Ebola virus(EBOV)and Marburg virus(MARV),members of the Filoviridae family,are highly pathogenic and can cause hemorrhagic fevers,significantly impacting human society.Bats are considered reservoirs of these viruses b...Ebola virus(EBOV)and Marburg virus(MARV),members of the Filoviridae family,are highly pathogenic and can cause hemorrhagic fevers,significantly impacting human society.Bats are considered reservoirs of these viruses because related filoviruses have been discovered in bats.However,due to the requirement for maximum containment laboratories when studying infectious viruses,the characterization of bat filoviruses often relies on pseudoviruses and minigenome systems.In this study,we used RACE technology to sequence the 30-leader and 50-trailer of Mengla virus(MLAV)and constructed a minigenome.Similar to MARV,the transcription activities of the MLAV minigenome are independent of VP30.We further assessed the effects of polymorphisms at the 50 end on MLAV minigenome activity and identified certain mutations that decrease minigenome reporter efficiency,probably due to alterations in the RNA secondary structure.The reporter activity upon recombination of the 30-leaders and 50-trailers of MLAV,MARV,and EBOV with those of the homologous or heterologous minigenomes was compared and it was found that the polymerase complex and leader and trailer sequences exhibit intrinsic specificities.Additionally,we investigated whether the polymerase complex proteins from EBOV and MARV support MLAV minigenome RNA synthesis and found that the homologous system is more efficient than the heterologous system.Remdesivir efficiently inhibited MLAV as well as EBOV replication.In summary,this study provides new information on bat filoviruses and the minigenome will be a useful tool for high-throughput antiviral drug screening.展开更多
基金supported by the Key project of the Chinese Academy of Sciences(KJZD-SW-L11 to Z.-L.S.)the Guangzhou Laboratory(SRPG22-001 to Z.-L.S.)+3 种基金the Advanced Customer Cultivation Project of Wuhan National Biosafety Laboratory,Chinese Academy of Sciences(2021ACCP-MS02)the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB0490000 to X.-L.Y.)the Young Topnotch Talent Cultivation Program of Hubei Province(X.-L.Y.)the Youth Innovation Promotion Association of the Chinese Academy of Science(X.-L.Y.).
文摘Ebola virus(EBOV)and Marburg virus(MARV),members of the Filoviridae family,are highly pathogenic and can cause hemorrhagic fevers,significantly impacting human society.Bats are considered reservoirs of these viruses because related filoviruses have been discovered in bats.However,due to the requirement for maximum containment laboratories when studying infectious viruses,the characterization of bat filoviruses often relies on pseudoviruses and minigenome systems.In this study,we used RACE technology to sequence the 30-leader and 50-trailer of Mengla virus(MLAV)and constructed a minigenome.Similar to MARV,the transcription activities of the MLAV minigenome are independent of VP30.We further assessed the effects of polymorphisms at the 50 end on MLAV minigenome activity and identified certain mutations that decrease minigenome reporter efficiency,probably due to alterations in the RNA secondary structure.The reporter activity upon recombination of the 30-leaders and 50-trailers of MLAV,MARV,and EBOV with those of the homologous or heterologous minigenomes was compared and it was found that the polymerase complex and leader and trailer sequences exhibit intrinsic specificities.Additionally,we investigated whether the polymerase complex proteins from EBOV and MARV support MLAV minigenome RNA synthesis and found that the homologous system is more efficient than the heterologous system.Remdesivir efficiently inhibited MLAV as well as EBOV replication.In summary,this study provides new information on bat filoviruses and the minigenome will be a useful tool for high-throughput antiviral drug screening.
