摘要
旨在研究宿主热休克蛋白HSP90AB1对猪繁殖与呼吸综合征病毒(PRRSV)复制的影响。选用对PRRSV易感的MARC-145细胞系,探究过表达和干扰HSP90AB1对病毒复制的影响,利用激光共聚焦免疫荧光和免疫共沉淀验证HSP90AB1与PRRSV的非结构蛋白NSP2作用关系,利用双荧光素酶试验探究HSP90AB1和NSP2对NF-kB活性的影响。结果表明,过表达HSP90AB1后显著抑制病毒的复制,干扰HSP90AB1的表达后促进病毒的复制;免疫共沉淀结果和激光共聚焦免疫荧光说明,HSP90AB1和NSP2存在相互作用;双荧光素酶报告试验表明HSP90AB1能逆转NSP2对NF-κB活性的抑制作用。综上,HSP90AB1可能通过与PRRSV NSP2相互作用,拮抗NSP2对NF-kB活性的抑制来抑制病毒的复制和感染。
In order to study the effect of overexpression and interference of host heat shock protein HSP90 AB1 on the replication of porcine reproductive and respiratory syndrome virus(PRRSV),and explore its mechanism.The MARC-145 cell line susceptible to PRRSV was selected,and the effect of overexpression and interference of HSP90 AB1 on viral replication was investigated.Secondly,laser confocal immunofluorescence and immunoprecipitation were used to verify the relationship between HSP90 AB1 and non structural protein NSP2 of PRRSV.Double luciferase experiment was used to explore the effect of HSP90 AB1 and NSP2 on NF-κB activity.The results showed that after overexpression of HSP90 AB1 and overexpression of HSP90 AB1,the replication of the virus was significantly inhibited.After interfering with the expression of HSP90 AB1,the replication of the virus was promoted.The results of immunoprecipitation and laser confocal immunofluorescence showed that HSP90 AB1 interacted with NSP2.The double luciferase report experiment showed that HSP90 AB1 could reverse the effect of NSP2 on NF-κB activity inhibition.In conclusion,HSP90 AB1 may inhibit virus replication and infection by antagonizing the inhibition of NF-κB activity by NSP2 through PRRSV NSP2 interaction.
作者
谢丰玉
康磊
陈梦莉
郑家杨
李宗杰
李蓓蓓
邵东华
邱亚峰
魏建超
马志永
刘珂
黎满香
XIE Fengyu;KANG Lei;CHEN Mengli;ZHENG Jiayang;LI Zongjie;LI Beibei;SHAO Donghua;QIU Yafeng;WEI Jianchao;MA Zhiyong;LIU Ke;LI Manxiang(College of Veterinary Medicine,Hunan Agricultural University,Changsha 410128,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China)
出处
《畜牧与兽医》
CAS
北大核心
2022年第12期128-135,共8页
Animal Husbandry & Veterinary Medicine
基金
上海市自然科学基金面上项目(21ZR1477100)。
