With GSK0660 GST-NS1 full or GST-NS1 N but not with GST-NS1 C (Figure 2F).Co-localization of NS1 and b-tubulin in CellsA549 cells were transfected with pCMV5-HA-NS1, NS1 was apparent from 24 h post-transfection, mainly in nucleus (green color) (Figure 3E). On the other hand, b-tubulin was stained in nucleus and cytoplasm (red color) (Figure 3F).The signals of NS1 and b-tubulin clearly overlapped in nucleus (Figure 3G).NS1 Interacts with b-Tubulinmerization, thereby arrest the cell cycle in G2/M phase and disrupt normal cell division, further act through several types of kinases, leading to phosphorylation cascades and the activation of cyclin B1/cdc2 complex and Bcl-2 phosphorylation, finally initiates the apoptotic cascade [39,40,41]. Our 18334597 observation indicated influenza virus A/Beijing/501/2009(H1N1) NS1 caused G2-M cell cycle arrest (data not shown), moreover caspase 3-dependent apoptosis was showed to be involved in the homologous strain A/Wenshan H1N1-induced A549 cell and CNE-2Z cell death. Taken together, we presumed that the interaction of influenza virus A/Beijing/501/ 2009(H1N1) NS1 with b-tubulin depolymerized MT network and thereby disrupt normal cell division and commit the cell to apoptosis, thereby facilitate virus replication and indirectly contribute to virus pathogenicity. However, the exact role ofNS1 on apoptosis induced by the 2009 pandemic H1N1 virus needs further investigation. In summary, the present study provides evidence that b-tubulin represent a novel interaction partner of influenza A virus NS1 protein. The RNA-binding domain of NS1 is responsible for binding with b-tubulin. The interaction of NS1 with b-tubulin disrupts the cellular microtubule network and induces apoptosis on human A549 cells.Author ContributionsConceived and designed the experiments: ZHL XQH BHL XML. Performed the experiments: ZHL XQH HYW LM. Analyzed the data: ZHL XQH SQW BHL XML. Contributed reagents/materials/analysis tools: ZHL HJC LM SQW TYZ. Wrote the paper: ZHL XQH HJC BHL XML.
Pharmacologically active constituents in extracts of the medicinal licorice root include glycyrrhizin (GA) and its aglycone metabolite 18b-glycyrrhetinic acid (GRA). Both compounds have been extensively studied for their effects on cellular physiology and as immune system modulators in cultured cell lines, in small animal models and in humans, with either or both demonstrating anti-tumorgenic, anti-allergenic, anti-hepatotoxic, antiviral, antiulcerative, or anti-inflammatory properties (reviewed in [1]). Multiple mechanisms of activity have been proposed including inductive or inhibitory effects on apoptosis, cytokine expression, intracellular signaling pathways, transcription factor activation, cellular membrane fluidity and modulation of oxidative GSK0660 web stress [1?6]. How or if these mechanisms function in vivo to account for the ability of these compounds to attenuate pathology in infectious and inflammatory diseases is not well understood. GA has been shown to be beneficial in vivo in several systems. In the clinical setting, intravenous administration of a commercial formulation containing GA (Stronger Neo-MinophagenH) has been used in Japan for .20 years to treat patients with chronic viral hepatitis, with evidence of clinical improvement and reduction in progression to hepatocellular carcinoma [7?0]. Murine models of infectious and inflammatory diseases providefurther evidence for immune modulating or antimicrobial properties of GA. GA reduces lethality associated w.With GST-NS1 full or GST-NS1 N but not with GST-NS1 C (Figure 2F).Co-localization of NS1 and b-tubulin in CellsA549 cells were transfected with pCMV5-HA-NS1, NS1 was apparent from 24 h post-transfection, mainly in nucleus (green color) (Figure 3E). On the other hand, b-tubulin was stained in nucleus and cytoplasm (red color) (Figure 3F).The signals of NS1 and b-tubulin clearly overlapped in nucleus (Figure 3G).NS1 Interacts with b-Tubulinmerization, thereby arrest the cell cycle in G2/M phase and disrupt normal cell division, further act through several types of kinases, leading to phosphorylation cascades and the activation of cyclin B1/cdc2 complex and Bcl-2 phosphorylation, finally initiates the apoptotic cascade [39,40,41]. Our 18334597 observation indicated influenza virus A/Beijing/501/2009(H1N1) NS1 caused G2-M cell cycle arrest (data not shown), moreover caspase 3-dependent apoptosis was showed to be involved in the homologous strain A/Wenshan H1N1-induced A549 cell and CNE-2Z cell death. Taken together, we presumed that the interaction of influenza virus A/Beijing/501/ 2009(H1N1) NS1 with b-tubulin depolymerized MT network and thereby disrupt normal cell division and commit the cell to apoptosis, thereby facilitate virus replication and indirectly contribute to virus pathogenicity. However, the exact role ofNS1 on apoptosis induced by the 2009 pandemic H1N1 virus needs further investigation. In summary, the present study provides evidence that b-tubulin represent a novel interaction partner of influenza A virus NS1 protein. The RNA-binding domain of NS1 is responsible for binding with b-tubulin. The interaction of NS1 with b-tubulin disrupts the cellular microtubule network and induces apoptosis on human A549 cells.Author ContributionsConceived and designed the experiments: ZHL XQH BHL XML. Performed the experiments: ZHL XQH HYW LM. Analyzed the data: ZHL XQH SQW BHL XML. Contributed reagents/materials/analysis tools: ZHL HJC LM SQW TYZ. Wrote the paper: ZHL XQH HJC BHL XML.
Pharmacologically active constituents in extracts of the medicinal licorice root include glycyrrhizin (GA) and its aglycone metabolite 18b-glycyrrhetinic acid (GRA). Both compounds have been extensively studied for their effects on cellular physiology and as immune system modulators in cultured cell lines, in small animal models and in humans, with either or both demonstrating anti-tumorgenic, anti-allergenic, anti-hepatotoxic, antiviral, antiulcerative, or anti-inflammatory properties (reviewed in [1]). Multiple mechanisms of activity have been proposed including inductive or inhibitory effects on apoptosis, cytokine expression, intracellular signaling pathways, transcription factor activation, cellular membrane fluidity and modulation of oxidative stress [1?6]. How or if these mechanisms function in vivo to account for the ability of these compounds to attenuate pathology in infectious and inflammatory diseases is not well understood. GA has been shown to be beneficial in vivo in several systems. In the clinical setting, intravenous administration of a commercial formulation containing GA (Stronger Neo-MinophagenH) has been used in Japan for .20 years to treat patients with chronic viral hepatitis, with evidence of clinical improvement and reduction in progression to hepatocellular carcinoma [7?0]. Murine models of infectious and inflammatory diseases providefurther evidence for immune modulating or antimicrobial properties of GA. GA reduces lethality associated w.
