Invasive bacterial pathogens are internalized by host cells through endocytosis, which is regulated by a cascade of actin rearrangement signals triggered by host cell receptors or bacterial proteins ...delivered into host cells. However, the molecular mechanisms that mediate actin rearrangement to promote bacterial invasion are not fully understood. Here, we show that the autophagy-related (ATG) protein ATG9B regulates the internalization of various bacteria by controlling actin rearrangement. ATG knockout screening and knockdown experiments in HeLa cells identified ATG9B as a critical factor for bacterial internalization. In particular, cells with ATG9B knockdown exhibited an accumulation of actin filaments and phosphorylated LIM kinase and cofilin, suggesting that ATG9B is involved in actin depolymerization. Furthermore, the kinase activity of Unc-51-like autophagy-activating kinase 1 was found to regulate ATG9B localization and actin remodeling. These findings revealed a newly discovered function of ATG proteins in bacterial infection rather than autophagy-mediated immunity.
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•ATG9B regulates bacterial internalization via actin rearrangement in Epithelial cells•ATG9B modulates actin rearrangement through the phosphorylation cascade of LIMK/cofilin•ULK1 is involved in bacterial internalization through the regulation of ATG9B localization
Microbiology; Cell biology
This study examined the effects of alloying elements such as Re and Ta on the microstructural evolution of recrystallized W under proton (1 MeV H+) and self-ion (18 MeV W6+) irradiations at 800 °C. ...Although the number density of voids increased with increasing proton-induced damage level, the void density in W-Re (R) and W-Ta (R) alloys were lower than that of pure W (R). Herein, the addition of Re and Ta to W suppresses the void formation process. In the proton-irradiated W-3%Re (R), a lot of dislocation loops were observed at 0.05 dpa which is the stage of nucleation. The evolution process up to 0.2 dpa was characterized by loop growth via the absorption of clusters and point defects. The dislocation loops then coalesce and grow large, and the dislocation lines become tangled at 1 dpa. At 0.05 dpa, the dislocation loops in pure W (R) have already evolved into the tangled dislocations. Solute Re may inhibit the mobility of small dislocation loops and SIA clusters. In W-3%Ta (R) irradiated at 0.05 and 0.2 dpa, the coalescence process of the elongated dislocation loops was observed. Solute Ta may inhibit the mobility of SIA clusters. Although no voids and rafts were observed in self-ion irradiated W-3%Re (R) to 0.2 dpa, not only dislocation loops but also voids and rafts were observed in pure W (R) to 0.2 dpa. Solute Re may inhibit the mobility of small dislocation loops and SIA clusters, so that solute Re would suppress the raft formation and then the void formation under self-ion irradiation. The dispersed barrier model was applied to the irradiation hardening of the proton-irradiated W-3%Re (R) up to 0.2 dpa before developing dislocation lines. The hardening in the proton-irradiated W-3%Re (R) up to 0.2 dpa is mainly due to TEM-visible voids and dislocation loops.
Macroautophagy/autophagy plays an important role in the immune response to invasion by intracellular pathogens such as group A Streptococcus (GAS; Streptococcus pyogenes). We previously identified ...RAB30, a Golgi-resident GTPase, as a novel anti-bacterial autophagic regulator in the formation of GAS-containing autophagosome-like vacuoles (GcAVs); however, the precise mechanism underlying this process remains elusive. Here, we elucidate a novel property of RAB30: the ability to recruit PI4KB (phosphatidylinositol 4-kinase beta) to the Golgi apparatus and GcAVs. We found that trans-Golgi network (TGN) vesicles were incorporated into GcAVs via RAB30 to promote GcAV formation. Moreover, depletion of phosphatidylinositol-4-phosphate (PtdIns4P), a phosphatidylinositol enriched in the TGN, by wortmannin and phenylarsine oxide, followed by subsequent repletion with exogenous PtdIns4P revealed that PtdIns4P is crucial for GcAV formation. Furthermore, we identify an interaction between RAB30 and PI4KB, in which the knockdown of RAB30 decreased the localization of PI4KB to the TGN and GcAVs. Finally, PI4KB knockout suppressed autophagy by inhibiting GcAV formation, resulting in the increased survival of GAS. Our results demonstrate a novel autophagosomal formation mechanism involving coordinative functions of RAB30 and PI4KB distinct from those utilized in canonical autophagy.
