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Figure 5.
NEDD1 does not influence the conformation of the γ-TuRC. (A) Two cartoon representation views of superimposed rec-γ-TuRC (grey) and rec-γ-TuRC + CDK5RAP2 (magenta) models determined in this study. The right panel shows a close-up view of the NEDD1 pinwheel and its binding interface with the GRIP1 domains of GCP4–6 at positions 9–12. (B) Two cartoon representation views of superimposed rec-γ-TuRC (grey) and a “partially-closed”, CMG-decorated γ-TuRC (blue; PDB: 9G3Y [Xu et al., 2024]). The right panel shows a close-up view of the NEDD1 pinwheel and its location relative to the GRIP1 domains of GCPs modeled at positions 9–12. (C) Two cartoon representation views of superimposed rec-γ-TuRC (grey) and the “fully-closed” rec-γ-TuRC (green; PDB: 8VRK [Aher et al., 2024]). The right panel shows a close-up view of the NEDD1 pinwheel and its location relative to the GRIP1 domains of GCPs modeled at positions 9–12. RMSD values for γ-tubulins at position 14 (left) and position 9–12 GCP GRIP1 domains (right) are indicated in panels A–C. Asterisk in panel C is to clarify that the GRIP1 domains in 8VRK correspond not to GCP4/5/6 but to GCP2/3 models, built into an 8.5-Å reconstruction (Aher et al., 2024), both of which might potentially limit the accuracy of the RMSD measurement in this example. (D) Plot of Euclidean center of mass distances (dCOM) versus γ-TuRC subunit position between the indicated γ-TuRC models (rec-γ-TuRC, rec-γ-TuRC + CDK5RAP2, PDB: 6V6S as the open conformation (Wieczorek et al., 2020b), and the “partially-closed,” CMG-decorated (Xu et al., 2024), all relative to PDB: 8VRK, corresponding to a model of the closed rec-γ-TuRC (Aher et al., 2024). (E) Plot of the average shift in θ versus the shift in ϕ for helix H12 in γ-tubulins from each γ-TuRC described in D, relative to γ-tubulins at the same positions in the closed γ-TuRC (green circle, indicated). Standard errors in ϕ and θ are displayed as lines. The axes in E are scaled equally. Coloring in E follows the legend in D. (F) Model summarizing the findings in this study. The rec-γ-TuRC + CDK5RAP2 model has been converted to a 15-Å low-pass filtered map and colored according to Fig. 4 B. Unresolved WD40 domains stemming from the NEDD1 pinwheel and available to interact with centrosomes, microtubules, augmin, and/or other partners are shown as hexagons. Free CMG module–binding sites that should still be able to induce γ-tubulin ring closure in the presence of the NEDD1 pinwheel are indicated. RSMD, root mean squared deviation.

NEDD1 does not influence the conformation of the γ-TuRC. (A) Two cartoon representation views of superimposed rec-γ-TuRC (grey) and rec-γ-TuRC + CDK5RAP2 (magenta) models determined in this study. The right panel shows a close-up view of the NEDD1 pinwheel and its binding interface with the GRIP1 domains of GCP4–6 at positions 9–12. (B) Two cartoon representation views of superimposed rec-γ-TuRC (grey) and a “partially-closed”, CMG-decorated γ-TuRC (blue; PDB: 9G3Y [Xu et al., 2024]). The right panel shows a close-up view of the NEDD1 pinwheel and its location relative to the GRIP1 domains of GCPs modeled at positions 9–12. (C) Two cartoon representation views of superimposed rec-γ-TuRC (grey) and the “fully-closed” rec-γ-TuRC (green; PDB: 8VRK [Aher et al., 2024]). The right panel shows a close-up view of the NEDD1 pinwheel and its location relative to the GRIP1 domains of GCPs modeled at positions 9–12. RMSD values for γ-tubulins at position 14 (left) and position 9–12 GCP GRIP1 domains (right) are indicated in panels A–C. Asterisk in panel C is to clarify that the GRIP1 domains in 8VRK correspond not to GCP4/5/6 but to GCP2/3 models, built into an 8.5-Å reconstruction (Aher et al., 2024), both of which might potentially limit the accuracy of the RMSD measurement in this example. (D) Plot of Euclidean center of mass distances (dCOM) versus γ-TuRC subunit position between the indicated γ-TuRC models (rec-γ-TuRC, rec-γ-TuRC + CDK5RAP2, PDB: 6V6S as the open conformation (Wieczorek et al., 2020b), and the “partially-closed,” CMG-decorated (Xu et al., 2024), all relative to PDB: 8VRK, corresponding to a model of the closed rec-γ-TuRC (Aher et al., 2024). (E) Plot of the average shift in θ versus the shift in ϕ for helix H12 in γ-tubulins from each γ-TuRC described in D, relative to γ-tubulins at the same positions in the closed γ-TuRC (green circle, indicated). Standard errors in ϕ and θ are displayed as lines. The axes in E are scaled equally. Coloring in E follows the legend in D. (F) Model summarizing the findings in this study. The rec-γ-TuRC + CDK5RAP2 model has been converted to a 15-Å low-pass filtered map and colored according to Fig. 4 B. Unresolved WD40 domains stemming from the NEDD1 pinwheel and available to interact with centrosomes, microtubules, augmin, and/or other partners are shown as hexagons. Free CMG module–binding sites that should still be able to induce γ-tubulin ring closure in the presence of the NEDD1 pinwheel are indicated. RSMD, root mean squared deviation.

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