Investigating the role of ANP32A in the replication of Avian Influenza Virus

流感病毒感染家禽和猪等家畜，造成收入损失、社会和经济混乱。禽流感仍然是粮食安全的主要威胁之一。禽流感在动物宿主中的适应性导致病毒在人类中传播，并有可能引发紧急流感大流行。我们研究了一个家族的宿主蛋白，ANP 32，这是增选流感病毒，以支持其复制。鸡和哺乳动物宿主中ANP32A之间的差异解释了保护猪和人类免受禽流感病毒人畜共患感染的宿主范围屏障之一。禽流感病毒RNA依赖性RNA聚合酶无法利用哺乳动物ANP 32 A蛋白，除非它通过病毒PB 2蛋白的突变来适应。我们以前已经表明，在飞行的鸟类中存在的蛋白质ANP 32 A的形式不同于在哺乳动物中，由于存在一个额外的外显子编码33个氨基酸，有助于成熟的蛋白质。我们有初步的数据表明，删除这个外显子减少或禽流感病毒在鸡细胞中的复制。我们将确定病毒蛋白相互作用所需的ANP32A的特定区域。我们将使用我们开发的新的基因组编辑工具来修改鸡细胞，以测试我们对ANP32A的靶向变化将对禽流感病毒复制产生最显著影响的预测。我们将产生一系列的ANP32A突变，并测试它们对禽流感聚合酶功能和病毒复制的影响。我们将确定含有修饰的ANP32A蛋白的编辑细胞的RNA转录组中的任何全局变化。我们将研究ANP32A在基因组编辑动物中的功能。这些信息将为保护商业家禽免受AIV感染的控制策略提供信息。研究人类和牲畜流感病毒的研究人员也会感兴趣。

Influenza viruses infect domestic animals including poultry and pigs, causing loss of income, social and economic disruption. Avian influenza remains one of the major threats to food security. Adaptation of avian influenza in animal hosts leads to viruses that are transmitted in humans and carries the risk of driving emergent influenza pandemics. We have investigated a family of host proteins, ANP32, that are co-opted by influenza virus to support its replication. The difference between ANP32A in chickens and mammalian hosts accounts for one of the host range barriers that protect pigs and humans from zoonotic infections by avian influenza viruses. Avian influenza virus RNA-dependent RNA polymerase is unable to utilise the mammalian ANP32A protein unless it adapts by mutation of the viral PB2 protein. We have previously shown that the form of the protein ANP32A present in flighted birds differs from that in mammals due to presence of an additional exon encoding 33 amino acids that contribute to the mature protein. We have preliminary data suggesting that removal of this exon reduces or avian influenza virus replication in chicken cells. We will identify the specific regions of ANP32A needed for viral protein interactions. We will use new genome editing tools that we have developed to modify chicken cells to test our predictions of which targeted changes to ANP32A will have the most significant effect on avian influenza virus replication. We will generate a series of mutations of ANP32A and test their effects on avian influenza polymerase function and virus replication. We will determine any global changes in the RNA transcriptome of the edited cells containing modified ANP32A protein. We will investigate the function of ANP32A in genome edited animals. This information will inform control strategies for protection of commercial poultry from AIV infection. It will also be of interest to researchers studying influenza virus in humans and livestock.