Table I.
Effect of DOA4 over-expression on CPY-Inv secretion and GFP-CPS sorting
| Strain | −/+ 2 µ DOA4 a | Percentage of secreted CPY-Invb | GFP-CPS localization |
|---|---|---|---|
| Wild-type | − + |
1.0 ± 2.1c (5)d
2.8 ± 1.5 (4) |
VLe
VL |
| bro1 ΔCC | − + |
11.5 ± 4.7 (24) 3.7 ± 2.5 (7) |
VM + ECf
VL |
| bro1Δ | − + |
13.0 ± 3.8 (6) 5.6 ± 2.4 (5) |
VM + EC VL + VM + EC |
| vps4Δ | − + |
39.4 ± 2.5 (3) 5.2 ± 1.7 (3) |
VM + EC VM + EC |
| bro1Δ vps4Δ | − + |
43.7 ± 6.9 (3) 46.0 ± 7.1 (3 |
VM + EC VM + EC |
| bro1 ΔCC vps4Δ | − + |
35.2 ± 2.9 (3) 35.9 ± 4.1 (3) |
VM + EC VM + EC |
| ESCRT-I mutants | |||
| vps23Δ | − + |
20.0 ± 5.9 (3) 17.6 ± 2.0 (3) |
VM + EC VM + EC |
| vps28Δ | − + |
13.4 ± 1.2 (3) 21.7 ± 4.2 (3) |
VM + EC VM + EC |
| vps37Δ | − + |
16.3 ± 1.7 (3) 15.8 ± 1.3 (3) |
VLg
VL |
| ESCRT-II mutants | |||
| vps22Δ | − + |
20.1 ± 4.5 (3) 20.5 ± 6.1 (3) |
VM + EC VM + EC |
| vps25Δ | − + |
23.1 ± 1.0 (3) 20.7 ± 0.9 (3) |
VM + EC VM + EC |
| vps36Δ | − + |
24.2 ± 6.3 (3) 20.6 ± 4.1 (3) |
VM + EC VM + EC |
| ESCRT-III mutants | |||
| vps2Δ | − + |
25.2 ± 5.0 (3) 27.8 ± 6.5 (3) |
VM + EC VM + EC |
| vps24Δ | − + |
29.1 ± 4.3 (3) 30.9 ± 4.4 (3) |
VM + EC VM + EC |
| vps20Δ | − + |
24.2 ± 2.9 (3) 20.1 ± 6.5 (3) |
VM + EC VM + EC |
| snf7Δ | − + |
25.8 ± 5.0 (3) 24.6 ± 1.1 (3) |
VM + EC VM + EC |
| Other class E vps mutants tested | |||
| vps27Δ | − + |
28.0 ± 1.5 (3) 26.5 ± 2.6 (3) |
VM + EC VM + EC |
| nhx1Δ | − + |
41.8 ± 13.0 (3) 39.1 ± 13.0 (3) |
VLg
VL |
| vps60Δ | − + |
17.7 ± 1.3 (3) 19.0 ± 5.5 (3) |
VM + EC VM + EC |
a
Strains were transformed with the empty 2 µ vector (−) or the 2 µ vector containing DOA4 (+).
b
The percentage of total cellular CPY-Inv secreted by cells.
c
Mean ± standard error.
d
Number of independent experiments.
e
Vacuole lumen.
f
Vacuole membrane and class E compartment.
g
There was no noticeable defect in GFP-CPS sorting in vps37Δ and nhx1Δ cells.