Plasmids used
| Plasmids | Description | Use | Source |
| pRS315pRS316 | Centromere containing low copy yeast shuttle plasmids (LEU2 and URA3) | Figs. 1 (A, C, and, D), 2 A, 5 (A–C), and S2 A | Sikorski and Hieter, 1989 |
| pAC376 | Yep351 (2µ LEU2) expressing myc-tagged Ub from CUP1 promoter | Figs. 1 (A and C), 2 A, and 4 B | A. Cooper (Garvan Institute, Sydney Australia) |
| pRSmga2NΔtm | HIS3 low copy plasmid containing a truncated MGA2 gene lacking C-terminal 329 codons that include the membrane-spanning domain. | Figs. 2 D and S1 D | Chellappa et al., 2001 |
| pYM-N36 | nourseothricin-resistance marker natMX | Janke et al., 2004 | |
| Genomic DNA | HSV-1 (strain F) | Ejercito et al., 1968 | |
| pPL4149 | pRS316 expressing HA-Vps27 from MET25 promoter | Figs. 1 (A, C, and D), 2 A, 4 B, and S1 (A and B) | This study |
| pPL4150 | pRS316 expressing HA-Vps27K>R (where all K residues are altered to R) from MET25 promoter. | Figs. 1 (C and D) and S1 (A and B) | This study |
| pPL967 | pRS315 expressing Ste3-GFP from STE3 promoter | Figs. 1 D, 2 D, 3 (A and E), 4 C, 5 (B and D), and S2 F | Urbanowski and Piper, 2001 |
| pPL991 | pRS313 expressing Ste3-GFP from STE3 promoter | Fig. 1 D | Urbanowski and Piper, 2001 |
| pPL2334 | pRS315 expressing Gap1-GFP from CUP1 promoter | Figs. 2 (C and D), 3 (A and B), 4 C, and 5 (C andD) | This study |
| pPL3484 | pRS315 expressing Ste3-GFP-Ub from STE3 promoter | Figs. 4 C and 5, A, B, and D | This study |
| pPL3797 | pRS315 expressing Fur4-GFP from CUP1 promoter | Figs. 3 A, 4 C, and 5, A and D | This study |
| pPL3931 | ADE2 conversion of pPL3484 (ADE2 replacement of LEU2) expressing Ste3-GFP-Ub from STE3 promoter | Fig. 2 D | This study |
| pPL3962 | pRS315 expressing Hxt1-GFP from HXT1 promoter | Fig. 3 A | This study |
| pPL4144 | pRS315 expressing Fur4-mCherry-Ub from CUP1 promoter | Fig. 5 C | This study |
| pPL4146 | pRS315 expressing Mup1-GFP from CUP1 promoter | Figs. 2 (B, C, and E), 4 C, and 5D | This study |
| pPL4147 | pRS315 expressing Mup1-GFP-Ub from CUP1 promoter | Figs. 2 (B–D), 4 C, 5 D, and S2 F | This study |
| pPL4214 | ADE2 conversion of pPL967 (ADE2 replacement of LEU2) expressing Ste3-GFP from STE3 promoter | Fig. 2 D | This study |
| pPL4215 | ADE2 conversion of pPL2334 (ADE2 replacement of LEU2) expressing Gap1-GFP from CUP1 promoter | Fig. 2 D | This study |
| pPL4216 | ADE2 conversion of pPL4146 (ADE2 replacement of LEU2) expressing Mup1-GFP from CUP1 promoter | Fig. 2 D | This study |
| pPL4217 | ADE2 conversion of pPL4147 (ADE2 replacement of LEU2) expressing Mup1-GFP-Ub from CUP1 promoter | Fig. 2 D | This study |
| pPL4252 | natMX conversion of pPL4146 (natMX replacement of LEU2) expressing Mup1-GFP from CUP1 promoter | Figs. 2 E and S2 F | This study |
| pPL4320 | pRS315 expressing Mup1-GFP-UbK63R from CUP1 promoter | This study | |
| pPL4342 | natMX conversion of pPL4320 (natMX replacement of LEU2) expressing Mup1-GFP-UbK63R from CUP1 promoter | Fig. 2 E | This study |
| pPL3584 | pRS316 expressing Mvb12-Ubp7CD-3xHA from CUP1 promoter; contains residues 561-1071 of Ubp7 | Figs. 4 C, 5 A, and S1 F | This study |
| pPL3609 | pRS316 expressing Ste3-mCherry-Ubp7CD from STE3 promoter | Fig. 3 B | This study |