Abbreviations: GAS: group A Streptococcus; GcAVs: GAS-containing autophagosome-like vacuoles; PI4KB: phosphatidylinositol 4-kinase beta; PtdIns: phosphatidylinositol; PtdIns3P: phosphatidylinositol-3-phosphate; PtdIns4P: phosphatidylinositol-4-phosphate; PtdIns5P: phosphatidylinositol-5-phosphate; SLO: streptolysin O; TGN: trans-Golgi network; TGOLN2: trans-golgi network protein 2; PH: plekstrin homology; OSBP: oxysterol binding protein
Bacterial autophagy-a type of macroautophagy that is also termed xenophagy-selectively targets intracellular bacteria such as group A
(GAS), a ubiquitous pathogen that causes numerous serious ...diseases, including pharyngitis, skin infections, and invasive life-threatening infections. Although bacterial autophagy is known to eliminate invading bacteria
the action of autophagy receptors, the underlying mechanism remains unclear. Herein, we elucidated that Tollip functions as a bacterial-autophagy receptor in addition to participating involved in the intracellular immunity mechanism that defends against bacterial infection. Tollip was recruited to GAS-containing endosomal vacuoles prior to the escape of GAS into the cytosol; additionally, Tollip knockout disrupted the recruitment of other autophagy receptors, such as NBR1, TAX1BP1, and NDP52, to GAS-containing autophagosomes and led to prolonged intracellular survival of GAS. Furthermore, Tollip was found to be required for the recruitment of galectin-1 and -7 to GAS-containing autophagosomes, and immunoprecipitation results indicated that Tollip interacts with galectin-7. Lastly, our data also revealed that galectin-1 and -7 are involved in the restriction of GAS replication in cells. These results demonstrated that Tollip modulates bacterial autophagy by recruiting other autophagy receptors and galectins.
Autophagy acts as a host-defense system against pathogenic microorganisms such as Group A Streptococcus (GAS). Autophagy is a membrane-mediated degradation system that is regulated by intracellular ...membrane trafficking regulators, including small GTPase Rab proteins. Here, we identified Rab30 as a novel regulator of GAS-containing autophagosome-like vacuoles (GcAVs). We found that Rab30, a Golgi-resident Rab, was recruited to GcAVs in response to autophagy induction by GAS infection in epithelial cells. Rab30 recruitment was dependent upon its GTPase activity. In addition, the knockdown of Rab30 expression significantly reduced GcAV formation efficiency and impaired intracellular GAS degradation. Rab30 normally functions to maintain the structural integrity of the Golgi complex, but GcAV formation occurred even when the Golgi apparatus was disrupted. Although Rab30 also colocalized with a starvation-induced autophagosome, Rab30 was not required for autophagosome formation during starvation. These results suggest that Rab30 mediates autophagy against GAS independently of its normal cellular role in the structural maintenance of the Golgi apparatus, and autophagosome biogenesis during bacterial infection involves specific Rab GTPases.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Anti-apoptotic Bcl-2 and Bcl-xL are proposed to regulate starvation-induced autophagy by directly interacting with Beclin 1. Beclin 1 is also thought to be involved in multiple vesicle trafficking ...pathways such as endocytosis by binding to Atg14L and UVRAG. However, how the interaction of Bcl-2 family proteins and Beclin 1 regulates anti-bacterial autophagy (xenophagy) is still unclear. In this study, we analyzed these interactions using Group A Streptococcus (GAS; Streptococcus pyogenes) infection as a model. GAS is internalized into epithelial cells through endocytosis, while the intracellular fate of GAS is degradation by autophagy. Here, we found that Bcl-xL but not Bcl-2 regulates GAS-induced autophagy. Autophagosome-lysosome fusion and the internalization process during GAS infection were promoted in Bcl-xL knockout cells. In addition, knockout of Beclin 1 phenocopied the internalization defect of GAS. Furthermore, UVRAG interacts not only with Beclin 1 but also with Bcl-xL, and overexpression of UVRAG partially rescued the internalization defect of Beclin 1 knockout cells during GAS infection. Thus, our results indicate that Bcl-xL inhibits GAS-induced autophagy directly by suppressing autophagosome-lysosome fusion and indirectly by suppressing GAS internalization via interaction with Beclin 1-UVRAG.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The purpose of the present study is to clarify the instability behavior of M23C6 under irradiation, specifically the occurrence of radiation-induced amorphization (RIA). Ion irradiation of 10.5 ...MeV-Fe3+ at elevated temperatures from 573 to 623 K was conducted into the reduced activation ferritic/martensitic steels (F82H) and its model alloy (Fe-8Cr-0.1C). A bilayer contrast of the particle consisting of an amorphous-rim phase and inner crystalline core of M23C6 was observed in the irradiated F82H specimen, but not in the model alloy. From the high-resolution electron microscope observation, the preferential occupation site of W into M23C6 lattice was identified as 8c-site prior to irradiation in F82H specimen, which shifted to other sites due to chemical disordering upon irradiation. Evaluation of the intensity ratio between 8c and another site of M23C6, 8c/4a, then revealed that the extent of chemical disordering of W was mitigated at the amorphous-crystal interface region in comparison with the central of the particle. The hypothesis for the formation mechanism of an amorphous-rim in M23C6 was presumed as the deviation from the stoichiometric composition at the local interface due to the irradiation-enhanced diffusion and/or ballistic mixing under the current circumstances, although the efforts from experimental and/or simulation studies are still necessary to achieve a further understanding of the RIA behavior in M23C6.
