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Figure 3.
The NEDD1 pinwheel contacts an extension in the GCP6 belt and enables the assignment of MZT1:GCP5-NHD to the γ-TuRC seam. (A) Two views of the upper helical pair of the NEDD1 fishtail contacting positions 1 and 2 of the γ-TuRC (top: cartoon representation and cryo-EM density in transparent grey surface; middle: cartoon representation of NEDD1 and GCP6 next to a surface representation of the γ-TuRC). Newly modeled GCP6 residues 191–252 extending from the GCP6 belt are indicated. The bottom panel shows a rotated view of the same interface to highlight the unattached helical pair in the fishtail. γ-TuRC subunit positions 1 (GCP2) and 2 (GCP3) are indicated, where possible. (B) AlphaFold model of GCP3, MZT1:GCP5-NHD, and γ-tubulin rigid-body fitted in the rec-γ-TuRC consensus map (transparent representation; right). The model on the left is colored according to the pLDDT score from the AlphaFold prediction (cartoon representation). The model in the middle is shown in surface representation and is colored according to the legend. (C) AlphaFold model of GCP3, MZT1:GCP3-NHD, and γ-tubulin rigid-body fitted in the rec-γ-TuRC consensus map (transparent representation; right). The model on the left is colored according to the pLDDT score from the AlphaFold prediction (cartoon representation). The model in the middle is shown in surface representation and is colored according to the legend. (D) Rigid body-fitted AlphaFold model of GCP5, GCP4, GCP6, GCP2, GCP3, and MZT1 in the rec-γ-TuRC consensus map (transparent representation). MZT1:GCP5-NHD, GCP3, and the disordered GCP5 linker (aa 120–200) are indicated. The Euclidean distance between GCP5 residues 120 and 200 in the model is indicated. (E) Secondary structure prediction of human GCP5 residues 120–200. Top: primary sequence (red = predicted α-helices); middle: jnetpred secondary structure prediction result (red = α-helices); bottom: confidence score for the prediction. Figure panel generated using Jalview (Waterhouse et al., 2009). pLDDT, predicted local distance difference test.

The NEDD1 pinwheel contacts an extension in the GCP6 belt and enables the assignment of MZT1:GCP5-NHD to the γ-TuRC seam. (A) Two views of the upper helical pair of the NEDD1 fishtail contacting positions 1 and 2 of the γ-TuRC (top: cartoon representation and cryo-EM density in transparent grey surface; middle: cartoon representation of NEDD1 and GCP6 next to a surface representation of the γ-TuRC). Newly modeled GCP6 residues 191–252 extending from the GCP6 belt are indicated. The bottom panel shows a rotated view of the same interface to highlight the unattached helical pair in the fishtail. γ-TuRC subunit positions 1 (GCP2) and 2 (GCP3) are indicated, where possible. (B) AlphaFold model of GCP3, MZT1:GCP5-NHD, and γ-tubulin rigid-body fitted in the rec-γ-TuRC consensus map (transparent representation; right). The model on the left is colored according to the pLDDT score from the AlphaFold prediction (cartoon representation). The model in the middle is shown in surface representation and is colored according to the legend. (C) AlphaFold model of GCP3, MZT1:GCP3-NHD, and γ-tubulin rigid-body fitted in the rec-γ-TuRC consensus map (transparent representation; right). The model on the left is colored according to the pLDDT score from the AlphaFold prediction (cartoon representation). The model in the middle is shown in surface representation and is colored according to the legend. (D) Rigid body-fitted AlphaFold model of GCP5, GCP4, GCP6, GCP2, GCP3, and MZT1 in the rec-γ-TuRC consensus map (transparent representation). MZT1:GCP5-NHD, GCP3, and the disordered GCP5 linker (aa 120–200) are indicated. The Euclidean distance between GCP5 residues 120 and 200 in the model is indicated. (E) Secondary structure prediction of human GCP5 residues 120–200. Top: primary sequence (red = predicted α-helices); middle: jnetpred secondary structure prediction result (red = α-helices); bottom: confidence score for the prediction. Figure panel generated using Jalview (Waterhouse et al., 2009). pLDDT, predicted local distance difference test.

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