Table 3.
Mutants in genes involved in vacuolar protein sorting assayed for ADCB sensitivity
| Gene/parental strain namea | Proposed function in the literature/databases | Growth of deletion mutant on ADCBb |
|---|---|---|
| BY4741 | Wild type | + |
| CLASS A | ||
| PEP1/VPS10 | Sorting receptor for soluble vacuolar proteins | +/− |
| VPS13 | Late endosome to vacuole transport | + |
| VPS29 | Retromer complex (retrograde transport endosome to Golgi) | + |
| VPS30 | Sorting of soluble hydrolases to vacuole; required for autophagy | + |
| VPS35 | Retromer complex (retrograde transport endosome to Golgi) | + |
| VPS38 | Mediates interaction between Vps30 and Vps34p-Vps15p; late endosome to vacuole transport | + |
| VPS63 | Dubious ORF, 98% overlaps the gene YPT6; involved in CPY sorting/trafficking | + |
| VPS74 | CPY and Pep4p trafficking to the vacuole; apical bud growth | + |
| CLASS B | ||
| VAM3 | Syntaxin homolog (t-SNARE), required for vacuolar assembly | + |
| VPS5 | Retromer complex (retrograde transport endosome to Golgi) | + |
| VPS17 | Retromer complex (retrograde transport endosome to Golgi) | + |
| VAM6/VPS39 | HOPS complex | + |
| VPS41 | HOPS complex | + |
| VAM7/VPS43 | Vacuolar SNARE complex | + |
| VPS52 | Subunit of the VFT/GARP complex, involved in retrograde transport, endosome to Golgi | + |
| VPS53 | Subunit of the VFT/GARP complex, involved in retrograde transport, endosome to Golgi | + |
| VPS54 | Subunit of the VFT/GARP complex, involved in retrograde transport, endosome to Golgi | + |
| VPS51/VPS67 | Subunit of the VFT/GARP complex, involved in retrograde transport, endosome to Golgi | + |
| VPS61 | Deletion causes defects in CPY and Pho8p trafficking to the vacuole | + |
| VPS64/FAR9 | CPY trafficking and α-factor secretion | + |
| VPS66 | CPY vacuolar trafficking pathway | − |
| VPS69 | CPY vacuolar trafficking pathway | +/− |
| VPS71 | Component of the Swr1p complex; CPY vacuolar trafficking pathway | − |
| VPS72 | Component of the Swr1p complex; CPY vacuolar trafficking pathway | − |
| VPS73 | Mitochondrial protein of unknown function involved in vacuolar protein sorting | + |
| VPS75 | CPY and Pho8p trafficking to the vacuole | + |
| CLASS C | ||
| PEP5/VPS11 | HOPS complex | + |
| VPS16 | HOPS complex | +/− |
| PEP3/VPS18 | HOPS complex | − |
| VPS33 | HOPS complex | + |
| CLASS D | ||
| VPS3 | Vacuolar sorting and segregation | − |
| VPS8 | Retrieval of Golgi membrane proteins from prevacuolar compartment | + |
| PEP12/VPS6 | Syntaxin homolog (t-SNARE) involved in Golgi to vacuole transport | − |
| VPS9 | Required for Golgi to vacuole transport | + |
| VPS15 | Myristoylated Ser/Thr protein kinase possibly involved in the activation of Vps34p | + |
| PEP7/VPS19 | Vacuolar segregation and acidification | − |
| VPS21 | Rab5-like GTPase required for sorting of vacuolar proteins | + |
| VPS34 | Phosphatidyl inositol (PI) 3-kinase required for vacuolar protein sorting | + |
| VPS45 | Protein of the Sec1p family, essential for vacuolar protein sorting | + |
| Class E | ||
| DID2/FTI1 | Small coiled-coil protein; human ortholog may be altered in breast tumors | +/− |
| VPS60/MOS10 | Small coiled-coil protein with notable identity to VPS32 and VPS20 | +/− |
| HSE1 | Sorting of ubiquitinated cargo into MVEs; interacts with Vps27p | + |
| VPS27 | Sorting of ubiquitinated cargo into MVEs; recruits/activates ESCRT-I on endosomes | − |
| STP22/VPS23 | ESCRT-I complex component (MVE sorting pathway) | − |
| VPS28 | ESCRT-I complex component (MVE sorting pathway) | − |
| SRN2/VPS37 | ESCRT-I complex component (MVE sorting pathway) | − |
| SNF8/VPS22 | ESCRT-II complex component (MVE sorting pathway) | − |
| VPS25 | ESCRT-II complex component (MVE sorting pathway) | − |
| VPS36 | ESCRT-II complex component (MVE sorting pathway) | − |
| SNF7/VPS32 | ESCRT-III complex component (subcomplex with Vps20p; MVE sorting pathway) | − |
| VPS20 | ESCRT-III complex component (subcomplex with Vps32p; MVE sorting pathway) | − |
| DID4/VPS2 | ESCRT-III complex component (MVE sorting pathway) | − |
| VPS24 | ESCRT-III complex component (subcomplex with Vps2p) | − |
| VPS4 | AAA ATPase protein; release of ESCRT complexes from endosomal membranes | − |
| BRO1/VPS31 | Necessary for efficient recruitment of Doa4p into endosomes | − |
| NHX1/VPS44 | Na+/H+ antiporter required for endosomal trafficking | − |
| Class F | ||
| VPS1 | GTPase; vacuolar sorting and retention of Golgi proteins | + |
| PEP8/VPS26 | Retromer complex (retrograde transport endosome to Golgi) | + |
| VPS55 | Late endosome to vacuole trafficking | + |
| VPS62 | CPY and Pep4p trafficking to the vacuole | + |
| VPS65 | Dubious ORF; vacuolar protein sorting; actin organization | − |
| VPS68 | CPY and Pep4p trafficking to the vacuole | + |
| VPS70 | Vacuolar protein sorting; protease associated domain | + |
| Others | ||
| FAB1 | PI(3)P-5 kinase involved in MVE formation | + |
| DEF1/VID31 | Protein involved in vacuolar import and degradation | +/− |
| DOA4 | Ub C-terminal hydrolase; recycling of Ub from protein substrates | − |
| TUL1 | Transmembrane Ub-ligase involved in the ubiquitination of misfolded membrane proteins | + |
| PIB1 | E2-dependent Ub ligase resident of endosomal and vacuolar membranes | + |
a Genes are listed according to the Saccharomyces Genome Database (the most common alias has also been included for some of them).
b Growth is represented according to the survival of the mutant strains on plates of minimal urea medium with different concentrations of ADCB: +, mutants with wild type behavior, unable to grow in concentrations above 30 mg/l; +/−, moderately sensitive mutants unable to grow in concentrations above 12.5 mg/l; −, hypersensitive mutants unable to grow above 7 mg/l. Sublethal concentrations for the control strains were 6.25 mg/l for a gln3Δ mutant and >100 mg/l for an lst4Δ mutant.