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. 2020 May 26;21(7):e49929. doi: 10.15252/embr.201949929

Figure 6. Actin organization, but not microtubules, is essential for UapA translocation to the PM.

Figure 6

  • A
    Time course of treatment with the actin polymerization drug latrunculin B for 2, 5, 10, 30 or 50 min of a strain expressing neosynthesized UapA‐GFP under conditions of derepression compared to an untreated strain included as control (150 min). In all cases, latrunculin B was added at 100 min of UapA derepression, so that total time of UapA‐GFP expression was 102, 105, 110, 130 or 150 min in the different samples. Notice the abolishment of sorting of UapA to the PM after 130 or 150 min of expression when latrunculin B was present for the last 30 or 50 min, respectively. Scale bar: 5 μm.
  • B
    Time course of treatment of strains expressing neosynthesized alcA p‐UapA‐GFP and mCherry‐TubA with the anti‐microtubule drug benomyl for 5, 55 or 95 min. In all cases, benomyl was added at 95 min of UapA derepression, so that total time of UapA‐GFP expression was 100, 150 or 190 min. Benomyl abolished the thread‐like appearance of microtubules in all samples added, evident by the diffuse cytoplasmic signal of mCherry‐TubA. Notice that UapA reaches normally the PM (at 190 min of derepression), similarly to the untreated control (right panel). Scale bar: 5 μm.
  • C
    Subcellular localization of neosynthesized alcA p ‐GFP‐ChsB in the absence (left panel) or presence (≈40 min) of benomyl (middle panel) or latrunculin B (right panel), after ≈2 h of derepression. Notice the abolishment of proper polar localization of ChsB at the apical tip in both cases. Scale bars: 5 μm.
  • D
    Effect of latrunculin B or benomyl on COPII vesicle formation, followed through the subcellular localization Sec24‐GFP. Notice that 40 min of latrunculin B led to loss of the wild‐type punctuate appearance of Sec24 cytoplasmic puncta and the appearance of a diffuse fluorescent haze in the cytoplasm. In contrast, benomyl treatment did not affect the wild‐type punctuate localization of Sec24. Scale bar: 5 μm.

Source data are available online for this figure.