CATCHR, HOPS and CORVET tethering complexes share a similar architecture

Abstract

We show here that the Saccharomyces cerevisiae GARP complex and the Cog1-4 subcomplex of the COG complex, both members of the complexes associated with tethering containing helical rods (CATCHR) family of multisubunit tethering complexes, share the same subunit organization. We also show that HOPS, a tethering complex acting in the endolysosomal pathway, shares a similar architecture, thus suggesting that multisubunit tethering complexes use related structural frameworks.

Figure 1

Subunit organization of the yeast GARP complex and comparison with the Cog1-4 subcomplex. (a) Selected averages of the GARP complex. The three legs of the Y-shaped complex are labeled from the shortest leg (A) to the longest leg (C). The arrowheads point to a hook at the end of leg B and the arrows point to a hinge in the middle of leg C. (b) Selected class averages of GARP complex tagged with GFP at the C terminus of Vps51 (see Supplementary Fig. 1f for all class averages). The GFP (indicated by arrows) is located close to the hinge region in leg C. (c) Raw particle image (top panel) and schematic drawing (bottom panel) of GARP complex with an N-terminal GFP on Vps51. A C-terminal GFP on Vps54 was used to mark leg C. See Supplementary Fig. 3g for additional particles. (d - i) Raw particle images (top panels) and schematic drawings (bottom panels) of GARP complexes with a GFP at the N terminus (d) or C terminus (e) of Vps52, at the N terminus (f) or C terminus (g) of Vps53, and at the N terminus (h) or C terminus (i) of Vps54. A C-terminal GFP on Vps51 was used to mark leg C. See Supplementary Fig. 3a-f for additional particles. Scale bars in panels (a) to (i) are 15 nm. (j, k) Schematic drawings of the GARP complex (j) and the Cog1-4 subcomplex (k), revealing which subunits in the two complexes correspond to each other.

Figure 2

Proposed subunit organization of the HOPS complex and comparison with the GARP complex. (a) Selected averages of the full HOPS complex and different subcomplexes. Averages 1 to 4 illustrate the structural variability of the HOPS complex. In average 1 the structural features of the HOPS complex are marked. The arrow in average 2 points to leg B that usually appears as a smeared-out density. Averages 5 and 6 show subcomplexes missing domain II. Averages 7 and 8 show subcomplexes missing leg A. Averages 9 and 10 show subcomplexes missing leg C. The arrows in panels 5 to 10 point to the area where the missing domains would be located in the full complex. The scale bar is 20 nm. (b, c) Schematic drawings of the HOPS complex (b) and the GARP complex (c), revealing which subunits in the two complexes potentially correspond to each other. Arrows and protein names indicate the protein substitutions that convert the HOPS to the CORVET complex and the GARP to the EARP complex.