TABLE 1.
S. cerevisiae strains used in this study
| Strain | Relevant genotypea | Source or reference |
|---|---|---|
| YEF473 | MATa/α his3-Δ200/his3-Δ200 leu2-Δ1/leu2-Δ1 lys2-801/lys2-801 trp1-Δ63/trp1-Δ63 ura3-52/ura3-52 | 7 |
| YEF473A | MATahis3-Δ200 leu2-Δ1 lys2-801 trp1-Δ63 ura3-52 | 56 |
| YEF473B | MATα his3-Δ200 leu2-Δ1 lys2-801 trp1-Δ63 ura3-52 | 56 |
| LSY365 | MATarsr1Δ::HIS3 | J. Pringle |
| LSY388 | MATa/α rsr1Δ::HIS3/rsr1Δ::HIS3 | 66 |
| HH615 | MATa/α bud9-Δ1::HIS3/bud9-Δ1::HIS3 | 34 |
| M-2772 | MATα cla4Δ::HIS3 [YCp111/CDC3-GFP] | Transformant of YEF1343 11 |
| M-3491 | MATa [YCp111/CDC12-YFP] | Transformant of YEF473A |
| M-3492 | MATa [YCp111/cdc12G44V, K47E, T48N-YFP] | Transformant of YEF473A |
| M-376 | MATa/α cdc12Δ::TRP1/CDC12 | This studyb |
| M-377 | MATacdc12Δ::TRP1 [YEp24(CDC12)N] | This studyc |
| YNT107 | MATacdc12Δ::TRP1 [YCp111/CDC12] | This studyd |
| M-2863 | MATacdc12Δ::TRP1 [YCp111/CDC12] and [pRS316/GFP-CDC3] | Transformant of YNT107 |
| YNT233 | MATacdc12Δ::TRP1 [YCp111/cdc12G44V] | This studyd |
| M-2864 | MATacdc12Δ::TRP1 [YCp111/cdc12G44V] and [pRS316/GFP-CDC3] | Transformant of YNT233 |
| YNT109 | MATacdc12Δ::TRP1 [YCp111/cdc12K47T] | This studyd |
| M-2865 | MATacdc12Δ::TRP1 [YCp111/cdc12K47T] and [pRS316/GFP-CDC3] | Transformant of YNT109 |
| YNT131 | MATacdc12Δ::TRP1 [YCp111/cdc12G44V, K47E, T48N] | This studyd |
| M-2866 | MATacdc12Δ::TRP1 [YCp111/cdc12G44V, K47E, T48N] and [pRS316/GFP-CDC3] | Transformant of YNT131 |
| M-2840 | MATacdc12Δ::TRP1 [YCp111/cdc12D98A, G101A] | This studyd |
| M-2867 | MATacdc12Δ::TRP1 [YCp111/cdc12D98A, G101A] and [pRS316/GFP-CDC3] | Transformant of M-2840 |
| M-1726 | MATacdc12-6 | This studye |
| M-2962 | MATacdc12-6 [pRS316/GFP-CDC3] | Transformant of M-1726 |
| M-3489 | MATa [YCp111/CDC10-YFP] | Transformant of YEF473A |
| M-3490 | MATa [YCp111/cdc10K45T-YFP] | Transformant of YEF473A |
| DD185 | MATa/α cdc10Δ::HIS3/CDC10 | 15 |
| M-611 | MATacdc10Δ::HIS3 [YEp24/CDC10] | This studyf |
| YNT89 | MATacdc10Δ::HIS3 [YCp111] | This studyg |
| M-2894 | MATacdc10Δ::HIS3 [YCp111] and [pRS316/GFP-CDC3] | Transformant of YNT89 |
| YNT91 | MATacdc10Δ::HIS3 [YCp111/CDC10] | This studyg |
| M-2855 | MATacdc10Δ::HIS3 [YCp111/CDC10] and [pRS316/GFP-CDC3] | Transformant of YNT91 |
| YNT225 | MATacdc10Δ::HIS3 [YCp111/cdc10G42V] | This studyg |
| M-2856 | MATacdc10Δ::HIS3 [YCp111/cdc10G42V] and [pRS316/GFP-CDC3] | Transformant of YNT225 |
| YNT93 | MATacdc10Δ::HIS3 [YCp111/cdc10K45T] | This studyg |
| M-2857 | MATacdc10Δ::HIS3 [YCp111/cdc10K45T] and [pRS316/GFP-CDC3] | Transformant of YNT93 |
| M-3487 | MATa [YCp111/CDC3-YFP] | Transformant of YEF473A |
