@article {Travis2020.04.07.029496, author = {Sophie M. Travis and Kevin DAmico and I-Mei Yu and Safraz Hamid and Gabriel Ramirez-Arellano and Philip D. Jeffrey and Frederick M. Hughson}, title = {Structural basis for the binding of SNAREs to the multisubunit tethering complex Dsl1}, elocation-id = {2020.04.07.029496}, year = {2020}, doi = {10.1101/2020.04.07.029496}, publisher = {Cold Spring Harbor Laboratory}, abstract = {Multisubunit tethering complexes (MTCs) are large (250 to \>750 kDa), conserved macromolecular machines that are essential for SNARE-mediated membrane fusion in all eukaryotes. MTCs are thought to function as organizers of membrane trafficking, mediating the initial, long-range interaction between a vesicle and its target membrane and promoting the formation of membrane-bridging SNARE complexes. Previously, we reported the structure of the Dsl1 complex, the simplest known MTC, which is essential for COPI-mediated transport from the Golgi to the endoplasmic reticulum (ER). This structure suggested how the Dsl1 complex might function to tether a vesicle to its target membrane by binding at one end to the COPI coat and at the other end to ER SNAREs. Here, we use x-ray crystallography to investigate these Dsl1-SNARE interactions in greater detail. The Dsl1 complex comprises three subunits that together form a two-legged structure with a central hinge. Our results show that distal regions of each leg bind N-terminal Habc domains of the ER SNAREs Sec20 (a Qb-SNARE) and Use1 (a Qc-SNARE). The observed binding modes appear to anchor the Dsl1 complex to the ER target membrane while simultaneously ensuring that both SNAREs are in open conformations with their SNARE motifs available for assembly. The proximity of the two SNARE motifs, and therefore their ability to enter the same SNARE complex, depends on the relative orientation of the two Dsl1 legs.}, URL = {https://www.biorxiv.org/content/early/2020/04/07/2020.04.07.029496}, eprint = {https://www.biorxiv.org/content/early/2020/04/07/2020.04.07.029496.full.pdf}, journal = {bioRxiv} }