Extended Data Table 1.
a, Yeast strains used in this study. b, Cryo-EM data collection, processing and refinement statistics*.
| Strain | Genotype | Source |
|---|---|---|
| TP161 | med7+::TAP-kanMX6 med13::kanMX4 | 18 |
| SG101 | med7+::TAP-kanMX6 med13::kanMX4 med2::clonNAT | This study |
| SG102 | med7+::TAP-kanMX6 med13::kanMX4 med27::clonNAT | This study |
| SG103 | med4+::FLAG-kanMX6 | This study |
| SG104 | med7+::FLAG-kanMX6 | This study |
| SG105 | Med2+::FLAG-kanMX6 | This study |
| SG106 | Med15+::FLAG-kanMX6 | This study |
| KSC101 | BJ2168, Med22-TAP::KlTRP1 | 5 |
| KSC102 | BJ2168, Med22-TAP::KlTRP1 Med31-5xFLAG::KanMX6 | This study |
| KSC103 | BJ2168, Med22-TAP::KlTRP1 Med31(Y38A)-5xFLAG::KanMX6 | This study |
| KSC104 | BJ2168, Med22-TAP::KlTRP1 Med31(Y41E)-5xFLAG::KanMX6 | This study |
| KSC105 | BJ2168, Med22-TAP::KlTRP1 Med31(T44P)-5xFLAG::KanMX6 | This study |
| KSC106 | BJ2168, Med22-TAP::KlTRP1 Med31(Q45A/Q46A)-5xFLAG::KanMX6 | This study |
| KSC107 | BJ2168, Med22-TAP::KlTRP1 Med31(K83A)-5xFLAG::KanMX6 | This study |
| KTB218 | BJ2168, Med22-TAP::KlTRP1 med31Δ::KanMX6 | 5 |
| Data collection and Processing | ||
|---|---|---|
| Microscope | Titan Krios | |
| Voltage (keV) | 300 | |
| Defocus range (μm) | 0.8 to 4.0 | |
| Videos | 11,181 | |
| Frames per movie | 35 | |
| Exposure time per frames (ms) | 200 | |
| Magnification | 22,500x | |
| Dose rate (e−pixel/sec) | 9.8 | |
| Total dose per movie (e−/Å2) | 43 | |
| Refinement | Mediator | Holoenzyme* |
| Particles | 42,484 | 3,862 |
| Map Resolution (Å) | 4.4 | 7.8 |
| RMS angle (°) | 0.01 | 0.01 |
| RMA bonds (Å) | 1.50 | 1.01 |
| Ramachandran outliers (%) | 1.1 | 1.3 |
| Ramachandran allowed (%) | 14.0 | 15.6 |
| Ramachandran favored (%) | 84.9 | 83.1 |
| All-atom clash score | 17.52 | 29.12 |
| Molprobity score | 2.31 | 2.58 |
*
Model statistics for the holoenzyme are based only on the Mediator portion of the model. Resolution for the Mediator portion of the holoenzyme map is 7.4 Å