Table 2.
Phosphorylation of Ser 197 but not Ser 496 is essential for Kar9p function
| Cross | Viability | Tetrads (PD:TT:NPD) | Double mutants (viable/predicted) |
|---|---|---|---|
| kar9-AA × dyn1Δ | Not viable | 19 (5:10:4) | 5/18 |
| S197A × dyn1Δ | Not viable | 19 (7:10:2) | 1/14 |
| S496A × dyn1Δ | Viable | 18 (1:15:2) | 19/19 |
| S197A × jnm1Δ | Not viable | 20 (2:13:5) | 1/23 |
| S496A × jnm1Δ | Viable | 19 (2:16:1) | 17/18 |
| S197A × bik1Δ | Not viable | 14 (3:9:2) | 0/13 |
| S496A × bik1Δ | Viable | 15 (3:10:2) | 13/14 |
| S197A × bim1Δ | Viable | 17 (3:11:3) | 15/17 |
| S496A × bim1Δ | Viable | 20 (2:14:4) | 19/22 |
| S197A ×clb4Δ | Viable | 15 (3:9:3) | 14/15 |
| S496A ×clb4Δ | Viable | 15 (2:10:2) | 13/14 |
| kar9-AA ×clb4Δ | Viable | 18 (2:11:5) | 20/21 |
| kar9Δ ×clb4Δ | Viable | 19 (5:12:2) | 16/16 |
| dyn1Δ ×clb4Δ | Sick | 18 (3:10:5) | 11/20 |
| jnm1Δ ×clb4Δ | Sick | 19 (1:15:3) | 3/21 |
Double mutants were generated by meiotic crosses and scored for growth after 2-3 d. Progeny exhibiting no growth or forming microcolonies ∼10-fold smaller than single mutant or wild-type cells (see Supplemental Figure S1) were scored as “inviable.” Microcolonies also exhibited poor growth upon streaking to new media. The dyn1Δclb4Δ and jnm1Δclb4Δ double mutants were scored as “sick” as these colonies were smaller than those produced by single mutants but did not exhibit phenotypes as severe as those observed for other double mutants (see Supplemental Figure S1). Of the 20 predicted dyn1Δ × clb4Δ double mutants, 11 produced colonies equivalent to the size of those formed by single mutants. Six produced small colonies that displayed severely retarded growth, and three exhibited no growth at all. For the 21 predicted jnm1Δ × clb4Δ double mutants, three formed colonies similar to the size of single mutants. Eight produced small colonies that exhibited severely retarded growth, and 10 exhibited no growth. The following parental strains were used in these crosses: Kar9p-AA-tap (yRM6170), Kar9p-S197A-tap (yRM6168), Kar9p-S496A-tap (yRM6015), dyn1Δ (yRM1094), jnm1Δ (yRM469), bik1Δ (yRM565), bim1Δ (yRM2060), clb4Δ (yRM6152), and kar9Δ (yRM369). The clb4Δ strain yRM6026 was used for crosses with dyn1Δ, jnm1Δ, and kar9Δ.