目的探讨IQSEC1是否通过病毒蛋白PB1调控甲型流感病毒的增殖。方法首先克隆甲型流感病毒[A/Shanghai/02/2013(H7N9)]的8个基因;其次,通过免疫共沉淀检测IQ模体Sec7结构域蛋白1(IQSEC1)与聚合酶PB1(PB1)存在相互作用;此外,通过过表达或...目的探讨IQSEC1是否通过病毒蛋白PB1调控甲型流感病毒的增殖。方法首先克隆甲型流感病毒[A/Shanghai/02/2013(H7N9)]的8个基因;其次,通过免疫共沉淀检测IQ模体Sec7结构域蛋白1(IQSEC1)与聚合酶PB1(PB1)存在相互作用;此外,通过过表达或者敲低IQSEC1的方法检测IQSEC1对PB1核定位的影响;最后,过表达或者敲低IQSEC1后检测Influenza A virus[A/Shanghai/02/2013(H7N9)]。结果病毒感染条件下,外源IQSEC1和PB1存在相互作用。当过表达IQSEC1时,细胞中IQSEC1的表达量上升,相应的PB1在细胞核中的定位减少;当用敲低IQSEC1时,细胞中IQSEC1的表达量下降,相应的PB1在细胞核中的定位上升。过表达IQSEC1后,甲型流感病毒的增殖水平下降(P<0.05)。敲低IQSEC1后,甲型流感病毒的增殖水平上升(P<0.05)。结论IQSEC1通过减少甲型流感病毒蛋白PB1的核定位抑制甲型流感病毒的增殖。展开更多
The general secretory(Sec)pathway represents a common mechanism by which bacteria secrete proteins,including virulence factors,into the extracytoplasmic milieu.However,there is little information about this system,as ...The general secretory(Sec)pathway represents a common mechanism by which bacteria secrete proteins,including virulence factors,into the extracytoplasmic milieu.However,there is little information about this system,as well as its associated secretory proteins,in relation to the fire blight pathogen Erwinia amylovora.In this study,data mining revealed that E.amylovora harbors all of the essential components of the Sec system.Based on this information,we identified putative Sec-dependent secretory proteases in E.amylovora on a genome-wide scale.Using the programs SignalP,LipoP,and Phobius,a total of 15 putative proteases were predicted to contain the N-terminal signal peptides(SPs)that might link them to the Sec-dependent pathway.The activities of the predicted SPs were further ttvreaaalicsdyeta otgepelda nsuesmsi inicng cprarenoa psEeesrtdcy h.s eiTrgirncaihfniicsa carcniotplltiyi-obwnahasle enad nEaa.ll kyaaslmiensye lo spvhohorowas epwdh atasht aaust sete h(deP theoo xipAnr)o ecgsueslinaotene f iuofms i1 om1 na tosufyr steth epe em1 a t5 rh s,ae txs tcuroagcngfyeitrsomtipnelagd s ttmhhieecii rr pperoox---xL?tential roles in plant infection.The results of this study support the suggestion that E.amylovora might employ the Sec system to secrete a suite of proteases to enable successful infection of plants,and shed new light on the interaction of E.amylovora with host plants.展开更多
文摘目的探讨IQSEC1是否通过病毒蛋白PB1调控甲型流感病毒的增殖。方法首先克隆甲型流感病毒[A/Shanghai/02/2013(H7N9)]的8个基因;其次,通过免疫共沉淀检测IQ模体Sec7结构域蛋白1(IQSEC1)与聚合酶PB1(PB1)存在相互作用;此外,通过过表达或者敲低IQSEC1的方法检测IQSEC1对PB1核定位的影响;最后,过表达或者敲低IQSEC1后检测Influenza A virus[A/Shanghai/02/2013(H7N9)]。结果病毒感染条件下,外源IQSEC1和PB1存在相互作用。当过表达IQSEC1时,细胞中IQSEC1的表达量上升,相应的PB1在细胞核中的定位减少;当用敲低IQSEC1时,细胞中IQSEC1的表达量下降,相应的PB1在细胞核中的定位上升。过表达IQSEC1后,甲型流感病毒的增殖水平下降(P<0.05)。敲低IQSEC1后,甲型流感病毒的增殖水平上升(P<0.05)。结论IQSEC1通过减少甲型流感病毒蛋白PB1的核定位抑制甲型流感病毒的增殖。
基金Project supported by the Key Technologies R&D Program of Xinjiang Production and Construction Corps(No.2018AB038)the National Natural Science Foundation of China(No.31970126)。
文摘The general secretory(Sec)pathway represents a common mechanism by which bacteria secrete proteins,including virulence factors,into the extracytoplasmic milieu.However,there is little information about this system,as well as its associated secretory proteins,in relation to the fire blight pathogen Erwinia amylovora.In this study,data mining revealed that E.amylovora harbors all of the essential components of the Sec system.Based on this information,we identified putative Sec-dependent secretory proteases in E.amylovora on a genome-wide scale.Using the programs SignalP,LipoP,and Phobius,a total of 15 putative proteases were predicted to contain the N-terminal signal peptides(SPs)that might link them to the Sec-dependent pathway.The activities of the predicted SPs were further ttvreaaalicsdyeta otgepelda nsuesmsi inicng cprarenoa psEeesrtdcy h.s eiTrgirncaihfniicsa carcniotplltiyi-obwnahasle enad nEaa.ll kyaaslmiensye lo spvhohorowas epwdh atasht aaust sete h(deP theoo xipAnr)o ecgsueslinaotene f iuofms i1 om1 na tosufyr steth epe em1 a t5 rh s,ae txs tcuroagcngfyeitrsomtipnelagd s ttmhhieecii rr pperoox---xL?tential roles in plant infection.The results of this study support the suggestion that E.amylovora might employ the Sec system to secrete a suite of proteases to enable successful infection of plants,and shed new light on the interaction of E.amylovora with host plants.