| pPL3669 | pRS316 expressing Fur4-GFP-Ubp7CD from CUP1 promoter | Fig. 3 B | This study |
| pPL3810 | pRS316 expressing Fur4-mCherry-Ubp7CD from CUP1 promoter | Fig. 3 B | This study |
| pPL3876 | pRS316 expressing Ste3-GFP-Ubp7CD from STE3 promoter | Figs. 3 A and S1 E | This study |
| pPL3900 | pRS316 expressing Gap1-GFP-Ubp7CD from CUP1 promoter | Fig. 3 A | This study |
| pPL3964 | pRS316 expressing Hxt1-GFP-Ubp7CD from HXT1 promoter | Fig. 3 A | This study |
| pPL3986 | pRS316 expressing Ste3-GFP-Ubp7CD(G>S) -3xHA from STE3 promoter; contains residues 561–1,071 of Ubp7 with mutations G932S | Figs. S1 E and S2B | This study |
| pPL3890 | pRS316 expressing Ste3-GFP-3xHA from STE3 promoter | Fig. S1 E | This study |
| pPL3722 | pRS316 expressing His3-Ubp7CD-3xHA from CUP1 promoter. Contains residues 561–1071 of Ubp7 followed by 3×HA epitope | Figs. 4C, S1 F, and S2, A and C | This study |
| pPL3723 | pRS316 expressing Hse1-Ubp7CD-3xHA from CUP1 promoter | Figs. 4 C, S1 F, and S2 A | This study |
| pPL3724 | pRS316 expressing Vps23-Ubp7CD-3xHA from CUP1 promoter | Figs. 4 C, S1 F, and S2 A | This study |
| pPL3742 | pRS316 expressing Rsp5-Ubp7CD-3xHA from CUP1 promoter | Figs. 5 D, and S2 B | This study |
| pPL3746 | pRS316 expressing Gga2-Ubp7CD-3xHA from CUP1 promoter | Figs. 4 C, S1 F, and S2 A | This study |
| pPL3643 | pRS316 expressing Ste3-GFP-ubp7CD* from STE3 promoter; contains residues 561–1071 of Ubp7 with C618S mutation | Fig. 3 A | This study |
| pPL3776 | pRS316 expressing Mvb12- ubp7CD*-3xHA from CUP1 promoter | Figs. 4 C, 5 A, S1 F, and S2 A | This study |
| pPL3901 | pRS316 expressing Gap1-GFP- ubp7CD* from CUP1 promoter | Fig. 3 A | This study |
| pPL4371 | pRS316 expressing Hxt1-GFP-ubp7CD* from HXT1 promoter | Fig. 3 A | This study |
| pPL3994 | pRS316 expressing Ste3-GFP-UL36CD from STE3 promoter; contains 561–591 of Ubp7 (Ubp7 linker)-and residues 15–260 of HSV1 UL36 | Fig. 3, A and D | This study |
| pPL4012 | pRS316 expressing Hse1-UL36CD-3xHA from CUP1 promoter | Figs. 4 (B and C), 5 B, S1 G, and S2 A | This study |
| pPL4013 | pRS316 expressing Mvb12-UL36CD-3xHA from CUP1 promoter | Figs. 4 C, 5 A, S1 G, and S2 A | This study |
| pPL4014 | pRS316 expressing His3-UL36CD-3xHA from CUP1 promoter | Figs. 4 C, S1 G, and S2 A | This study |
| pPL4015 | pRS316 expressing Vps23-GFP-UL36CD from VPS23 promoter | Fig. 5 C | This study |
| pPL4017 | pRS316 expressing Gga2-UL36CD-3xHA from CUP1 promoter | Figs. 4 C, S1 G, and S2 A | This study |
| pPL4140 | HIS3 conversion of pPL4012 expressing Hse1-UL36CD-3xHA from CUP1 promoter | Fig. 4 B | This study |
| pPL4247 | pRS316 expressing Vps23-UL36CD-3xHA from CUP1 promoter | Figs. 4 and S2, B and C | This study |
| pPL4058 | pRS316 expressing Ste3-GFP-ul36CD* from STE3 promoter; contains 561–591 of Ubp7 (Ubp7 linker) and residues 15–260 of HSV1 UL36 with mutation of C40S in UL36 | Fig. 3, B and D | This study |
| pPL4131 | pRS316 expressing Hse1-ul36CD*-3xHA from CUP1 promoter | Figs. 4 C, 5 B, and S2, B and C | This study |
| pPL4141 | HIS3 conversion of pPL4131 expressing Hse1-ul36CD*-3xHA from CUP1 promoter | Fig. 4 C | This study |
| pJLU40 | pRS316 expressing Vph1-GFP from VPH1 promoter | Fig. 3 A | This study |
