Mosquito-borne flaviviruses,such as Zika virus(ZIKV)and dengue virus(DENV),cause diverse severe clinical manifestations including fever,rash,hepatitis,arthralgia,and congenital anomalies.Here,we identified a host fact...Mosquito-borne flaviviruses,such as Zika virus(ZIKV)and dengue virus(DENV),cause diverse severe clinical manifestations including fever,rash,hepatitis,arthralgia,and congenital anomalies.Here,we identified a host factor,the adaptor protein complex 1 gamma 1 subunit(AP1G1),which plays an important role in both ZIKV and dengue virus 2(DENV2)infections.We explored the role of AP1G1 in ZIKV and DENV2 infections using CRISPR/Cas9 gene editing technology and RNA interference(RNAi)techniques.Knockout or silencing of AP1G1 decreases the replication of ZIKV and DENV2 in multiple human cell lines.Intriguingly,depletion of AP1G1 results in a significant reduction in ZIKV at an early stage,but decreases DENV2 replication levels during the late stage,suggesting that AP1G1 plays distinct roles in the infection by ZIKV and DENV2.Furthermore,we determined that AP1G1 mediates ZIKV-endosomal membrane fusion through inhibitor experiments and fluorescence labeling assays.Mechanistically,we found that AP1G1 exerts its pro-viral effect through binding to the ZIKV envelope glycoprotein(E protein).This interaction promotes the fusion of viral and endosomal membranes,during which the ZIKV genomic RNAs are released from the endosome into the cytoplasm,a process that facilitates viral replication.However,for DENV2 infection,AP1G1 primarily affects its viral RNA replication stage,rather than the fusion of virus-endosomal membrane.Taken together,our work demonstrates that AP1G1 plays a pro-viral role in both ZIKV and DENV2 infections via distinct mechanisms,highlighting its potential as a therapeutic target for antiviral strategies.展开更多
The flower’s meristematic characteristic gene AP1 was introduced into Chrysanthemum morifolium cv. ‘Yu Ren Mian’ mediated by Agrobacterium tumefaciens.The factors influencing genetic transformation protocol were st...The flower’s meristematic characteristic gene AP1 was introduced into Chrysanthemum morifolium cv. ‘Yu Ren Mian’ mediated by Agrobacterium tumefaciens.The factors influencing genetic transformation protocol were studied.The results showed that the leaf explants precultured for 2～8 hours or not precultured were best for transformation by A. tumefaciens. The suitable concentration of bacterial and the time for infecting was OD_ 600 0.5 for 10 minutes. Leaves were cocultivated with bacterial at 23～25 ℃ for 2 days,then delayed selection for 3 days. Kan^r plant were selected by increased selective press from 5 mg·L ~ -1 G418 to 7.5 mg5L~ -1 G418 followed by 10 mg5L~ -1 G418. The integration of AP1 gene into C. morifolium ‘Yu Ren Mian’ was confirmed by PCR and Southern blotting.Two of transgenic plants bloomed 15 days earlier than untransformed plants.展开更多
基金supported by the National Natural Science Foundation of China(No.82471389,No.32470986,No.82271385)Natural Science Foundation of Guangdong Province(No.2024A1515010471).
文摘Mosquito-borne flaviviruses,such as Zika virus(ZIKV)and dengue virus(DENV),cause diverse severe clinical manifestations including fever,rash,hepatitis,arthralgia,and congenital anomalies.Here,we identified a host factor,the adaptor protein complex 1 gamma 1 subunit(AP1G1),which plays an important role in both ZIKV and dengue virus 2(DENV2)infections.We explored the role of AP1G1 in ZIKV and DENV2 infections using CRISPR/Cas9 gene editing technology and RNA interference(RNAi)techniques.Knockout or silencing of AP1G1 decreases the replication of ZIKV and DENV2 in multiple human cell lines.Intriguingly,depletion of AP1G1 results in a significant reduction in ZIKV at an early stage,but decreases DENV2 replication levels during the late stage,suggesting that AP1G1 plays distinct roles in the infection by ZIKV and DENV2.Furthermore,we determined that AP1G1 mediates ZIKV-endosomal membrane fusion through inhibitor experiments and fluorescence labeling assays.Mechanistically,we found that AP1G1 exerts its pro-viral effect through binding to the ZIKV envelope glycoprotein(E protein).This interaction promotes the fusion of viral and endosomal membranes,during which the ZIKV genomic RNAs are released from the endosome into the cytoplasm,a process that facilitates viral replication.However,for DENV2 infection,AP1G1 primarily affects its viral RNA replication stage,rather than the fusion of virus-endosomal membrane.Taken together,our work demonstrates that AP1G1 plays a pro-viral role in both ZIKV and DENV2 infections via distinct mechanisms,highlighting its potential as a therapeutic target for antiviral strategies.
文摘The flower’s meristematic characteristic gene AP1 was introduced into Chrysanthemum morifolium cv. ‘Yu Ren Mian’ mediated by Agrobacterium tumefaciens.The factors influencing genetic transformation protocol were studied.The results showed that the leaf explants precultured for 2～8 hours or not precultured were best for transformation by A. tumefaciens. The suitable concentration of bacterial and the time for infecting was OD_ 600 0.5 for 10 minutes. Leaves were cocultivated with bacterial at 23～25 ℃ for 2 days,then delayed selection for 3 days. Kan^r plant were selected by increased selective press from 5 mg·L ~ -1 G418 to 7.5 mg5L~ -1 G418 followed by 10 mg5L~ -1 G418. The integration of AP1 gene into C. morifolium ‘Yu Ren Mian’ was confirmed by PCR and Southern blotting.Two of transgenic plants bloomed 15 days earlier than untransformed plants.
