Telomeres and centromeres have interchangeable roles in promoting meiotic spindle formation

Files in this Data Supplement:

• Supplemental Material (PDF)
• Video 5 -
  Loss of nuclear movement results in centromere–SPB contact throughout and bipolar spindle formation. A bqt1Δ dhc1Δ meiocyte (h⁹⁰ Atb2-mCherry Sid4-mCherry Hht1-CFP Mis6-GFP bqt1Δ dhc1Δ) was imaged by time-lapse microscopy. The microscope settings used are described in Video 1.
• Video 3 -
  In the majority of cases, chromatin–SPB contact is mediated by the centromere. A bqt1Δ meiocyte (h⁹⁰ Atb2-mCherry Sid4-mCherry Hht1-CFP Mis6-GFP bqt1Δ) was imaged by time-lapse microscopy at 27°C in an Environmental Chamber with a DeltaVision Spectris (Applied Precision) as described in Video 1.
• MATLAB (zipped file) -
  MATLAB script for vector diagram of the range of contact lengths in individual cells shown in Fig. S1.
• Video 1 -
  bqt1Δ cells with no contact show defective spindle formation. A bqt1Δ meiocyte harboring GFP-tagged tubulin and SPB and mRFP-tagged histone H3 (h⁹⁰ Atb2-GFP Sid4-GFP Hht1-mRFP bqt1Δ) was imaged by time-lapse microscopy at 27°C in an Environmental Chamber with a DeltaVision Spectris (Applied Precision) comprising a widefield inverted epifluorescence microscope (IX70; Olympus), a UPlanSapo 100× NA 1.4 oil immersion objective (Olympus), and a charge coupled device CoolSnap HQ camera (Photometrics). Images were acquired over 26 focal planes at a 0.35-µm step size with frames taken every 10 min for 7 h.
• Video 2 -
  bqt1Δ cells with persistent (>30 min) chromatin–SPB contact show bipolar spindle formation. A bqt1Δ meiocyte (h⁹⁰ Atb2-GFP Sid4-GFP Hht1-mRFP bqt1Δ) was imaged by time-lapse microscopy at 27°C in an Environmental Chamber with a DeltaVision Spectris (Applied Precision) using the same settings as in Video 1.
• Video 4 -
  Artificial maintenance of centromere–SPB contacts ensures bipolar spindle formation. A rap1Δ lig4Δ meiocyte (h⁹⁰ Atb2-mCherry Sid4-mCherry Hht1-CFP Mis6-GFP rap1Δ lig4Δ Bqt1-GBP) was imaged by time-lapse microscopy at 27°C as described in Video 1.