Molecular Basis for Recognition of Dilysine Trafficking Motifs by COPI

Summary

COPI mediates retrograde trafficking from the Golgi to the endoplasmic reticulum (ER) and within the Golgi stack, sorting transmembrane proteins bearing C-terminal KKxx or KxKxx motifs. The structure of KxKxx motifs bound to the N-terminal WD-repeat domain of β'-COP identifies electrostatic contacts between the motif and complementary patches at the center of the β'-COP propeller. An absolute requirement of a two-residue spacing between the terminal carboxylate group and first lysine residue results from interactions of carbonyl groups in the motif backbone with basic side chains of β'-COP. Similar interactions are proposed to mediate binding of KKxx motifs by the homologous α-COP domain. Mutation of key interacting residues in either domain or in their cognate motifs abolishes in vitro binding and results in mistrafficking of dilysine-containing cargo in yeast without compromising cell viability. Flexibility between β'-COP WD-repeat domains and the location of cargo binding have implications for COPI coat assembly.

Highlights

► Dilysine motifs bind the top surface of β'- and α-COP N-terminal WD-repeat domains ► Mutation of key binding site residues abolishes in vitro binding to dilysine motifs ► Loss of binding site prevents retrograde trafficking of dilysine motifs in vivo ► Mutants lacking dilysine motif binding site can transport other retrograde cargoes

Abstract

COPI facilitates trafficking of transmembrane proteins bearing C-terminal KKxx or KxKxx motifs between the Golgi and the ER and within the Golgi stacks. Jackson et al. now provide molecular characterization of COPI cargo binding through elucidation of the structure of β'-COP N-terminal WD-repeat domain in complex with a KxKxx motif.