| pPL4056 | pRS316 expressing Vph1-GFP- UL36CD from VPH1 promoter | Fig. 3 A | This study |
| pPL4202 | pRS316 expressing Vph1-GFP- ul36CD* from VPH1 promoter | Fig. 3 A | This study |
| pPL4218 | pRS316 expressing Ste3-GFP-M48CD from STE3 promoter; contains 561–591 of Ubp7 (Ubp7 linker) AGDRDIV-mCMV M48 (residues 4–234)-VAITHPLR. | Fig. 3 A | This study |
| pPL4219 | pRS316 expressing Ste3-GFP-m48CD* from STE3 promoter; contains linker residues 561–591 of Ubp7, residues 15–21 of UL36, residues 4–234 of mCMV M48, and the mutations C23A, L110A, Y113A, and Y192A followed by residues (253–260 of UL36) | Fig. 3 A | This study |
| pPL3974 | pET151D expressing UL36CD (residues 15–260 of HSV1 UL36) | Fig. S2 E | This study |
| pPL2554 | HIS3 conversion of pPL2356 (HIS3 replacement of URA3) expressing GFP-Cps1 from Cps1 promoter | Fig. S1 B | This study |
| pPL4500 | pRS316 expressing Ste3-GFP-M48-3xHA from STE3 promoter | Fig. 3 C | This study |
| pPL4502 | pRS316 expressing Rsp5-ubp7*-3xHA from CUP1 promoter | Figs. 5 D and S2 B | This study |
| pPL4513 | pRS316 expressing Ste3-GFP-m48*-3xHA from STE3 promoter | Fig. 3 C | This study |
| pPL4515 | pRS16 expressing Ste3-GFP-ubp7*-3xHA from STE3 promoter | Fig. S1 E | This study |
| pPL4253 | natMX conversion of pPL4147 (natMX replacement of LEU2) expressing Mup1-GFP-Ub from CUP1 promoter | Fig. S2 F | This study |
| pPL4248 | natMX conversion of pPL967 (natMX replacement of LEU2) expressing Ste3-GFP from STE3 promoter | Fig. S2 F | This study |
| pPL4249 | natMX conversion of pPL3484 (natMX replacement of LEU2) expressing Ste3-GFP-Ub from STE3 promoter | Fig. S2 F | This study |
| Plasmids | Description | Use | Source |
| pRS315pRS316 | Centromere containing low copy yeast shuttle plasmids (LEU2 and URA3) | Figs. 1 (A, C, and, D), 2 A, 5 (A–C), and S2 A | Sikorski and Hieter, 1989 |
| pAC376 | Yep351 (2µ LEU2) expressing myc-tagged Ub from CUP1 promoter | Figs. 1 (A and C), 2 A, and 4 B | A. Cooper (Garvan Institute, Sydney Australia) |
| pRSmga2NΔtm | HIS3 low copy plasmid containing a truncated MGA2 gene lacking C-terminal 329 codons that include the membrane-spanning domain. | Figs. 2 D and S1 D | Chellappa et al., 2001 |
| pYM-N36 | nourseothricin-resistance marker natMX | Janke et al., 2004 | |
| Genomic DNA | HSV-1 (strain F) | Ejercito et al., 1968 | |
| pPL4149 | pRS316 expressing HA-Vps27 from MET25 promoter | Figs. 1 (A, C, and D), 2 A, 4 B, and S1 (A and B) | This study |
| pPL4150 | pRS316 expressing HA-Vps27K>R (where all K residues are altered to R) from MET25 promoter. | Figs. 1 (C and D) and S1 (A and B) | This study |
| pPL967 | pRS315 expressing Ste3-GFP from STE3 promoter | Figs. 1 D, 2 D, 3 (A and E), 4 C, 5 (B and D), and S2 F | Urbanowski and Piper, 2001 |
| pPL991 | pRS313 expressing Ste3-GFP from STE3 promoter | Fig. 1 D | Urbanowski and Piper, 2001 |
| pPL2334 | pRS315 expressing Gap1-GFP from CUP1 promoter | Figs. 2 (C and D), 3 (A and B), 4 C, and 5 (C andD) | This study |
| pPL3484 | pRS315 expressing Ste3-GFP-Ub from STE3 promoter | Figs. 4 C and 5, A, B, and D | This study |
| pPL3797 | pRS315 expressing Fur4-GFP from CUP1 promoter | Figs. 3 A, 4 C, and 5, A and D | This study |
