Table I. Effects of various mutations on polarized recruitment of Bee1p or Vrp1p in the presence of Lat-A.
| Protein examined | Strain background | Cells with polarized protein |
|---|---|---|
| % | ||
| Bee1p-GFP | Wild-type | 47 ± 7 |
| cdc42A118 | 7 ± 4 | |
| cdc24-1 (24°C) | 44 ± 4 | |
| cdc24-1 (37°C) | 5 ± 2 | |
| CDC42L61 | 46* or 32 ± 14 | |
| vrp1ΔC | 5 ± 1 | |
| Δste20 cla4ts-degron (37°C) | 46 ± 8 | |
| Δbnr1 bni1ts (37°C) | 6 ± 3 | |
| Wild-type (+ α-factor) | 95 ± 3 | |
| Δbni1 (+ α-factor) | 9 ± 4 | |
| Δbnr1 bni1ts (+ α-factor, 24°C) | 56 ± 7 | |
| Δbnr1 bni1ts (+ α-factor, 37°C) | 6 ± 3 | |
| Vrp1p-GFP | Wild-type | 36 ± 3 |
| bee1ΔWH1 | 33 ± 4 |
Unless indicated with “+ α-factor,” polarization of Bee1p or Vrp1p was examined after release from a G1 arrest as described in Materials and methods. Localization of Bee1p or Vrp1p was determined by Immunofluorescence staining using an anti-GFP antibody.
*
Induction of polarization by CDC42L61 in G1-arrested cells (cdc28-13) was highly variable with values from 18 to 46% of cells showing Bee1p-GFP polarization. However, >80% of cells, which had detectable polarized CDC42L61-HA by immunofluorescence, showed polarized colocalization of Bee1p.