tailieunhanh - Báo cáo y học: "Mutagenesis analysis of the zinc-finger antiviral protein"

Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học Retrovirology cung cấp cho các bạn kiến thức về ngành y đề tài: "Mutagenesis analysis of the zinc-finger antiviral protein. | Wang et al. Retrovirology 2010 7 19 http content 7 1 19 gtr RETR0VIR0L0GY RESEARCH Open Access Mutagenesis analysis of the zine-finger antiviral protein Xinlu Wang Fengxiang Lv and Guangxia Gao Abstract Background The zinc-finger antiviral protein ZAP specifically inhibits the replication of certain viruses including murine leukemia virus MLV by preventing the accumulation of viral mRNA in the cytoplasm. ZAP directly binds to the viral mRNA through the zinc-finger motifs and recruits the RNA exosome to degrade the target RNA. RNA helicase p72 is required for the optimal function of ZAP. In an attempt to understand the structure-function relationship of ZAP we performed alanine scanning analysis. Results A series of ZAP mutants was generated in which three consecutive amino acids were replaced with three alanines. The mutants were analyzed for their antiviral activities against pseudotyped MLV vector. Out of the nineteen mutants analyzed seven displayed significantly lower antiviral activities. Two mutations were in the very N-terminal domain and five mutations were within or around the first and second zinc-finger motifs. These mutants were further analyzed for their abilities to bind to the target RNA the exosome and the RNA helicase p72. Mutants Nm3 and Nm63 lost the ability to bind to RNA. Mutants Nm 63 and Nm93 displayed compromised interaction with p72 while the binding of Nm133 to p72 was very modest. The interactions of all the mutants with the exosome were comparable to wild type ZAP. Conclusions The integrity of the very N-terminal domain and the first and second zinc-finger motifs appear to be required for ZAP s antiviral activity. Analyses of the mutants for their abilities to interact with the target RNA and RNA helicase p72 confirmed our previous results. The mutants that bind normally to the target RNA the exosome and the RNA helicase p72 may be useful tools for further understanding the mechanism underlying ZAP s antiviral .