| pPL3931 | ADE2 conversion of pPL3484 (ADE2 replacement of LEU2) expressing Ste3-GFP-Ub from STE3 promoter | Fig. 2 D | This study |
| pPL3962 | pRS315 expressing Hxt1-GFP from HXT1 promoter | Fig. 3 A | This study |
| pPL4144 | pRS315 expressing Fur4-mCherry-Ub from CUP1 promoter | Fig. 5 C | This study |
| pPL4146 | pRS315 expressing Mup1-GFP from CUP1 promoter | Figs. 2 (B, C, and E), 4 C, and 5D | This study |
| pPL4147 | pRS315 expressing Mup1-GFP-Ub from CUP1 promoter | Figs. 2 (B–D), 4 C, 5 D, and S2 F | This study |
| pPL4214 | ADE2 conversion of pPL967 (ADE2 replacement of LEU2) expressing Ste3-GFP from STE3 promoter | Fig. 2 D | This study |
| pPL4215 | ADE2 conversion of pPL2334 (ADE2 replacement of LEU2) expressing Gap1-GFP from CUP1 promoter | Fig. 2 D | This study |
| pPL4216 | ADE2 conversion of pPL4146 (ADE2 replacement of LEU2) expressing Mup1-GFP from CUP1 promoter | Fig. 2 D | This study |
| pPL4217 | ADE2 conversion of pPL4147 (ADE2 replacement of LEU2) expressing Mup1-GFP-Ub from CUP1 promoter | Fig. 2 D | This study |
| pPL4252 | natMX conversion of pPL4146 (natMX replacement of LEU2) expressing Mup1-GFP from CUP1 promoter | Figs. 2 E and S2 F | This study |
| pPL4320 | pRS315 expressing Mup1-GFP-UbK63R from CUP1 promoter | This study | |
| pPL4342 | natMX conversion of pPL4320 (natMX replacement of LEU2) expressing Mup1-GFP-UbK63R from CUP1 promoter | Fig. 2 E | This study |
| pPL3584 | pRS316 expressing Mvb12-Ubp7CD-3xHA from CUP1 promoter; contains residues 561-1071 of Ubp7 | Figs. 4 C, 5 A, and S1 F | This study |
| pPL3609 | pRS316 expressing Ste3-mCherry-Ubp7CD from STE3 promoter | Fig. 3 B | This study |
| pPL3669 | pRS316 expressing Fur4-GFP-Ubp7CD from CUP1 promoter | Fig. 3 B | This study |
| pPL3810 | pRS316 expressing Fur4-mCherry-Ubp7CD from CUP1 promoter | Fig. 3 B | This study |
| pPL3876 | pRS316 expressing Ste3-GFP-Ubp7CD from STE3 promoter | Figs. 3 A and S1 E | This study |
| pPL3900 | pRS316 expressing Gap1-GFP-Ubp7CD from CUP1 promoter | Fig. 3 A | This study |
| pPL3964 | pRS316 expressing Hxt1-GFP-Ubp7CD from HXT1 promoter | Fig. 3 A | This study |
| pPL3986 | pRS316 expressing Ste3-GFP-Ubp7CD(G>S) -3xHA from STE3 promoter; contains residues 561–1,071 of Ubp7 with mutations G932S | Figs. S1 E and S2B | This study |
| pPL3890 | pRS316 expressing Ste3-GFP-3xHA from STE3 promoter | Fig. S1 E | This study |
| pPL3722 | pRS316 expressing His3-Ubp7CD-3xHA from CUP1 promoter. Contains residues 561–1071 of Ubp7 followed by 3×HA epitope | Figs. 4C, S1 F, and S2, A and C | This study |
| pPL3723 | pRS316 expressing Hse1-Ubp7CD-3xHA from CUP1 promoter | Figs. 4 C, S1 F, and S2 A | This study |
| pPL3724 | pRS316 expressing Vps23-Ubp7CD-3xHA from CUP1 promoter | Figs. 4 C, S1 F, and S2 A | This study |
| pPL3742 | pRS316 expressing Rsp5-Ubp7CD-3xHA from CUP1 promoter | Figs. 5 D, and S2 B | This study |
| pPL3746 | pRS316 expressing Gga2-Ubp7CD-3xHA from CUP1 promoter | Figs. 4 C, S1 F, and S2 A | This study |
| pPL3643 | pRS316 expressing Ste3-GFP-ubp7CD* from STE3 promoter; contains residues 561–1071 of Ubp7 with C618S mutation | Fig. 3 A | This study |
| pPL3776 | pRS316 expressing Mvb12- ubp7CD*-3xHA from CUP1 promoter | Figs. 4 C, 5 A, S1 F, and S2 A | This study |
