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
Purpose:This paper aims to test the use of e-Lattes to map the Brazilian scientific output in a recent research health subject:Zika Virus.Design/methodology/approach:From a set of Lattes CVs of Zika researchers regist...Purpose:This paper aims to test the use of e-Lattes to map the Brazilian scientific output in a recent research health subject:Zika Virus.Design/methodology/approach:From a set of Lattes CVs of Zika researchers registered on the Lattes Platform,we used the e-Lattes to map the Brazilian scientific response to the Zika crisis.Findings:Brazilian science articulated quickly during the public health emergency of international concern(PHEIC)due to the creation of mechanisms to streamline funding of scientific research.Research limitations:We did not assess any dimension of research quality,including the scientific impact and societal value.Practical implications:e-Lattes can provide useful guidelines for different stakeholders in research groups from Lattes CVs of members.Originality/value:The information included in Lattes CVs permits us to assess science from a broader perspective taking into account not only scientific research production but also the training of human resources and scientific collaboration.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Purpose:This paper aims to test the use of e-Lattes to map the Brazilian scientific output in a recent research health subject:Zika Virus.Design/methodology/approach:From a set of Lattes CVs of Zika researchers registered on the Lattes Platform,we used the e-Lattes to map the Brazilian scientific response to the Zika crisis.Findings:Brazilian science articulated quickly during the public health emergency of international concern(PHEIC)due to the creation of mechanisms to streamline funding of scientific research.Research limitations:We did not assess any dimension of research quality,including the scientific impact and societal value.Practical implications:e-Lattes can provide useful guidelines for different stakeholders in research groups from Lattes CVs of members.Originality/value:The information included in Lattes CVs permits us to assess science from a broader perspective taking into account not only scientific research production but also the training of human resources and scientific collaboration.
