg2+ mobilization from mitochondria to the cytosol was visualized. Although a FCCP–induced decrease in the Mg2+ concentration in mitochondria and an increase in the cytosol were observed both in differentiated PC12 cells and in hippocampal neurons, the time-courses of concentration changes varied with cell type. Moreover, the relationship between mitochondrial Mg2+ and Parkinson's disease was analyzed in a cellular model of Parkinson's disease by using the 1-methyl-4-phenylpyridinium ion (MPP+). A gradual decrease in the Mg2+ concentration in mitochondria was observed in response to MPP+ in differentiated PC12 cells. These results indicate that KMG-301 is useful for investigating Mg2+ dynamics in mitochondria. All animal procedures to obtain neurons from Wistar rats were approved by the ethical committee of Keio University (permit number is 09106-(1))

Photocatalytic bactericidal action of fluorescent light in a titanium dioxide particle mixture: an in vitro study.

Traditional titanium dioxide (TiO(2)) has photocatalytic bactericidal properties only under ultraviolet (UV) irradiation, which restricts its use in clinical treatment regimens. In this study, we evaluated the photocatalytic bactericidal effects of an aqueous system of TiO(2) particles irradiated by fluorescent light (FL) on Staphylococcus aureus. A TiO(2) particle mixture containing 19 ppm (0.019 mg/mL) of TiO(2) was prepared. A bacterial solution of 1 x 10(5) CFU/mL was added one drop at a time to the TiO(2) mixture. The resulting product was then irradiated with FL. The bacterial survival rate decreased steadily in the TiO(2) mixture group, reaching 76.7% after 30 min of FL irradiation and 10.9% after 60 min. After 60 to 180 min, the bacterial survival ratio of the TiO(2) mixture group was significantly lower than that of the control group (P < 0.05). The present study indicates that treating the surfaces of surgical devices and the surgical field with a TiO(2) particle mixture can create a nearly sterile environment that can be maintained throughout surgery, even at low luminous intensities

The Bactericidal Efficacy of a Photocatalytic TiO2 Particle Mixture with Oxidizer against Staphylococcus aureus

By proving the bactericidal effects of a low-concentration titanium dioxide (TiO2) particle mixture against Staphylococcus aureus, we hope to ultimately apply a mixture of this type as part of a clinical treatment regimen. A bacterial suspension of S. aureus 1×105 CFU/ml was added dropwise to a TiO2 particle mixture (19 ppm TiO2) and irradiated by ultraviolet (UV) light. The colony-forming units were counted and the bacterial survival rate was calculated. In the control sample, the bacterial survival rate was 83.3% even after 120 min. In the TiO2 mixture + UV sample, the bacteria count dropped sharply, reaching 17.3% of the baseline value at 30 min and 0.4% at 60 min. TiO2 particles dispersed in water mixtures are known to elicit highly efficient UV absorption and greater bonding to bacteria. A reaction of the TiO2 with another oxidizer altered the aqueous pH and accelerated the photocatalytic chemical reaction. The TiO2 particle mixture showed high antibacterial action against S. aureus even at a low concentration

Photocatalytic TiO2 particles confer superior antibacterial effects in a nutrition-rich environment: an in vitro study

Titanium dioxide (TiO2) is known to confer photocatalytic bactericidal effects under ultraviolet (UV) irradiation. Few reports are available, however, on the clinical applications of TiO2 particle mixtures. Our objective in the present research was to evaluate the in vitro bactericidal effects of a TiO2 particle mixture in a nutrition-rich biological environment. A bacterial suspension of Staph-ylococcus aureus and epidermidis 3 × 103CFU/mL was added to a TiO2 particle mixture (0.038 mg/mL) containing mainly sodium percarbonate and citric acid. To simulate a biological environment, 40 μL of 10% bovine serum albumin was added and the culture temperature was maintained at 37°C. The resulting product was irradiated by UV light and the bacterial survival rate was calculated for each time of UV irradiation. In the control sample treated with distilled water + UV, the bacteria survived at a high rate even after 180 min. In the TiO2 mixture+ UV sample, meanwhile, the bacterial survival rate dropped to 43.8% and 6.0% of the baseline values in S. aureus and S. epidermidis, respectively, after 60 min of UV irradiation. The photocatalytic antibacterial action of the TiO2 particle mixture was high even in a protein-rich biological environment