López-Perrote…Llorca {Nucleic Acids Res 44: 1909}
3D model of the complex between nonsense-mediated mRNA decay factor UPF2 and eukaryotic release factor eRF3. The model was obtained using electron microscopy (EM) of purified eRF3-UPF2 complexes, and atomic structures of some domains were fitted into the EM model. One of the major findings in Lopez-Perrote et al. is the discovery that UPF2 interacts with eRF3 and the biochemical and structural characterisation of this complex.
Human nonsense-mediated mRNA decay factor UPF2 interacts directly with eRF3 and the SURF complex
López-Perrote A, Castaño R, Melero R, Zamarro T, Kurosawa H, Ohnishi T, Uchiyama A, Aoyagi K, Buchwald G, Kataoka N, Yamashita A, Llorca O.
Nucleic Acids Res 2016 Feb; 44: 1909.
Nonsense-mediated mRNA decay (NMD) is an mRNA degradation pathway that regulates gene expression and mRNA quality. A complex network of macromolecular interactions regulates NMD initiation, which is only partially understood. According to prevailing models, NMD begins by the assembly of the SURF (SMG1-UPF1-eRF1-eRF3) complex at the ribosome, followed by UPF1 activation by additional factors such as UPF2 and UPF3. Elucidating the interactions between NMD factors is essential to comprehend NMD, and here we demonstrate biochemically and structurally the interaction between human UPF2 and eukaryotic release factor 3 (eRF3). In addition, we find that UPF2 associates with SURF and ribosomes in cells, in an UPF3-independent manner. Binding assays using a collection of UPF2 truncated variants reveal that eRF3 binds to the C-terminal part of UPF2. This region of UPF2 is partially coincident with the UPF3-binding site as revealed by electron microscopy of the UPF2-eRF3 complex. Accordingly, we find that the interaction of UPF2 with UPF3b interferes with the assembly of the UPF2-eRF3 complex, and that UPF2 binds UPF3b more strongly than eRF3. Together, our results highlight the role of UPF2 as a platform for the transient interactions of several NMD factors, including several components of SURF.
PubMed: 26740584. Doi: 10.1093/nar/gkv1527
