Figure 1. The CM1 region forms a conserved coiled-coil hairpin structure. (A) An overlay of AlphaFold models of Cnn aa100–208 (magenta) and CDK5RAP2 aa54–208 (green), where the SCC and LCC subdomains form a near-identical hairpin structure. Key More about this image found in The CM1 region forms a conserved coiled-coil hairpin structure. (A) An ove...

Figure 2. The isolated CAI forms a partially flexible 4-helix dimer. (A) ¹H-¹⁵N SOFAST-HMQC spectrum of the uniformly ¹⁵N–labeled CAI domain (aa1–77). The assigned signals are labeled. (B) CSI profiles for Cα, Cβ, CO, and Hα nuclei of the CAI, More about this image found in The isolated CAI forms a partially flexible 4-helix dimer. (A) ¹...

Figure 3. The CAI can associate with the SCC. (A) Dimensionless Kratky plots of the SAXS experiments for the CAI (aa1–77) (blue, same data as in Fig. S3 C ) and CAI-L-SCC (1–134) (dark blue) after deconvolution. The expected maximum position More about this image found in The CAI can associate with the SCC. (A) Dimensionless Kratky plots of the ...

Figure 4. The CAI associates with the SCC rather than the LCC in the longer construct CAI-L-SCC-LCC. (A and B) AlphaFold models of CAI-L-SCC-LCC (aa1–208) colored using the pLDDT score. The conserved phenylalanine residues are indicated, along More about this image found in The CAI associates with the SCC rather than the LCC in the longer construct...

Figure 5. HDX-MS data support the CAI interacting with the SCC. (A) Deuterium uptake plot for the CAI-L-SCC-LCC construct (aa1–208) showing the relative fractional uptake for peptides spanning residues −5 to 208 (residues −5 to 0 are part of a More about this image found in HDX-MS data support the CAI interacting with the SCC. (A) Deuterium uptak...

Figure 6. The Cnn’s SCC–LCC is required for robust γ-TuRC binding. (A–D) Western blots showing the results of IPs in which various MBP-tagged Cnn or CDK5RAP2 constructs (as indicated above each lane) attached to anti-MBP magnetic beads were More about this image found in The Cnn’s SCC–LCC is required for robust γ-TuRC binding. (A–D) Western blo...

Figure 1. Vid27 is essential in S. japonicus and functions in parallel with Cmp7 to promote NE assembly. (A) vid27-G67R suppresses the growth defect of cmp7Δ cells. Normalized colony size after ascus dissection. Means ± 95% CI are shown. n = More about this image found in Vid27 is essential in S. japonicus and functions in parall...

Figure 2. Loss of Vid27 leads to defects in NE assembly. (A) Vid27 depletion leads to delays or a block to postmitotic NE assembly. Tracks of time-lapse series showing the state of NE integrity from mitotic NE breakdown (time 0) until the end of More about this image found in Loss of Vid27 leads to defects in NE assembly. (A) Vid27 depletion leads t...

Figure 3. Vid27 localizes to the major sites of NE sealing and is required for proper postmitotic NE assembly. (A–C) Depletion of Vid27 results in large NE holes. (A) Representative EM images of Vid27-depleted (+ASV+IAA) and control (+DMSO) More about this image found in Vid27 localizes to the major sites of NE sealing and is required for proper...

Figure 4. The essential function of Vid27 is contained within its N terminus, containing its two PH-like domains. (A) AlphaFold2 model and cartoon of the Vid27 structure. The G67R suppressor mutation is shown in red. (B) Design of Vid27 More about this image found in The essential function of Vid27 is contained within its N terminus, contain...

Figure 5. Vid27 and Alx1 interact and promote NE assembly, independent of ESCRT-III. (A and B) IP MS indicates that Vid27 and Vid27-G67R interact with Alx1. Volcano plots show TMT-based quantitative MS of Vid27-intFLAG and Vid27-G67R-intFLAG IPs More about this image found in Vid27 and Alx1 interact and promote NE assembly, independent of ESCRT-III. ...

Figure 6. AlphaFold2 prediction and biochemical validation of a Vid27 and Alx1 complex. (A) Interaction between the Vid27 PH2 domain (green surface) and C terminus of Alx1 (magenta sticks) predicted by AlphaFold2. (B and C) Confidence metrics More about this image found in AlphaFold2 prediction and biochemical validation of a Vid27 and Alx1 comple...

Figure 7. Alx1’s interaction with Vid27 is required for its NE assembly function. (A and B) Mutations at the predicted Alx1–Vid27 binding interface disrupt the Alx1-E308V interaction with Vid27. (A) IP of Vid27 or Vid27 binding surface mutants More about this image found in Alx1’s interaction with Vid27 is required for its NE assembly function. (A ...

Figure 1. Physiological UPR activation promotes meiosis in yeast. (A) Only the first of two Hac1 waves is required for meiotic entry. (B) Rather than inducing the full UPR regulon, the premeiotic UPR represses ribosome biogenesis and More about this image found in Physiological UPR activation promotes meiosis in yeast. (A) Only the first...

Figure 1. Comparison of two Golgiphagy reporter strategies. (A) Schematic representation of three Golgiphagy reporters Golgi5-297 (human, 633–731), B4GALT1 (human, 1–82 aa), Man2a1 (mouse, 1–116 aa) include the tandem fluorescent proteins More about this image found in Comparison of two Golgiphagy reporter strategies. (A) Schematic representa...