TABLE 2:
Plasmids used in this study.
| Name | Properties | Source | Figures |
|---|---|---|---|
| pFA6a-2 × YEGFP-kanMX | PCR template for generation of C-terminally 2 × GFP-fused Atg15 | Laboratory stock | |
| pYM-N9 | PCR template for amplification of the ADH1 promoter-YEGFP fragment | ( Janke et al., 2004) | |
| pFA6a-hphNT1 | PCR template for gene deletion | ( Janke et al., 2004) | |
| pFA6a-natNT2 | PCR template for gene deletion | ( Janke et al., 2004) | |
| YGPM21o18 | PCR template for amplification of the PHO8 fragment | Dharmacon | |
| YPGM23k01 | PCR template for amplification of the ATG15 fragment | Dharmacon | |
| pYEX-BX | 2μ plasmid for expression of a protein from the CUP1 promoter | Clontech | |
| pYEX-BX[prApe1] | 2μ plasmid for expression of prApe1 from the CUP1 promoter | ( Suzuki et al., 2013) | 1, S1 |
| pRS314 | Centromeric plasmid (TRP1 marker) | ( Sikorski and Hieter, 1989) | |
| pRS314[mNeonGreen-Atg8] | Centromeric plasmid for expression of mNeonGreen-Atg8 from the ATG8 promoter | ( Hirata et al., 2017) | 1, S1 |
| pRS316 | Centromeric plasmid (URA3 marker) | ( Sikorski and Hieter, 1989) | 5C–D, 6B–E, S3 |
| pRS316[GFP-Atg15TMD] | Centromeric plasmid for expression of YEGFP-Atg15(2–36) from the ADH1 promoter | This study | 4B–C, 5B–D |
| pRS316[GFP-Atg15FL] | Centromeric plasmid for expression of YEGFP-Atg15(2–520) from the ADH1 promoter | This study | 5B–D, S3 |
| pRS316[GFP-Atg15ΔTMD] | Centromeric plasmid for expression of YEGFP-Atg15(37–520) from the ADH1 promoter | This study | 5B–D |
| pRS316[GFP-Pho8TMD-Atg15C] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(37–520) from the ADH1 promoter | This study | S2, 6B–E, 7B–E, 8B–E, 9, 10, 11B–E, S2, S3, S5, S6, S7 |
| pRS316[GFP-Pho8TMD] | Centromeric plasmid for expression of YEGFP-Pho8TMD from the ADH1 promoter | This study | 6B–E, 7B–E, 8B–E, 9, 10, 11B–E, S5, S6, S7 |
| pRS316[GFP-Pho8TMD-Atg15C(S332A)] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(S332A) from the ADH1 promoter | This study | 7B–E |
| pRS316[GFP-Pho8TMD-Atg15C(D387A)] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(D387A) from the ADH1 promoter | This study | 7B–E |
| pRS316[GFP-Pho8TMD-Atg15C(D421A)] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(D421A) from the ADH1 promoter | This study | 7B–E |
| pRS316[GFP-Pho8TMD-Atg15C(H435A)] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(H435A) from the ADH1 promoter | This study | 7B–E |
| pRS316[GFP-Pho8TMD-Atg15ΔC50] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(37–470) from the ADH1 promoter | This study | 8B–E, S5 |
| pRS316[GFP-Pho8TMD-Atg15ΔC54] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(37–466) from the ADH1 promoter | This study | 8B–E, 11B–E, S5, S7 |
| pRS316[GFP-Pho8TMD-Atg15ΔC55] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(37–465) from the ADH1 promoter | This study | 8B–E, S5 |
| pRS316[GFP-Pho8TMD-Atg15C(W466A)] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(W466A) from the ADH1 promoter | This study | 9, 10 |
| pRS316[GFP-Pho8TMD-Atg15ΔN49] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(50–520) from the ADH1 promoter | This study | 11B–E, S6, S7 |
| pRS316[GFP-Pho8TMD-Atg15ΔN49ΔC54] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(50–466) from the ADH1 promoter | This study | 11B–E, S7 |
| pRS423 | 2μ plasmid (HIS3 marker) | ( Sikorski and Hieter, 1989) | |
| pCK3 | pRS423 background plasmid for overexpression of pho8Δ60 | This study | S3 |
| pRS316[GFP-Pho8TMD-Atg15ΔN42] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(43–520) from the ADH1 promoter | This study | S6 |
| pRS316[GFP-Pho8TMD-Atg15ΔN63] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(64–520) from the ADH1 promoter | This study | S6 |
| pRS316[GFP-Pho8TMD-Atg15ΔN70] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(71–520) from the ADH1 promoter | This study | S6 |
| pRS316[GFP-Pho8TMD-Atg15ΔN90] | Centromeric plasmid for expression of YEGFP-Pho8TMD-Atg15(91–520) from the ADH1 promoter | This study | S6 |