title: Molecular framework for the activation of RNA-dependent protein kinase.
abstract: The RNA-dependent protein kinase (PKR) plays an integral role in the antiviral response to cellular infection. PKR contains three distinct domains consisting of two conserved N-terminal double-stranded RNA (dsRNA)-binding domains, a C-terminal Ser-Thr kinase domain, and a central 80-residue linker. Despite rich structural and biochemical data, a detailed mechanistic explanation of PKR activation remains unclear. Here we provide a framework for understanding dsRNA-dependent activation of PKR using nuclear magnetic resonance spectroscopy, dynamic light scattering, gel filtration, and autophosphorylation kinetics. In the latent state, PKR exists as an extended monomer, with an increase in self-affinity upon dsRNA association. Subsequent phosphorylation leads to efficient release of dsRNA followed by a greater increase in self-affinity. Activated PKR displays extensive conformational perturbations within the kinase domain. We propose an updated model for PKR activation in which the communication between RNA binding, central linker, and kinase domains is critical in the propagation of the activation signal and for PKR dimerization.
authors.affiliation: Department of Structural Biology and Stanford Magnetic Resonance Laboratory, Stanford University School of Medicine, Stanford, California 94305-5126, USA.
authors.forename: Sean A
authors.initials: SA
authors.lastname: McKenna
authors.forename: Darrin A
authors.initials: DA
authors.lastname: Lindhout
authors.forename: Insil
authors.initials: I
authors.lastname: Kim
authors.forename: Corey W
authors.initials: CW
authors.lastname: Liu
authors.forename: Vladimir M
authors.initials: VM
authors.lastname: Gelev
authors.forename: Gerhard
authors.initials: G
authors.lastname: Wagner
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Puglisi
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D019281:Dimerization; D004789:Enzyme Activation; D006801:Humans; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D010766:Phosphorylation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012330:RNA, Double-Stranded / Q000378:metabolism; D019892:eIF-2 Kinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D012330:RNA, Double-Stranded; D000255:Adenosine Triphosphate; D019892:eIF-2 Kinase