Influenza virus assembly and virulence

超出提供的空间。甲型流感病毒的组装涉及病毒感染细胞内预定位置的几种病毒成分的特定相互作用。参与这一过程的病毒蛋白是疫苗生成以及病毒适应新宿主的关键成分，但组装过程中的确切作用尚未确定或确定。反向遗传学技术和新技术(如RNA干扰)的应用使人们能够精确地剖析组装过程，并确定对病毒组装重要的精确功能域。为了充分了解甲型流感病毒组装所涉及的病毒因素，将对参与病毒组装的M1(基质)和M2蛋白的结构域进行详细的分子研究。将识别M1和M2中的氨基酸序列，这些氨基酸序列在将蛋白质定位到病毒萌发的位置方面起到重要作用。抗M2抗体介导的抑制甲型流感病毒萌发的机制将被阐明，M1蛋白和M2细胞质尾部之间的特定相互作用将被表征。最后，将在感染了携带HA和NA蛋白突变的重组病毒的细胞中分析M1的细胞内定位，这些突变阻止了它们与糖脂筏或顶膜的联系。将产生含有一种或几种病毒蛋白特定突变的重组病毒，并对其对抗M2抗体的敏感性、抗病毒药物敏感性和毒力进行表征。这些研究应该会进一步阐明导致新的高毒力甲型流感病毒产生的机制，并进一步确定支配抗体和药物敏感性的相互作用。表演网站========================================Section End===========================================

EXCEED THE SPACE PROVIDED. The assembly of influenza A virus involves the specific interaction of several viral components at a predetermined location within the virus-infected cell. The viral proteins involved in this process are key components in the generation of vaccines as well as in the adaption of virus to new hosts but precise roles in the assembly process have yet to be ascribed or identified. The application of reverse genetics techniques and new technologies such as RNA interference allows for the precise dissection of the assembly process and the determination of precise functional domains important for virus assembly to take place. To fully understand the viral factors involved in influenza A virus assembly, a detailed molecular investigation of the domains of the M1 (matrix) and M2 proteins involved in virus assembly will be undertaken. The amino acid sequences in M1 and M2 that are important in targeting the proteins to sites of virus budding will be identified. The mechanism behind the anti-M2 antibody mediated inhibition of influenza A virus budding will be elucidated and specific interactions between the M1 protein and the M2 cytoplasmic tail will be characterized. Finally, the intracellular localization of M1 will be analyzed in cells infected with recombinant viruses bearing mutations in the HA and NA proteins that prevent their association with glycolipid rafts or the apical membrane. Recombinant viruses bearing specific mutations in one or several viral proteins will be generated and characterized for their sensitivity to anti-M2 antibodies, antiviral drug sensitivity and virulence. These studies should shed additional light on the mechanisms responsible for the generation of new, highly virulent influenza A viruses as well as further define the interactions that govern antibody and drug sensitivity. PERFORMANCE SITE ========================================Section End===========================================