| M-3488 | MATa [YCp111/cdc3G129V, K132E, T133N-YFP] | Transformant of YEF473A |
| M-379 | MATa/α cdc3Δ::TRP1/CDC3 | This studyb |
| M-381 | MATacdc3Δ::TRP1 [pRSCDC3] | This studyh |
| M-438 | MATacdc3Δ::TRP1 [YCp111/CDC3] | This studyi |
| M-2922 | MATacdc3Δ::TRP1 [YCp111/CDC3] and [pMS76] | Transformant of M-381 |
| M-2914 | MATacdc3Δ::TRP1 [YCp111/cdc3G129V] | This studyi |
| M-2923 | MATacdc3Δ::TRP1 [YCp111/cdc3G129V] and [pMS76] | Transformant of M-2914 |
| M-2915 | MATacdc3Δ::TRP1 [YCp111/cdc3K132T] | This studyi |
| M-2924 | MATacdc3Δ::TRP1 [YCp111/cdc3K132T] and [pMS76] | Transformant of M-2915 |
| M-2916 | MATacdc3Δ::TRP1 [YCp111/cdc3K132E] | This studyi |
| M-2925 | MATacdc3Δ::TRP1 [YCp111/cdc3K132E] and [pMS76] | Transformant of M-2916 |
| M-2917 | MATacdc3Δ::TRP1 [YCp111/cdc3T133N] | This studyi |
| M-2926 | MATacdc3Δ::TRP1 [YCp111/cdc3T133N] and [pMS76] | Transformant of M-2917 |
| M-2918 | MATacdc3Δ::TRP1 [YCp111/cdc3G129V, K132E, T133N] | This studyi |
| M-2927 | MATacdc3Δ::TRP1 [YCp111/cdc3G129V, K132E, T133N] and [pMS76] | Transformant of M-2918 |
| M-3474 | MATa/α cdc3Δ::TRP1/cdc3Δ::TRP1 [YCp111/cdc3G129V, K132E, T133N] | This studyj |
| M-3344 | MATa/α [YCp111/CDC11-YFP] | Transformant of YEF473 |
| M-3352 | MATa/α [YCp111/cdc11K172A, D174A-YFP] | Transformant of YEF473 |
| M-2036 | MATa/α cdc11Δ::TRP1/CDC11 | This studyb |
| M-2283 | MATa/α cdc11Δ::TRP1/CDC11 ura3:URA3:CDC11/ura3 | Transformant of M-2036 |
| M-2332 | MATacdc11Δ::TRP1 ura3:URA3:CDC11 | Segregant from M-2283 |
| M-2333 | MATα cdc11Δ::TRP1 ura3:URA3:CDC11 | Segregant from M-2283 |
| M-2334 | MATa/α cdc11Δ::TRP1/cdc11Δ::TRP1 ura3:URA3:CDC11/ura3:URA3:CDC11 | M-2332 × M-2333 |
| M-2437 | MATacdc11Δ::TRP1 ura3:URA3:CDC11 [YCp111/CDC3-GFP] | Transformant of M-2332 |
| M-2297 | MATacdc11G32E | This studyk |
| M-2298 | MATα cdc11G32E | This studyk |
| M-2344 | MATa/α cdc11G32E/cdc11G32E | M-2297 × M-2298 |
| M-2452 | MATacdc11G32E [YCp111/CDC3-GFP] | Transformant of M-2297 |
| M-3172 | MATacdc11G32E [pMS76] | Transformant of M-2297 |
| M-3174 | MATacdc11G32E cdc3Δ::TRP1 [YCp111/cdc3K132T] and [pMS76] | This studyl |
| M-2042 | MATa/α cdc11Δ::TRP1/CDC11 ura3:URA3:cdc11G32V/ura3 | Transformant of M-2036 |
| M-2323 | MATacdc11Δ::TRP ura3:URA3:cdc11G32V | Segregant from M-2042 |
| M-2324 | MATα cdc11Δ::TRP ura3:URA3:cdc11G32V | Segregant from M-2042 |
| M-2325 | MATa/α cdc11Δ::TRP1/cdc11Δ::TRP1 ura3:URA3:cdc11G32V/ura3:URA3:cdc11G32V | M-2323 × M-2324 |
| M-2440 | MATacdc11Δ::TRP ura3:URA3:cdc11G32V [YCp111/CDC3-GFP] | Transformant of M-2323 |
| M-2723 | MATacdc11Δ::TRP ura3:URA3:cdc11G32V cla4Δ::HIS3 [YCp111/CDC3-GFP] | This studym |
| M-2275 | MATa/α cdc11Δ::TRP1/CDC11 ura3:URA3:cdc11R35T/ura3 | Transformant of M-2036 |