Size and temperature dependence of the point defect binding free energy has numerically evaluated for self-interstitial atom (SIA) clusters and vacancy clusters in bcc Fe by using continuum models ...based on thermodynamics and linear elasticity. The estimated binding free energy of SIAs to SIA-clusters is much higher than that of vacancies to vacancy clusters, indicating that SIA-clusters are more thermally stable than vacancy clusters. For relatively small clusters, the estimated binding free energy at 0 K is comparably consistent with atomistic calculation data; and then, the SIA binding free energy at 850 K is averagely about 35% lower than that at 0 K, while the vacancy binding free energy is about 6% lower; which may remarkably affect the formation kinetics of those defect clusters under irradiation. These kinds of information will be one of the basic parameters for a theoretical model of the microstructural evolution of Fe-based materials in the nuclear fusion DEMO environment.
Group A
(GAS;
) is a major human pathogen that causes streptococcal pharyngitis, skin and soft tissue infections, and life-threatening conditions such as streptococcal toxic-shock syndrome. During ...infection, GAS not only invades diverse host cells but also injects effector proteins such as NAD-glycohydrolase (Nga) into the host cells through a streptolysin O (SLO)-dependent mechanism without invading the cells; Nga and SLO are two major virulence factors that are associated with increased bacterial virulence. Here, we have shown that the invading GAS induces fragmentation of the Golgi complex and inhibits anterograde transport in the infected host cells through the secreted toxins SLO and Nga. GAS infection-induced Golgi fragmentation required both bacterial invasion and SLO-mediated Nga translocation into the host cytosol. The cellular Golgi network is critical for the sorting of surface molecules and is thus essential for the integrity of the epithelial barrier and for the immune response of macrophages to pathogens. In epithelial cells, inhibition of anterograde trafficking by invading GAS and Nga resulted in the redistribution of E-cadherin to the cytosol and an increase in bacterial translocation across the epithelial barrier. Moreover, in macrophages, interleukin-8 secretion in response to GAS infection was found to be suppressed by intracellular GAS and Nga. Our findings reveal a previously undescribed bacterial invasion-dependent function of Nga as well as a previously unrecognized GAS-host interaction that is associated with GAS pathogenesis.
Two prominent virulence factors of group A
(GAS), streptolysin O (SLO) and NAD-glycohydrolase (Nga), are linked to enhanced pathogenicity of the prevalent GAS strains. Recent advances show that SLO and Nga are important for intracellular survival of GAS in epithelial cells and macrophages. Here, we found that invading GAS disrupts the Golgi complex in host cells through SLO and Nga. We show that GAS-induced Golgi fragmentation requires bacterial invasion into host cells, SLO pore formation activity, and Nga NADase activity. GAS-induced Golgi fragmentation results in the impairment of the epithelial barrier and chemokine secretion in macrophages. This immune inhibition property of SLO and Nga by intracellular GAS indicates that the invasion of GAS is associated with virulence exerted by SLO and Nga.
Liquid metals are excellent coolants of fission and fusion reactors. However, the chemical compatibility of structural materials is important issue. The mass losses of the high-temperature materials ...such as FeCrAl-ODS, SiC, and refractory metals by corrosion in liquid metals are essential information to obtain their corrosion rates. The specimens must be cleaned to remove liquid metals solidified and adhered on the specimens after the corrosion tests, though the damage of the specimens in the cleaning procedure must be minimized. Cleaning methodologies appropriate for the specimens tested in liquid metals are urgently required for further compatibility study. The cleaning methodology with 0.1 M sodium hydroxide (NaOH) solution was developed, in which Sn was selectively dissolved without any damage on the specimens of the high-temperature materials. The cleaning procedure to remove Pb, Bi, and these alloys (i.e., Pb-16Li and Pb-Bi) adhered on the specimens in the solution mixture of acetic acid, ethanol, and H2O2 were also studied.
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