Table 4.
Effect on Gap1p activity of vacuolar sorting mutants selected in the genomic screen
| Gene | Proposed function for the encoded protein | Growth on ADCBa | Gap1p activity (%)b | Arginine uptake (%) |
|---|---|---|---|---|
| BY4741 | Wild type | + | 100 ± 13 | 100 ± 04 |
| CLASS A | ||||
| VPS29 | Retromer complex | + | 436 ± 103 | 82 ± 11 |
| VPS35 | Retromer complex | + | 309 ± 51 | 107 ± 01 |
| CLASS B | ||||
| VAM3 | Homotypic vacuolar fusion | + | 327 ± 51 | 111 ± 13 |
| VPS5 | Retromer complex | + | 418 ± 121 | 121 ± 23 |
| VPS17 | Retromer complex | + | 73 ± 13 | 97 ± 04 |
| VPS52 | VFT/GARP complex | + | 336 ± 39 | 70 ± 03 |
| VPS53 | VFT/GARP complex | + | 345 ± 26 | 68 ± 05 |
| VPS54 | VFT/GARP complex | + | 340 ± 16 | 65 ± 10 |
| VPS66 | Vacuolar trafficking | − | 482 ± 39 | 56 ± 23 |
| VPS69 | Vacuolar trafficking | − | 585 ± 95 | 89 ± 37 |
| VPS71 | Vacuolar trafficking | − | 394 ± 165 | 156 ± 96 |
| CLASS C | ||||
| VPS16 | HOPS complex | +/− | 894 ± 181 | 71 ± 11 |
| PEP3/VPS18 | HOPS complex | − | 1000 ± 178 | 125 ± 64 |
| CLASS D | ||||
| VPS3 | Vacuolar segregation | − | 2862 ± 105 | 184 ± 16 |
| PEP12 | t-SNARE of the PVC | − | 973 ± 39 | 98 ± 14 |
| VPS9 | GDP/GTP exchange factor | − | 372 ± 105 | 95 ± 05 |
| VPS19 | Vesicular transport from Golgi to endosomes | − | 536 ± 17 | 140 ± 06 |
| VPS45 | Activation of t-SNARES in vesicular | + | 518 ± 116 | 72 ± 13 |
| transport from Golgi to endosomes | ||||
| CLASS E | ||||
| DID2/FTI1 | Endosomal coiled-coil protein | +/− | 582 ± 257 | 127 ± 32 |
| VPS60/MOS10 | Endosomal coiled-coil protein | +/− | 1082 ± 501 | 137 ± 38 |
| HSE1 | In complex with Vps27p | + | 682 ± 64 | 185 ± 90 |
| VPS27 | Recognition of cargo at the MVE | − | 1938 ± 985 | 132 ± 75 |
| STP22/VPS23 | ESCRT-I complex | − | 627 ± 270 | 89 ± 52 |
| VPS28 | ESCRT-I complex | − | 1118 ± 656 | 143 ± 50 |
| SRN2/VPS37 | ESCRT-I complex | − | 764 ± 309 | 123 ± 13 |
| SNF8/VPS22 | ESCRT-II complex | − | 973 ± 107 | 107 ± 42 |
| VPS25 | ESCRT-II complex | − | 2098 ± 302 | 136 ± 04 |
| VPS36 | ESCRT-II complex | − | 1164 ± 129 | 136 ± 09 |
| VPS20 | ESCRT-III complex | − | 973 ± 64 | 105 ± 46 |
| SNF7/VPS32 | ESCRT-III complex | − | 964 ± 77 | 88 ± 48 |
| DID4/VPS2 | ESCRT-III complex | − | 2863 ± 425 | 137 ± 69 |
| VPS24 | ESCRT-III complex | − | 2682 ± 119 | 149 ± 14 |
| VPS4 | Disassembly of complexes at the MVE | − | 1902 ± 720 | 135 ± 36 |
| BRO1/VPS31 | Doa4p recruitment to MVE | − | 2787 ± 437 | 187 ± 25 |
| NHX1 | Na+/H+ antiporter | − | 553 ± 90 | 92 ± 12 |
| CLASS F | ||||
| PEP8/VPS26 | Retromer complex | + | 427 ± 116 | 99 ± 31 |
| VPS65 | Cytoskeleton organization | − | 521 ± 45 | 75 ± 01 |
| OTHERS | ||||
| FAB1 | PI(3)P-5 kinase; MVEs function | + | 100 ± 13 | 84 ± 19 |
| DEF1/VID31 | Vacuolar sorting | + | 265 ± 20 | 70 ± 30 |
| DOA4 | Deubiquitination of cargo at the MVEs | − | 2560 ± 132 | 119 ± 08 |
| TUL1 | Ubiquitination of misfolded proteins targeted to vacuole for degradation | + | 309 ± 51 | 113 ± 16 |
| PIB1 | Ub ligase, endosomal/vacuolar resident | + | 100 ± 13 | 127 ± 22 |
a ADCB phenotype is expressed as in Table 3.
b Gap1p activity in each mutant is expressed as a percentage of the [14C]citrulline uptake rate of wild type cells growing in minimal urea liquid medium. [14C]arginine uptake is also shown as a control. The SD of four independent experiments is shown.