| M-2311 | MATacdc11Δ::TRP1 ura3:URA3:cdc11R35T | Segregant from M-2275 |
| M-2312 | MATα cdc11Δ::TRP1 ura3:URA3:cdc11R35T | Segregant from M-2275 |
| M-2313 | MATa/α cdc11Δ::TRP1/cdc11Δ::TRP1 ura3:URA3:cdc11R35T/ura3:URA3:cdc11R35T | M-2311 × M-2312 |
| M-2443 | MATacdc11Δ::TRP1 ura3:URA3:cdc11R35T [YCp111/CDC3-GFP] | Transformant of M-2311 |
| M-2726 | MATacdc11Δ::TRP ura3:URA3:cdc11R35T cla4Δ::HIS3 [YCp111/CDC3-GFP] | This studyn |
| M-2273 | MATa/α cdc11Δ::TRP1/CDC11 ura3:URA3:cdc11R35E/ura3 | Transformant of M-2036 |
| M-2299 | MATacdc11Δ::TRP1 ura3:URA3:cdc11R35E | Segregant from M-2273 |
| M-2300 | MATα cdc11Δ::TRP1 ura3:URA3:cdc11R35E | Segregant from M-2273 |
| M-2301 | MATa/α cdc11Δ::TRP1/cdc11Δ::TRP1 ura3:URA3:cdc11R35E/ura3:URA3:cdc11R35E | M-2299 × M-2300 |
| M-2446 | MATacdc11Δ::TRP1 ura3:URA3:cdc11R35E [YCp111/CDC3-GFP] | Transformant of M-2299 |
| M-2776 | MATacdc11Δ::TRP1 ura3:URA3:cdc11R35E cla4Δ::HIS3 [YCp111/CDC3-GFP] | This studyo |
| M-2274 | MATa/α cdc11Δ::TRP1/CDC11 ura3:URA3:cdc11S36N/ura3 | Transformant of M-2036 |
| M-2305 | MATacdc11Δ::TRP1/CDC11 ura3:URA3:cdc11S36N | Segregant from M-2274 |
| M-2306 | MATα cdc11Δ::TRP1/CDC11 ura3:URA3:cdc11S36N | Segregant from M-2274 |
| M-2307 | MATa/α cdc11Δ::TRP1/cdc11Δ::TRP1 ura3:URA3:cdc11S36N/ura3:URA3:cdc11S36N | M-2305 × M-2306 |
| M-2449 | MATacdc11Δ::TRP1/CDC11 ura3:URA3:cdc11S36N [YCp111/CDC3-GFP] | Transformant of M-2305 |
| M-367 | MATacdc11Δ::TRP1 [pRSCDC11] | This studyp |
| M-368 | MATα cdc11Δ::TRP1 [pRSCDC11] | This studyp |
| M-432 | MATa/α cdc11Δ::TRP/cdc11Δ::TRP1 [pRSCDC11] | M-367 × M-368 |
| YNT95 | MATacdc11Δ::TRP1 [YCplac111] | This studyq |
| M-2897 | MATacdc11Δ::TRP1 [YCplac111] and [pRS316/GFP-CDC3] | Transformant of YNT95 |
| YNT97 | MATacdc11Δ::TRP1 [YCp111/CDC11] | This studyq |
| M-3367 | MATacdc11Δ::TRP1 [YCp111/CDC11-YFP] | This studyq |
| M-2365 | MATacdc11Δ::TRP1 [YCp111/cdc11G29D] | This studyq |
| M-2859 | MATacdc11Δ::TRP1 [YCp111/cdc11G29D] and [pRS316/GFP-CDC3] | Transformant of M-2365 |
| YNT195 | MATacdc11Δ::TRP1 [YCp111/cdc11G32V, R35E, S36N] | This studyq |
| M-2860 | MATacdc11Δ::TRP1 [YCp111/cdc11G32V, R35E, S36N] and [pRS316/GFP-CDC3] | Transformant of YNT195 |
| M-3214 | MATacdc11Δ::TRP1 [YCp111/cdc11D89A, G92A] | This studyq |
| M-3277 | MATacdc11Δ::TRP1 CDC3:URA3:CDC3-GFP [YCp111/cdc11D89A, G92A] | Transformant of M-3214r |
| M-3186 | MATa/α cdc11Δ::TRP/cdc11Δ::TRP1 [YCp111/cdc11D89A, G92A] | This studys |
| M-3258 | MATacdc11Δ::TRP1 [YCp111/cdc11K172A, D174A] | This studyq |
| M-3374 | MATacdc11Δ::TRP1 [YCp111/cdc11K172, D174A-YFP] | This studyq |
| M-3280 | MATacdc11Δ::TRP1 CDC3:URA3:CDC3-GFP [YCp111/cdc11K172A, D174A] | Transformant of M-3258r |
