PKR in innate immunity, cancer, and viral oncolysis.

摘要: The mammalian innate immune system provides a first line of defense against microbial pathogens and also serves to activate an antigen specific acquired program. Key components immunity are the interferons (IFNs), family related cytokines with potent antimicrobial immuno-modulatory activities. IFNs exert their effects through induction numerous genes, one which is double-stranded RNA-dependent protein kinase (PKR), pivotal antiviral found in most human cells. Following activation by double stranded (ds) RNAs produced during viral replication, PKR phosphorylates alpha-subunit eukaryotic translation initiation factor (eIF) 2, causing severe inhibititon cellular synthesis. Phosphorylation eIF2alpha consequent inhibition synthesis major cell growth checkpoint utilized at least three other kinases, addition PKR, following exposure such stresses as amino acid deprivation presence misfolded proteins endoplasmic reticulum. Indeed, it has been demonstrated that disruption can lead transformation immortalized rodent cells, plausibly increasing rates proto-oncogenes. Further, shown disregulation permissiveness virus infection may be common occurrence tumorigenic lines. These findings have exploited develop oncolytic RNA viruses selectively replicate destroy variety neoplasias vitro vivo. In this chapter, we describe some techniques commonly used our laboratory examine activity eIF2 regulation. Protocols for generation use recombinant vesicular stomatitis variants described.