The role of the influenza NS1A protein during influenza A virus infection evasion of the host anti-viral response /

by Min, Ji-Young

73 Introduction 74 Materials and Methods 77 Cell lines 77 Antibodies and other reagents 77 Expression vectors 77 Purification of NS1A, PACT and their derivatives 78 PKR kinase assay in vitro 79 Assay for binding of PACT domain 3 to PKR 79 GST-pull down assay 80 Analysis of the phosphorylation of PKR and eIF-2? in vivo 81 Cell survival assay 81 Results 82 The NS1A protein inhibits PKR activation mediated by PACT 82 The binding of the NS1A protein to PKR residues 1-230 is responsible for the inhibition of PACT-mediated activation of PKR 84 The binding of the NS1A protein to PKR blocks its activation by dsRNA as well as by PACT and does not require dsRNA binding by the NS1A protein 88 The NS1A protein inhibits PKR activation by PACT in vivo 88 x Discussion 92