Department of Biology, University of North Carolina, Greensboro, North Carolina 27402, USA.
Genetics. 2011 Oct;189(2):495-506. doi: 10.1534/genetics.111.132290. Epub 2011 Jul 20.
Influenza viruses impose a constant threat to vertebrates susceptible to this family of viruses. We have developed a new tool to study virus-host interactions that play key roles in viral replication and to help identify novel anti-influenza drug targets. Via the UAS/Gal4 system we ectopically expressed the influenza virus M2 gene in Drosophila melanogaster and generated dose-sensitive phenotypes in the eye and wing. We have confirmed that the M2 proton channel is properly targeted to cell membranes in Drosophila tissues and functions as a proton channel by altering intracellular pH. As part of the efficacy for potential anti-influenza drug screens, we have also demonstrated that the anti-influenza drug amantadine, which targets the M2 proton channel, suppressed the UAS-M2 mutant phenotype when fed to larvae. In a candidate gene screen we identified mutations in components of the vacuolar V1V0 ATPase that modify the UAS-M2 phenotype. Importantly, in this study we demonstrate that Drosophila genetic interactions translate directly to physiological requirements of the influenza A virus for these components in mammalian cells. Overexpressing specific V1 subunits altered the replication capacity of influenza virus in cell culture and suggests that drugs targeting the enzyme complex via these subunits may be useful in anti-influenza drug therapies. Moreover, this study adds credence to the idea of using the M2 "flu fly" to identify new and previously unconsidered cellular genes as potential drug targets and to provide insight into basic mechanisms of influenza virus biology.
流感病毒不断威胁着易感染这类病毒的脊椎动物。我们开发了一种新工具,用于研究在病毒复制过程中起关键作用的病毒-宿主相互作用,并帮助鉴定新型抗流感药物靶标。通过 UAS/Gal4 系统,我们在果蝇中异位表达了流感病毒 M2 基因,并在眼睛和翅膀中产生了剂量敏感表型。我们已经证实,M2 质子通道在果蝇组织中被正确靶向细胞膜,并通过改变细胞内 pH 值起质子通道的作用。作为潜在抗流感药物筛选的功效的一部分,我们还证明了抗流感药物金刚烷胺,其靶向 M2 质子通道,当喂食幼虫时抑制 UAS-M2 突变表型。在候选基因筛选中,我们鉴定了空泡 V1V0 ATP 酶组件中的突变,这些突变改变了 UAS-M2 表型。重要的是,在这项研究中,我们证明了果蝇遗传相互作用直接转化为流感 A 病毒在哺乳动物细胞中对这些成分的生理需求。过表达特定的 V1 亚基改变了流感病毒在细胞培养中的复制能力,并表明通过这些亚基针对酶复合物的药物可能在抗流感药物治疗中有用。此外,这项研究为使用 M2“流感果蝇”来鉴定新的和以前未被考虑的细胞基因作为潜在的药物靶标提供了依据,并深入了解流感病毒生物学的基本机制。