Table 1.
Plasmids used in this study
| Plasmid name | Genotype |
|---|---|
| YCpT-MID1-FL | LEU2 CEN4 ARS1 TDH3p:MID1–5xFLAG ampr |
| YCpS-MID1 | LEU2 CEN4 ARS1 MID1p:MID1 ampr |
| YCpS-MID1ΔN23a | LEU2 CEN4 ARS1 MID1p:MID1 (Δ2–23) ampr |
| YCpS-MID1ΔN23-HAa | LEU2 CEN4 ARS1 MID1p:MID1 (Δ2–23)-HAx4 ampr |
| YCpT-MID1ΔN23a | LEU2 CEN4 ARS1 TDH3p:MID1 (Δ2–23) ampr |
| YCpS-MID1Δ210–219 | LEU2 CEN4 ARS1 MID1p:MID1 (Δ210–219) ampr |
| YCpT-MID1Δ210–219 | LEU2 CEN4 ARS1 TDH3p:MID1 (Δ209–219) ampr |
| YCpS-MID1-FL52 | LEU2 CEN4 ARS1 MID1p:MID1-5xFLAG52ampr |
| YCpS-MID1ΔN23-FL52 | LEU2 CEN4 ARS1 MID1p:MID1 (Δ2–23)-5xFLAG52ampr |
| YCpT-MID1(11Q) | LEU2 CEN4 ARS1 TDH3p:MID1 (11Q)bampr |
| YCpS-SUC2C-MID1-FL | LEU2 CEN4 ARS1 MID1p:SUC2C-MID1–5xFLAG ampr |
| YCpS-SUC2CQ-MID1-FL | LEU2 CEN4 ARS1 MID1p:SUC2CQ-MID1–5xFLAG ampr |
| YCpS-MID1(9Q)-FL-SUC2C | LEU2 CEN4 ARS1 MID1p:MID1 (9Q)c-5xFLAG-SUC2C ampr |
| YCpS-MID1(9Q)-FL-SUC2CQ | LEU2 CEN4 ARS1 MID1p:MID1 (9Q)c-5xFLAG-SUC2CQ ampr |
| YCpT-MID1-HIS4C | URA3 CEN4 ARS1 TDH3p:MID1-HIS4C ampr |
| pBCT-CCH1-HIS4C | URA3 CEN4 ARS1 TDH3p:CCH1-HIS4C ampr |
| pCS4-14d | URA3 2 μm-ori HMG1-SUC2-HIS4C ampr |
a Plasmids that express a series of N-terminally truncated forms of the Mid1 and Mid1-HA proteins, including Mid1ΔN32, Mid1ΔN51, Mid1ΔN68, Mid1ΔN112, Mid1ΔN209, and Mid1ΔN219 and their corresponding C-terminally HA-tagged derivatives were also constructed. However, to avoid redundancy, only the three listed plasmids were described as representatives.
b 11Q, a series of MID1 mutants with 11 Asn → Gln substitutions: N32Q, N70Q, N228Q, N238Q, N265Q, N285Q, N291Q, N324Q, N389Q, N407Q, and N536Q.
c 9Q, MID1 mutants with nine Asn → Gln substitutions: N32Q, N70Q, N228Q, N238Q, N275Q, N324Q, N389Q, N407Q, and N536Q.
d A gift from Sengstag (35).