| M-3472 | MATacdc11Δ::TRP1 CDC3:URA3:CDC3-GFP cla4Δ::HIS3 [YCp111/cdc11K172A, D174A] | This studys |
| M-3473 | MATa/α cdc11Δ::TRP/cdc11Δ::TRP1 [YCp111/cdc11G32V, R35E, S36N] | This studyt |
| M-2618 | MATα cdc11G32E-3HA | Transformant of M-2298u |
| M-2630 | MATa/α cdc11G32E-3HA/cdc11G32E | M-2618 × M-2297 |
| M-2634 | MATa/α cdc11G32E-3HA/CDC11 | M-2618 × YEF473A |
| M-2159 | MATα CDC11-3HA | Transformant of YEF473Bu |
| M-2633 | MATa/α CDC11-3HA/CDC11 | M-2159 × YEF473A |
| EGY48R | MATahis3 trp1 ura3-52 [pSH18-34] | 78v |
Except for strain EGY48R, all strains are congenic with YEF473. Plasmids are indicated by square brackets.
CDC3, CDC11, and CDC12 were deleted in strain YEF473, as described previously (22).
Segregant from M-376 transformed with YEp24(CDC12)N.
Strain M-377 was transformed with the indicated LEU2 plasmid and plated on medium lacking leucine and containing 0.1% 5-fluoroorotic acid to select for loss of YEp24(CDC12)N.
Derived from backcrossing the cdc12-6 strain M-238 (56) 10 times into the YEF473 strain background.
Segregant from DD185 transformed with YEp24/CDC10.
Strain M-611 was transformed with the indicated LEU2 plasmid and plated on medium lacking leucine and containing 0.1% 5-fluoroorotic acid to select for loss of YEp24/CDC10.
Segregant from M-379 transformed with pRSCDC3.
Strain M-381 was transformed with the indicated LEU2 plasmid and plated on medium lacking leucine and containing 0.1% 5-fluoroorotic acid to select for loss of pRSCDC3.
Derived from mating segregants from M-379 transformed with pRSCDC3, followed by transformation with YCp111/cdc3G129V, K132E, T133N and plating on medium lacking leucine and containing 0.1% 5-fluoroorotic acid to select for loss of pRSCDC3.
Derived from backcrossing the cdc11-6 strain JPT194 (1) 10 times into the YEF473 strain background.
Segregant from cross of M-2298 and M-2925.
Segregant from cross of M-2440 and M-2772.
Segregant from cross of M-2443 and M-2772.
Segregant from cross of M-2446 and M-2772.
Segregant from M-2036 transformed with pRSCDC11.
Strain M-367 was transformed with the indicated LEU2 plasmid and plated on medium lacking leucine and containing 0.1% 5-fluoroorotic acid to select for loss of pRSCDC11.
Transformed with ApaI-digested YIp211/CDC3-GFP.
Segregant from cross of M-3280 and YEF1373.
Strain M-432 was transformed with YCp111/cdc11K172A, D174A and plated on medium lacking leucine and containing 0.1% 5-fluoroorotic acid to select for loss of pRSCDC11.
Transformed with the PCR product of primers ML298 and ML299 using as template pFA6a-3HA-His3MX6 (57).
pSH18-34 is a 2μm URA3 plasmid with an 8lexOp-lacZ reporter gene (3).