Objectives To identify the 5'untranslated region of Zika virus(ZIKV 5'UTR)RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site(IRES)located...Objectives To identify the 5'untranslated region of Zika virus(ZIKV 5'UTR)RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site(IRES)located in ZIKV 5'UTR and virus production.Methods Interacting proteins in U251 cells were captured using tRSA-tagged ZIKV 5'UTR RNA and tRSA-ZIKV 5'UTR RNA-binding proteins were visualized by SDS-PAGE silver staining,Subsequently,liquid chromatographytandem mass spectrometry(LC-MS/MS),bioinformatics analysis,and Western blot were used to identify the candidate proteins binding to ZIKV 5'UTR.Dicistronic expression assay and plaque forming assay were performed to analyze the effect of the binding protein on ZIKV IRES activity and ZIKV production,respecitvely.Results tRSA RNA pull-down assay,LC-MS/MS,and Western blot analysis showed that polypyrimidine tractbinding protein(PTB)bound to the ZIKV 5'UTR.Furthermore,dual luciferase reporter assay revealed that overexpression of PTB significantly enhanced the IRES activity of ZIKV(t=10.220,P<0.001),while PTB knockdown had the opposite effect(t=4.897,P<0.01).Additionally,virus plaque forming assay demonstrated that up-regulation of PTB expression significantly enhanced viral titer(t=6.400,P<0.01),whereas reducing PTB expression level weakened virus infectivity(t=5.055,P<0.01).Conclusion PTB positively interacts with the ZIKV 5'UTR and enhances IRES activity and virus production.展开更多
The past twenty years have seen the increasingly important role of ontology in traditional Chinese medicine(TCM).However,the development of TCM ontology faces many challenges.Since the epistemologies dramatically diff...The past twenty years have seen the increasingly important role of ontology in traditional Chinese medicine(TCM).However,the development of TCM ontology faces many challenges.Since the epistemologies dramatically differ between TCM and contemporary biomedicine,it is hard to apply the existing top-level ontology mechanically.“Data silos”are widely present in the currently available terminology standards,term sets,and ontologies.The formal representation of ontology needs to be further improved in TCM.Therefore,we propose a unified basic semantic framework of TCM based on in-depth theoretical research on the existing top・level ontology and a re-study of important concepts in TCM.Under such a framework,ontologies in TCM subdomains should be built collaboratively and be represented formally in a common format.Besides,extensive cooperation should be encouraged by establishing ontology research communities to promote ontology peer review and reuse.展开更多
文摘Objectives To identify the 5'untranslated region of Zika virus(ZIKV 5'UTR)RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site(IRES)located in ZIKV 5'UTR and virus production.Methods Interacting proteins in U251 cells were captured using tRSA-tagged ZIKV 5'UTR RNA and tRSA-ZIKV 5'UTR RNA-binding proteins were visualized by SDS-PAGE silver staining,Subsequently,liquid chromatographytandem mass spectrometry(LC-MS/MS),bioinformatics analysis,and Western blot were used to identify the candidate proteins binding to ZIKV 5'UTR.Dicistronic expression assay and plaque forming assay were performed to analyze the effect of the binding protein on ZIKV IRES activity and ZIKV production,respecitvely.Results tRSA RNA pull-down assay,LC-MS/MS,and Western blot analysis showed that polypyrimidine tractbinding protein(PTB)bound to the ZIKV 5'UTR.Furthermore,dual luciferase reporter assay revealed that overexpression of PTB significantly enhanced the IRES activity of ZIKV(t=10.220,P<0.001),while PTB knockdown had the opposite effect(t=4.897,P<0.01).Additionally,virus plaque forming assay demonstrated that up-regulation of PTB expression significantly enhanced viral titer(t=6.400,P<0.01),whereas reducing PTB expression level weakened virus infectivity(t=5.055,P<0.01).Conclusion PTB positively interacts with the ZIKV 5'UTR and enhances IRES activity and virus production.
文摘The past twenty years have seen the increasingly important role of ontology in traditional Chinese medicine(TCM).However,the development of TCM ontology faces many challenges.Since the epistemologies dramatically differ between TCM and contemporary biomedicine,it is hard to apply the existing top-level ontology mechanically.“Data silos”are widely present in the currently available terminology standards,term sets,and ontologies.The formal representation of ontology needs to be further improved in TCM.Therefore,we propose a unified basic semantic framework of TCM based on in-depth theoretical research on the existing top・level ontology and a re-study of important concepts in TCM.Under such a framework,ontologies in TCM subdomains should be built collaboratively and be represented formally in a common format.Besides,extensive cooperation should be encouraged by establishing ontology research communities to promote ontology peer review and reuse.
