Table 1.
Integrated alleles of CDC42
| Allelea | Mutation | Growth (YPD)b | Growth (YPD + 3% formamide)c | Restriction sited | DDYe |
|---|---|---|---|---|---|
| CDC42 | None | Wild type (11–37°C) | Wild type (20–37°C) | 1300 | |
| cdc42-1f | Multipleh | Conditional (11–30°C) | Conditional (25°C) | NlaIV (−) | 1302 |
| cdc42-101 | K5A | Conditional (11–30°C) | Lethal | NheI | 1304 |
| cdc42-102 | D11A | Wild type | Conditional (30°C) | BanI | 1306 |
| cdc42-105 | E127A,K128A | Wild type | Wild type | BbvI | 1308 |
| cdc42-107 | R131A,R133A,R135A | Wild type | Wild type | HindIII | 1310 |
| cdc42-108 | R147A,E148A,K150A | Wild type | Wild type | BsrI | 1312 |
| cdc42-109 | E140A | Wild type | Wild type | HhaI | 1314 |
| cdc42-110 | R144A | Wild type | Wild type | NheI | 1316 |
| cdc42-111 | K16A | Lethal | Lethal | HhaI | 1369 |
| cdc42-112 | D31A | Wild type | Wild type | BbvI | 1318 |
| cdc42-113 | D38A | Lethal | Lethal | NruI | 1371 |
| cdc42-114 | D48A,E49A | Wild type | Wild type | BanI | 1320 |
| cdc42-115 | D57A | Lethalg | Lethal | EagI(−) | 1373 |
| cdc42-116 | E62A,D63A | Conditional (20–37°C) | Lethal | BbvI | 1322 |
| cdc42-117 | D65A,R66A,R68A | Conditional (20–37°C)/slow | Lethal | PvuI(−) | 1324 |
| cdc42-118 | D76A | Conditional (11–30°C) | Conditional (25°C) | PstI | 1326 |
| cdc42-119 | E100A | Wild type | Conditional (20–30°C) | PvuII | 1328 |
| cdc42-120 | E91A,K94A,E95A,K96A | Wild type | Conditional (20–34°C) | PstI | 1330 |
| cdc42-121 | D170A,E171A | Wild type | Conditional (20–25°C) | PstI | 1332 |
| cdc42-122 | E178A | Wild type | Wild type | HhaI | 1334 |
| cdc42-123 | R163A,K166A | Conditional (11–30°C) | Lethal | MscI | 1336 |
| cdc42-124 | K183A,K184A,K186A,K187A | Conditional (11–34°C) | Conditional (20–25°C) | BfaI | 1338 |
| cdc42-125 | K153A,E156A | Lethal | Lethal | BsiWI | 1383 |
| cdc42-126 | Y32K | Conditional (25°C)/slow | Lethal | BanI | 1340 |
| cdc42-127 | V33A | Wild type | Wild type | HhaI | 1342 |
| cdc42-128 | T35A | Lethal | Lethal | PstI | 1386 |
| cdc42-129 | V36T | Conditional (20–34°C) | Lethal | Psp1406I | 1344 |
| cdc42-130 | F37Y | Wild type | Wild type | AccI | 1346 |
| cdc42-131 | N39A | Wild type | Lethal | SfaNI | 1348 |
| cdc42-132 | Y40K | Lethal | Lethal | AlwNI | 1390 |
| cdc42-133 | Y40C | Lethal | Lethal | HhaI | 1391 |
| cdc42-134 | D118A | Lethal | Lethal | Sau3AI(−) | 1392 |
| cdc42-135 | R120A,D121A,D122A,K123A | Lethal | Lethal | SacII | 1393 |
| cdc42-136 | R120A | Wild type | Conditional (20–34°C) | BglII | 1350 |
| cdc42-137 | D121A | Wild type | Wild type | BsrBI | 1352 |
| cdc42-138 | D122A | Wild type | Wild type | SfaNI | 1354 |
| cdc42-139 | K123A | Wild type | Wild type | BsaHI | 1356 |
| cdc42-140 | H102A,H103A,H104A | Wild type | Conditional (20–34°C) | PstI | 1358 |
a Each allele, except cdc42-1, is linked to LEU2+, as shown in Figure 1C.
b Growth phenotype of haploid cells on YPD plates at 11, 14, 20, 25, 30, 34, and 37°C. The permissive temperature range is indicated in parentheses. Relevant genotype is cdc42∶LEU2+ ura3-52 leu2-3,112 his3Δ200 lys2-801am.
c Growth phenotype of haploid cells on YPD plates containing 3% (v/v) formamide.
d Diagnostic restriction site added during site-directed mutagenesis to mark mutation. (−) denotes loss of the restriction site.
e The DDY strain number is listed for each allele of CDC42 in a MATa ura3-52 leu2-3,112 his3Δ200 lys2-801am strain. The background of each strain is S288C. Isogenic MATα haploids are designated by the next ascending odd strain number (not shown). For recessive lethal cdc42 alleles, the listed strain number is for a diploid strain with the relevant genotype MATa/MATα, cdc42/CDC42, ade2-1/ADE2, ura3-52/ura3-52, leu2-3,112/leu2-3,112, his3Δ200/his3Δ200, lys2-801am/LYS2.
f Allele isolated by Adams et al. (1990). Strain congenic with all other strains in this study, except LEU2+ is at LEU2 and unlinked to cdc42-1.
g Mutation may be dominant lethal. Only one heterozygote recovered.
h See Miller and Johnson (1997).