Figure 5. CryoET of UL25-mediated inhibition of NEC budding.
(a) CryoET averages of NEC220 in the presence of UL25Δ44 Q72A (top and side views). Corresponding 3D models are shown with NEC220 (pink) and UL25Δ44 Q72A (green). The vesicle bilayer is shown in beige. The models show the UL25 layer coating the NEC layer in five-pointed stars on the outside of the vesicles. The length of the NEC-UL25 spikes is 12.5 nm. Black arrows indicate the point of tilt within the NEC layer. (b) CryoET averages of NEC220 forming hexameric lattices in the presence of membranes (bottom and side views). Corresponding 3D models are shown with NEC (pink) and the vesicle bilayer (beige). The diameter of the hexameric rings is ~11 nm, while the length of the spikes is 7.8 nm. (c) CryoET model and averages of the UL25 layer (green) highlighting the five-pointed star formation of UL25 (represented here as a pentamer of dimers) in the presence of NEC. (d) CryoET model and averages of the NEC layer showing NEC220 forming a pentagonal lattice (pink pentagons), rather than hexagonal (as seen for wild-type in panel b). Green triangles indicate location of UL25 binding to the NEC.