title: The activation of the decapping enzyme DCP2 by DCP1 occurs on the EDC4 scaffold and involves a conserved loop in DCP1.
abstract: The removal of the 5'-cap structure by the decapping enzyme DCP2 and its coactivator DCP1 shuts down translation and exposes the mRNA to 5'-to-3' exonucleolytic degradation by XRN1. Although yeast DCP1 and DCP2 directly interact, an additional factor, EDC4, promotes DCP1-DCP2 association in metazoan. Here, we elucidate how the human proteins interact to assemble an active decapping complex and how decapped mRNAs are handed over to XRN1. We show that EDC4 serves as a scaffold for complex assembly, providing binding sites for DCP1, DCP2 and XRN1. DCP2 and XRN1 bind simultaneously to the EDC4 C-terminal domain through short linear motifs (SLiMs). Additionally, DCP1 and DCP2 form direct but weak interactions that are facilitated by EDC4. Mutational and functional studies indicate that the docking of DCP1 and DCP2 on the EDC4 scaffold is a critical step for mRNA decapping in vivo. They also revealed a crucial role for a conserved asparagine-arginine containing loop (the NR-loop) in the DCP1 EVH1 domain in DCP2 activation. Our data indicate that DCP2 activation by DCP1 occurs preferentially on the EDC4 scaffold, which may serve to couple DCP2 activation by DCP1 with 5'-to-3' mRNA degradation by XRN1 in human cells.
authors: Department of Biochemistry, Max Planck Institute for Developmental Biology, Spemannstrasse 35, 72076 Tübingen, Germany.
authors: Chung-Te
authors: CT
authors: Chang
authors: Natalia
authors: N
authors: Bercovich
authors: Belinda
authors: B
authors: Loh
authors: Stefanie
authors: S
authors: Jonas
authors: Elisa
authors: E
authors: Izaurralde
mesh_terms: D000595:Amino Acid Sequence; D017124:Conserved Sequence; D004722:Endoribonucleases / Q000737:chemistry* / Q000378:metabolism*; D005095:Exoribonucleases / Q000378:metabolism; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D010649:Phenylalanine / Q000032:analysis; D054730:Protein Interaction Domains and Motifs; D011506:Proteins / Q000378:metabolism*; D015534:Trans-Activators / Q000737:chemistry* / Q000378:metabolism*
chemical_list: C507914:EDC4 protein, human; D008869:Microtubule-Associated Proteins; D011506:Proteins; D015534:Trans-Activators; D010649:Phenylalanine; D004722:Endoribonucleases; D005095:Exoribonucleases; C470512:XRN1 protein, human; C466279:DCP1A protein, human; C467723:DCP2 protein, human