Table 1.
Yeast strains used in this study
| Strain | Genotypea | Source |
|---|---|---|
| JY426 | MATa, leu2-3,-112, ura3-52, his4-34, fus1-Δ1, fus2-Δ3 | Cold Spring Harbor Laboratory |
| JY429 | MATα, trp1Δ1, ura3-52, cyh2, fus1-Δ1, fus2-Δ3 | Cold Spring Harbor Laboratory |
| SEY6210 | MATα, leu2-3,-112, ura3-52, his3-Δ200, trp1-Δ901, lys2-801, suc2-Δ9 | S. Emr |
| SEY6211 | MATa, leu2-3,-112, ura3-52, his3-Δ200, trp1-Δ901, ade2, suc2-Δ9 | S. Emr |
| RAY399 | MATα/a, leu2-3,-112/leu2-3,-112, ura3-52/ura3-52, his3-Δ200/his3-Δ200, trp1-Δ901/trp1-Δ901, LYS2/lys2-801, ADE2/ade2, suc2-Δ9/suc2-Δ9 | This studyb |
| RAY485 | RAY399 CDC42/cdc42-Δ1::LEU2 | This study |
| RAY513 | SEY6210 cdc42-Δ1::LEU2 with pRS424GALGFPCDC42 | This studyc |
| RAY563 | SEY6210 sph1-Δ1::HIS3 | Arkowitz and Lowe (1997) |
| RAY567 | SEY6211 sph1-Δ1::HIS3 | Arkowitz and Lowe (1997) |
| RAY950 | MATa, leu2-3,-112, ura3-52, his3-Δ200, trp1-Δ901, lys2-801, ade2, cdc24::LEU2 with pRS416GalHis6CDC24 | Nern and Arkowitz (1998) |
| RAY1042 | RAY950 with pRS414CDC24 instead of pRS416GalHis6CDC24 | Nern and Arkowitz (1998) |
| RAY1052 | SEY6211 cdc24-Δ1::LoxP with pEG(KT)CDC24 | Nern and Arkowitz (2000a) |
| RAY1142 | SEY6211 cdc24::TRP1 cdc24-m1, bud1Δ::LoxP HIS5Sp LoxP | Nern and Arkowitz (1999) |
| RAY1487 | SEY6211 cdc24::TRP1 CDC24, URA3::GFPBUD1 | Nern and Arkowitz (2000a) |
| RAY1556 | SEY6210 cdc42-Δ1::LEU2 with pRS416CDC42 | This studyd |
| RAY1565 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36M, P69T] | This studyd,e |
| RAY1635 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36M] | This studyd,f |
| RAY1638 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[F37I] | This studyd,f |
| RAY1640 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36A, A41G] | This studyd,f |
| RAY1685 | RAY950 with pRS414cdc24-m6 instead of pRS416GalHis6CDC24 | Barale et al. (2004) |
| RAY1728 | SEY6211 cdc24-Δ1::LoxP, cdc42-Δ1::LEU2 with pRS414CDC24 and pRS416CDC42 | This studyg |
| RAY1772 | SEY6211 cdc42-Δ2::HIS5Sp with pRS416CDC42 | This study |
| RAY1793 | SEY6211 cdc24-Δ1::LoxP, cdc42-Δ1::LEU2 with pRS414CDC24 and pRS416cdc42[V36V]i | This studyh |
| RAY1801 | SEY6211 cdc24-Δ1::LoxP, cdc42-Δ1::LEU2 with pRS414cdc24-m6 and pRS416cdc42[V36V]i | This studyj |
| RAY1830 | SEY6211 cdc24-Δ1::LoxP, cdc42-Δ1::LEU2 with pRS414CDC24 and pRS413GFPCDC42 | This studyh |
| RAY1833 | SEY6211 cdc24-Δ1::LoxP, cdc42-Δ1::LEU2 with pRS414cdc24-m6 and pRS413GFPCDC42 | This study |
| RAY1907 | SEY6210 sph1-Δ1::HIS3, URA3::GFPBUD1 | This study |
| RAY1912 | SEY6210 cdc42-Δ1::LEU2 with pRS414cdc42[V36V]i | This study |
| RAY1914 | SEY6210 cdc42-Δ1::LEU2 with pRS414cdc42[V36M] | This study |
| RAY1918 | SEY6211 cdc42-Δ2::HIS5Sp with pRS414cdc42[V36V]i | This study |
| RAY1920 | SEY6211 cdc42-Δ2::HIS5Sp with pRS414cdc42[V36M] | This study |
| RAY1926 | SEY6211 cdc42-Δ2::HIS5Sp with pRS416cdc42[V36M] | This study |
| RAY1951 | SEY6211 cdc24-Δ1::LoxP, cdc42-Δ1::LEU2 with pRS414CDC24 and pRS413GFPcdc42[V36M] | This study |
| RAY1952 | SEY6211 cdc24-Δ1::LoxP, cdc42-Δ1::LEU2 with pRS414CDC24 and pRS416cdc42[V36M] | This studyk |
| DMY13 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36W] | This studyd |
| DMY14 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36C] | This studyd |
| DMY15 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36F] | This studyd |
| DMY16 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36L] | This studyd |
| DMY17 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36I] | This studyd |
| DMY18 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36A] | This studyd |
| DMY19 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36T] | This studyd |
| DMY20 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36Y] | This studyd |
| DMY21 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36Q] | This studyd |
| DMY22 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36S] | This studyd |
| DMY23 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36H] | This studyd |
| DMY25 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36K] | This studyd |
| DMY26 | SEY6210 cdc42-Δ1::LEU2 with pRS416cdc42[V36R] | This studyd |
a HIS5Sp refers to HIS5 from Schizosaccharomyces pombe.
b Constructed by mating SEY6210 and SEY6211.
c Plasmid pRS424GALGFPCDC42 was transformed into RAY399 and tetrads were dissected.
d Indicated plasmid pRS416CDC42 or cdc42 mutant was transformed into RAY513 and pRS424GALGFPCDC42 was cured.
e Isolated in first cdc42 mutant screen.
f Isolated in second cdc42 mutant screen.
g RAY1052 was transformed with pRS414CDC24 and plasmid pEG(KT)CDC24 was cured. This strain was then crossed to RAY1556 followed by sporulation.
h Constructed by transformation of RAY1728 with pRS413GFPCDC42 and pRS416CDC42 was cured (RAY1830). Subsequently strain was transformed with indicated pRS416cdc42 plasmid and plasmid pRS413GFPCDC42 was cured.
i Cdc42[V36V] was generated by site-directed mutagenesis of Cdc42p replacing Val 36 with Val (in addition to a silent SnaBI site).
j Constructed by transformation of RAY1793 with pRS313CDC24 and pRS414CDC24 was cured. Subsequently strain was transformed with pRS414cdc24-m6 plasmid and plasmid pRS313CDC24 was cured.
k Constructed by transformation of RAY1793 with pRS413GFPCDC42 and pRS416cdc42[V36V] was cured. Subsequently the strain was transformed with pRS416cdc42[V36M] plasmid and plasmid pRS413GFPCDC42 was cured.