| pPL3901 | pRS316 expressing Gap1-GFP- ubp7CD* from CUP1 promoter | Fig. 3 A | This study |
| pPL4371 | pRS316 expressing Hxt1-GFP-ubp7CD* from HXT1 promoter | Fig. 3 A | This study |
| pPL3994 | pRS316 expressing Ste3-GFP-UL36CD from STE3 promoter; contains 561–591 of Ubp7 (Ubp7 linker)-and residues 15–260 of HSV1 UL36 | Fig. 3, A and D | This study |
| pPL4012 | pRS316 expressing Hse1-UL36CD-3xHA from CUP1 promoter | Figs. 4 (B and C), 5 B, S1 G, and S2 A | This study |
| pPL4013 | pRS316 expressing Mvb12-UL36CD-3xHA from CUP1 promoter | Figs. 4 C, 5 A, S1 G, and S2 A | This study |
| pPL4014 | pRS316 expressing His3-UL36CD-3xHA from CUP1 promoter | Figs. 4 C, S1 G, and S2 A | This study |
| pPL4015 | pRS316 expressing Vps23-GFP-UL36CD from VPS23 promoter | Fig. 5 C | This study |
| pPL4017 | pRS316 expressing Gga2-UL36CD-3xHA from CUP1 promoter | Figs. 4 C, S1 G, and S2 A | This study |
| pPL4140 | HIS3 conversion of pPL4012 expressing Hse1-UL36CD-3xHA from CUP1 promoter | Fig. 4 B | This study |
| pPL4247 | pRS316 expressing Vps23-UL36CD-3xHA from CUP1 promoter | Figs. 4 and S2, B and C | This study |
| pPL4058 | pRS316 expressing Ste3-GFP-ul36CD* from STE3 promoter; contains 561–591 of Ubp7 (Ubp7 linker) and residues 15–260 of HSV1 UL36 with mutation of C40S in UL36 | Fig. 3, B and D | This study |
| pPL4131 | pRS316 expressing Hse1-ul36CD*-3xHA from CUP1 promoter | Figs. 4 C, 5 B, and S2, B and C | This study |
| pPL4141 | HIS3 conversion of pPL4131 expressing Hse1-ul36CD*-3xHA from CUP1 promoter | Fig. 4 C | This study |
| pJLU40 | pRS316 expressing Vph1-GFP from VPH1 promoter | Fig. 3 A | This study |
| pPL4056 | pRS316 expressing Vph1-GFP- UL36CD from VPH1 promoter | Fig. 3 A | This study |
| pPL4202 | pRS316 expressing Vph1-GFP- ul36CD* from VPH1 promoter | Fig. 3 A | This study |
| pPL4218 | pRS316 expressing Ste3-GFP-M48CD from STE3 promoter; contains 561–591 of Ubp7 (Ubp7 linker) AGDRDIV-mCMV M48 (residues 4–234)-VAITHPLR. | Fig. 3 A | This study |
| pPL4219 | pRS316 expressing Ste3-GFP-m48CD* from STE3 promoter; contains linker residues 561–591 of Ubp7, residues 15–21 of UL36, residues 4–234 of mCMV M48, and the mutations C23A, L110A, Y113A, and Y192A followed by residues (253–260 of UL36) | Fig. 3 A | This study |
| pPL3974 | pET151D expressing UL36CD (residues 15–260 of HSV1 UL36) | Fig. S2 E | This study |
| pPL2554 | HIS3 conversion of pPL2356 (HIS3 replacement of URA3) expressing GFP-Cps1 from Cps1 promoter | Fig. S1 B | This study |
| pPL4500 | pRS316 expressing Ste3-GFP-M48-3xHA from STE3 promoter | Fig. 3 C | This study |
| pPL4502 | pRS316 expressing Rsp5-ubp7*-3xHA from CUP1 promoter | Figs. 5 D and S2 B | This study |
| pPL4513 | pRS316 expressing Ste3-GFP-m48*-3xHA from STE3 promoter | Fig. 3 C | This study |
| pPL4515 | pRS16 expressing Ste3-GFP-ubp7*-3xHA from STE3 promoter | Fig. S1 E | This study |
| pPL4253 | natMX conversion of pPL4147 (natMX replacement of LEU2) expressing Mup1-GFP-Ub from CUP1 promoter | Fig. S2 F | This study |
| pPL4248 | natMX conversion of pPL967 (natMX replacement of LEU2) expressing Ste3-GFP from STE3 promoter | Fig. S2 F | This study |
| pPL4249 | natMX conversion of pPL3484 (natMX replacement of LEU2) expressing Ste3-GFP-Ub from STE3 promoter | Fig. S2 F | This study |