Table II.

Plasmids used in this study

Plasmid Description Source 
pPL2612 vps36ΔUb (T187F G187A) This study 
pPL2613 VPS36 (−309–1,698 VPS36 and 840 bp of the PHO8 3′UT) in pRS316 This study 
pPL2612 vps36ΔUb (T187F/G187A) in pRS316 This study 
pPL2645 vps36ΔUb (T187F/G187A) as a NotI–Acc65I fragment from pPL2612 in pRS304 This study 
pPL2646 VPS36 as a NotI–Acc65I fragment from pPL2613 in pRS304 This study 
pPL2940 vps23ΔUb1 in pRS306 (F52AQ117AW125A) This study 
pPL2180 VPS23 (−598–1,298) as an XbaI–SacI fragment in pRS316 with BamHI site removed from polylinker with an XbaI–XhoI fill-in ligation This study 
pPL2447 vps23 with UEV mutation F52A in pPL2180 This study 
pPL2448 vps23 with UEV mutation N123A in pPL2180 This study 
pPL2449 vps23 with UEV mutation W125A in pPL2180 This study 
pPL2451 vps23 with UEV mutation S55AD56A in pPL2180 This study 
pPL2452 vps23 with UEV mutation F52AS55AD56A in pPL2180 This study 
pPL2453 vps23 with UEV mutation S55AD56AN123A in pPL2180 This study 
pPL2465 vps23 with UEV mutation S55A in pPL2180 This study 
pPL2466 vps23 with UEV mutation S55AF105A in pPL2180 This study 
pPL2567 vps23 with UEV mutation F105AW125A in pPL2180 This study 
pPL2568 vps23 with UEV mutation F52AF105A in pPL2180 This study 
pPL2569 vps23 with UEV mutation F105A in pPL2180 This study 
pPL2570 vps23 with UEV mutation G57A in pPL2180 This study 
pPL2571 vps23 with UEV mutation G57AF105A in pPL2180 This study 
pPL2578 vps23 with UEV mutation F52AW125A in pPL2180 This study 
pPL2651 vps23 with UEV mutation F52AN123AW125A in pPL2180 This study 
pPL2652 vps23ΔUb1, UEV mutation F52AQ117AW125A in pPL2180 This study 
pPL2653 vps23ΔUb2, UEV mutation F52AM107AW125A in pPL2180 This study 
pPL2654 vps23 with UEV mutation F52AL102AW125A in pPL2180 This study 
pPL2655 vps23 with UEV mutation Q117A in pPL2180 This study 
pPL2940 vps23ΔUb1 from pPL2652 in pRS306 This study 
pPL2941 vps23ΔUb2 from pPL2653 in pRS306 This study 
pPL3355 GST-Vps36 GLUE domain (residues 1–289) This study 
pPL3358 GST-Vps36ΔUb GLUE domain (residues 1–289) This study 
pPL3353 VPS36 GLUE domain (residues 1–289) in pET151 This study 
pPL3356 vps36ΔUb (residues 1–289) in pET151 This study 
pPL2251 vps27ΔVps23 in pRS306 PSD1 and PSD2 motifs mutated (AA448ADP and P524AAA) This study 
pPL3230 MVB12 (aa 1–101) in pET151 This study 
pPL3203 MVB12 from S. kluyveri (aa 1–102) in pET151 This study 
pPL3284 MVB12 residues 56–101 in pET151 This study 
pPL3543 MVB12 residues 56–101 F71DQWYK-A71AAAA in pET151 This study 
pPL3542 MVB12 residues 56–101 L92L-D92Q in pET151 This study 
pPL3544 MVB12 residues 56–101 W74Y-R74S in pET151 This study 
pPL3545 MVB12 residues 56–101 K82KPPG-D82DGGT in pET151 This study 
