Nck-1 Interacts with PKR and Modulates Its Activation by DsRNA

Activation of the double-stranded RNA (dsRNA)-activated protein kinase PKR results in inhibition of general translation through phosphorylation of the eukaryotic initiation factor 2 alpha-subunit on serine 51 (eIF2alphaSer51). Previously, we have reported that the adaptor protein Nck-1 modulates eIF2alphaSer51 phosphorylation by a subset of eIF2alpha kinases, including PKR. Herein, we demonstrate that Nck-1 prevents efficient activation of PKR by dsRNA, revealing that Nck-1 acts at the level of PKR. In agreement, Nck-1 impairs p38MAPK activation and attenuates cell death induced by dsRNA, in addition to diminish eIF2alphaSer51 phosphorylation. Our data show that the inhibitory effect of Nck-1 on PKR is reversible, as it could be overcome by increasing levels of dsRNA. Interestingly, we found that Nck-1 interacts with the inactive form of PKR, independently of its Src homology domains. Furthermore, we uncovered that Nck-1 is substrate of PKR in vitro. All together, our data provide the first evidence identifying Nck-1 as a novel endogenous regulator of PKR and support the notion that Nck-1-PKR interaction could be a way to limit PKR activation.