Structure of the Prefusion Form of the Vesicular Stomatitis Virus Glycoprotein G
Stéphane Roche, Félix A. Rey, Yves Gaudin, Stéphane Bressanelli

Supporting Online Material

This supplement contains:
Materials and Methods
Figs. S1 to S5
Table S1
References
Movie S1

This file is in Adobe Acrobat PDF format.

Movie 1
A transition from the pre- to post-fusion forms was generated by keeping the invariant RbI-II (table I and figure 1B) fixed and relocating DIII, DIV and the C-terminus around it, as in the text (Fig. 1B). For clarity, the movie was made in 3 successive stages: swinging of DIV around the DIII/DIV connection first; then refolding of the DII/DIII connection; finally, refolding of the C-terminus. While analysis of the structures suggests that this order is correct in outline (see below), the stages may partially overlap in the actual change.

Stage 1 was morphed in one step, while stages 2 and 3 were broken up into 3 steps each. For each step, n substeps (n=40-100) of interpolation/minimization were performed using CNS (S13) and scripts adapted from the script described in the Yale Molecular Morphing Server web site [(S14), http://www.molmovdb.org/molmovdb/morph/]. The intermediates were used to generate a movie with PyMOL (S15).

Stage 1 has to occur first, as the interactions depicted in figure 4A have to be broken before stages 2 and 3 can start.

Stage 2: Once DIV has moved out of the way, helix F2 can start refolding into helix F (figure 2B). Step 1 is the lengthening of the N-terminus of helix F1. Step 2 is a rotation around the connection to F2. These two steps free the segment downstream of strand a1, that is sandwiched in the prefusion conformation between F1 and RbI-II, to refold into helix A. This and the completion of helix F constitute step 3.

Stage 3: Breaking up of the histidine cluster shown in figure 4A, bottom, frees the segment directly upstream, including y2 and H2 (figure 2B), to move out of the bottom crevice in RbIII (figure 2C). Step 1 is a ca 60° rotation around the H1-y1 connection. This movement breaks up the q1y2 sheet. Step 2 completes the refolding of the H1-y1-H2 segment into helix H. In step 3, helix H rotates 90° relative to RbI-II to pack against helix F. This large movement is achieved by rotating the single conserved P383 (figure S2) around the F382 and P383 psi angles.

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