The Research Of Avian Influenza (H5N1) Of The NS1Protein Interaction Sites With The Host Protein NOLC1

Avian influenza that is caused by Avian influenza virus(AIV)is a highlycontagious acute infectious disease from respiratory system pathological changes tosepticaemic of total body, Avian influenza have a direct threat to human health sinceit can spread from poultry to humans, there is seriously affecting the survival anddevelopment of the aquaculture industry, and cause considerable harm to humanpublic health. NS1protein (nonstructural protein1) is one of the importantnon-structural protein of type A influenza virus, it is exists in the nucleus only in thevirus infected cells in the early stages. A-type influenza virus NS1protein expressedat high levels in infected cells, its main function is to inhibit the IFN-a/b(IFN)-mediated antiviral activity, promote the expression of viral genes andinhibition of host mRNA processing.Human nucleolar and coiled-body phosphoprotein1(NOLC1), is a highlyphosphorylated protein, it was verified interacted with NS1, and it plays an importantrole in the process of the occurrence of the nucleolus, cell division and apoptosis.Our preliminary work has got a host protein NOLC1which could interacted withNS1protein by using co-immunoprecipitation and His-pull down in vitro and in vivoto verify the interaction between the two proteins.Fluorescence positioningtechnology was used to the two co-localized in the nucleus. On this basis, the genetruncation techniques was used to determine the effect of the NS1protein domain (i.e.NS1-C) with protein-NOLC1interaction. The study is based on the basis of previousexperiments, we use the genetic truncation, PCR directed mutagenesis and His-pulldown technology continue to study NS1-C region, and directed mutagenesis inNS1-C.The spatial structure of NS1-C terminal area were analyzed, and truncated bygene technology, to further determine the NS1-C terminal area and accurate area ofNOLC1protein interaction is NS1-C area104-200amino acid protein area; usingPCR site-directed mutation technology in vitro to single point mutations of NS1-C,determine the NS1-C120-D and195-R acid are key sites involved in NS1-C interactwith NOLC1. This study to explore designed for the study of antiviral drugs targeted to provide important reference and also for further study the structure and thefunction of NOLC1provides the help.