pPL3551 MVB12 residues 56–101 Y79LSKKPP-A79AAAA in pET151 This study 
pPL3552 MVB12 residues 56–101 F71D-S71N in pET151 This study 
pPL3709 mvb12ΔUb1 Y79LSKKPP-A79AAAA with single HA tag in pRS316 This study 
pPL3711 mvb12ΔUb2 K82KPPG-D82DGGT with single HA tag in pRS316 This study 
pPL3713 MVB12 with single HA tag in pRS316 This study 
pPL3780 mvb12ΔUb2 K82KPPG-D82DGGT in pRS306 This study 
pPL3781 mvb12ΔUb1 Y79LSKKPP-A79AAAA in pRS306 This study 
pPL3784 MVB12 in pRS306 This study 
NA MVB12-TAP (TAP-tagged MVB12 driven by PRC1 promoter) Oestreich et al., 2007  
pAO94 mvb12ΔUb1 Y79LSKKPP-A79AAAA -TAP (TAP-tagged mvb12 mutant driven by PRC1 promoter) This study 
NA MVB12-GFP (GFP-tagged MVB12 driven by PRC1 promoter) Oestreich et al., 2007  
pAO104 mvb12ΔUb1 Y79LSKKPP-A79AAAA -GFP (GFP-tagged mvb12 mutant driven by PRC1 promoter) This study 
NA Ub-GST; expression plasmid for Ub fused to GST Katzmann et al., 2001  
pPL1945 GST-Vps27; expression plasmid for residues 353–610 of Vps27 fused to GST Bilodeau et al., 2003  
pGEX3x GST expression plasmid Smith and Johnson, 1988  
NA VPS23-GFP (GFP-tagged Vps23 driven by the endogenous promoter in pRS416) Nickerson et al., 2006  
pPL3982 Vps23ΔUb1-GFP (GFP-tagged Vps23ΔUb1 driven by endogenous promoter) This study 
pPL3979 Vps20-mCherry (mCherry-tagged Vps20 under the control of the endogenous promoter in pRS316) This study 
Plasmid Description Source 
pPL2612 vps36ΔUb (T187F G187A) This study 
pPL2613 VPS36 (−309–1,698 VPS36 and 840 bp of the PHO8 3′UT) in pRS316 This study 
pPL2612 vps36ΔUb (T187F/G187A) in pRS316 This study 
pPL2645 vps36ΔUb (T187F/G187A) as a NotI–Acc65I fragment from pPL2612 in pRS304 This study 
pPL2646 VPS36 as a NotI–Acc65I fragment from pPL2613 in pRS304 This study 
pPL2940 vps23ΔUb1 in pRS306 (F52AQ117AW125A) This study 
pPL2180 VPS23 (−598–1,298) as an XbaI–SacI fragment in pRS316 with BamHI site removed from polylinker with an XbaI–XhoI fill-in ligation This study 
pPL2447 vps23 with UEV mutation F52A in pPL2180 This study 
pPL2448 vps23 with UEV mutation N123A in pPL2180 This study 
pPL2449 vps23 with UEV mutation W125A in pPL2180 This study 
pPL2451 vps23 with UEV mutation S55AD56A in pPL2180 This study 
pPL2452 vps23 with UEV mutation F52AS55AD56A in pPL2180 This study 
pPL2453 vps23 with UEV mutation S55AD56AN123A in pPL2180 This study 
pPL2465 vps23 with UEV mutation S55A in pPL2180 This study 
pPL2466 vps23 with UEV mutation S55AF105A in pPL2180 This study 
pPL2567 vps23 with UEV mutation F105AW125A in pPL2180 This study 
pPL2568 vps23 with UEV mutation F52AF105A in pPL2180 This study 
pPL2569 vps23 with UEV mutation F105A in pPL2180 This study 
pPL2570 vps23 with UEV mutation G57A in pPL2180 This study 
pPL2571 vps23 with UEV mutation G57AF105A in pPL2180 This study 
