Table 1.
Effect of polymerization conditions on Sp_tub MT architecture
| S. pombe tubulin polymerization conditionsa | PF number | moiré repeat (nm) (± ~2 nm) | MT number (%) | |
|---|---|---|---|---|
| GTP | −Mal3 (n = 25) | 12 | 140–224 | 2 (8%) |
| “ | 13 | 210– > 1000 | 21 (84%) | |
| “ | 14 | 505–624 | 2 (8%) | |
| + Mal3 (n = 10) | 13 | 400– > 1000 | 8 (80%) | |
| “ | 14 | > 1000 | 1 (10%) | |
| “ | 15 | > 1000 | 1 (10%) | |
| GMPCPP | −Mal3 (n = 30) | 11 | 100 | 1 (3%) |
| “ | 12 | 124–156 | 12 (40%) | |
| “ | 13 | 90–144 | 15 (50%) | |
| “ | 14 | 90–100 | 2 (7%) | |
| + Mal3 (n = 19) | 12 | >1000 | 2 (11%) | |
| “ | 13 | 400– > 1000 | 8 (42%) | |
| “ | 14 | 140– > 1000 | 7 (37%) | |
| “ | 15 | 260 | 2 (11%) | |
MT architecture parameters were determined by analysis of each cryo-ET 3D volume, and were found to be consistent with the lattice accommodation model proposed by39,40
aMTs were polymerized as described in the Methods section and KMD was subsequently added prior to cryo-EM sample preparation, apart from the dynamic + GTP MTs, where no KMD was added