Table 2.
Yeast strains used in this study
| Strain | Genotype | Used to purify | Source |
|---|---|---|---|
| RPY753 | PGAL1-ZZ-Tev-GFP-3xHA-GST-DYN1331kDa-gs-DHA-KanR; pac1Δ::klURA3; ndl1Δ::cgLEU2 | GST-dynein331 kDa | (Huang et al., 2012) |
| RPY780 | ZZ-Tev-3xHA-DYN1-gs-DHA-KanR nip100Δ pac1Δ::HygroR ndl1Δ::cgLEU2 | Full-length dynein | (Huang et al., 2012) |
| RPY816 | PGAL1-8HIS-ZZ-Tev-PAC1; dyn1D::cgLEU2; ndl1D::HPH | Lis1 | Julie Huang |
| RPY1084 | PGAL1-ZZ-Tev-GFP-3xHA-SNAP-gs-GST-DYN1331kDa-gs-DHA-KanR | Dynein for DNA labeling | (Derr et al., 2012) |
| RPY1099 | PGAL1-8HIS-ZZ-Tev-KIP2-g-FLAG-ga-SNAP−KanR | Kip2 | This work |
| RPY1235 | PGAL1-ZZ-Tev-GFP-3xHA-GST-DYN1E3197K-gs-DHA-KanR | Polarity marker dynein for chassis experiments | (Redwine et al., 2012) |
| RPY1536 | PGAL1-ZZ-Tev-GFP-3xHA-GST-DYN1K3116A, K3117A, E3122A, R3124A-gs-DHA-KanR | Weak dynein | This work |
All strains were made in the W303a background (MATa his3-11,15 ura3-1 leu2-3,112 ade2-1 trp1-1) with genes encoding the proteases Pep4 and Prb1 deleted (prb1Δ; pep4Δ::HIS5). DHA and SNAP refer to the HaloTag (Promega) and SNAP-tag (NEB), respectively. Tev indicates a Tev protease cleavage site. PGAL1 denotes the galactose promoter, which was used to induce protein expression. Amino acid spacers are indicated by g (glycine), ga (glycine–alanine), and gs (glycine–serine).