pPL2578 vps23 with UEV mutation F52AW125A in pPL2180 This study 
pPL2651 vps23 with UEV mutation F52AN123AW125A in pPL2180 This study 
pPL2652 vps23ΔUb1, UEV mutation F52AQ117AW125A in pPL2180 This study 
pPL2653 vps23ΔUb2, UEV mutation F52AM107AW125A in pPL2180 This study 
pPL2654 vps23 with UEV mutation F52AL102AW125A in pPL2180 This study 
pPL2655 vps23 with UEV mutation Q117A in pPL2180 This study 
pPL2940 vps23ΔUb1 from pPL2652 in pRS306 This study 
pPL2941 vps23ΔUb2 from pPL2653 in pRS306 This study 
pPL3355 GST-Vps36 GLUE domain (residues 1–289) This study 
pPL3358 GST-Vps36ΔUb GLUE domain (residues 1–289) This study 
pPL3353 VPS36 GLUE domain (residues 1–289) in pET151 This study 
pPL3356 vps36ΔUb (residues 1–289) in pET151 This study 
pPL2251 vps27ΔVps23 in pRS306 PSD1 and PSD2 motifs mutated (AA448ADP and P524AAA) This study 
pPL3230 MVB12 (aa 1–101) in pET151 This study 
pPL3203 MVB12 from S. kluyveri (aa 1–102) in pET151 This study 
pPL3284 MVB12 residues 56–101 in pET151 This study 
pPL3543 MVB12 residues 56–101 F71DQWYK-A71AAAA in pET151 This study 
pPL3542 MVB12 residues 56–101 L92L-D92Q in pET151 This study 
pPL3544 MVB12 residues 56–101 W74Y-R74S in pET151 This study 
pPL3545 MVB12 residues 56–101 K82KPPG-D82DGGT in pET151 This study 
pPL3551 MVB12 residues 56–101 Y79LSKKPP-A79AAAA in pET151 This study 
pPL3552 MVB12 residues 56–101 F71D-S71N in pET151 This study 
pPL3709 mvb12ΔUb1 Y79LSKKPP-A79AAAA with single HA tag in pRS316 This study 
pPL3711 mvb12ΔUb2 K82KPPG-D82DGGT with single HA tag in pRS316 This study 
pPL3713 MVB12 with single HA tag in pRS316 This study 
pPL3780 mvb12ΔUb2 K82KPPG-D82DGGT in pRS306 This study 
pPL3781 mvb12ΔUb1 Y79LSKKPP-A79AAAA in pRS306 This study 
pPL3784 MVB12 in pRS306 This study 
NA MVB12-TAP (TAP-tagged MVB12 driven by PRC1 promoter) Oestreich et al., 2007  
pAO94 mvb12ΔUb1 Y79LSKKPP-A79AAAA -TAP (TAP-tagged mvb12 mutant driven by PRC1 promoter) This study 
NA MVB12-GFP (GFP-tagged MVB12 driven by PRC1 promoter) Oestreich et al., 2007  
pAO104 mvb12ΔUb1 Y79LSKKPP-A79AAAA -GFP (GFP-tagged mvb12 mutant driven by PRC1 promoter) This study 
NA Ub-GST; expression plasmid for Ub fused to GST Katzmann et al., 2001  
pPL1945 GST-Vps27; expression plasmid for residues 353–610 of Vps27 fused to GST Bilodeau et al., 2003  
pGEX3x GST expression plasmid Smith and Johnson, 1988  
NA VPS23-GFP (GFP-tagged Vps23 driven by the endogenous promoter in pRS416) Nickerson et al., 2006  
pPL3982 Vps23ΔUb1-GFP (GFP-tagged Vps23ΔUb1 driven by endogenous promoter) This study 
pPL3979 Vps20-mCherry (mCherry-tagged Vps20 under the control of the endogenous promoter in pRS316) This study 

NA, not applicable.

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