KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit polyclonal anti-CPY serum | Mousley et al. (2012) | RRID: AB_102312 |
| Bacterial and Virus Strains | ||
| BL21 (DE3) | New England Biolabs | BL21 (DE3) |
| Chemicals, Peptides, and Recombinant Proteins | ||
| His6-Kes1 | Li et al. (2002) | N/A |
| His6-Kes1K109A | This study | N/A |
| His6-Kes1 K109Q | This study | N/A |
| His6-Kes1 K109R | This study | N/A |
| His6-Kes1AcK109 | This study | N/A |
| Critical Commercial Assays | ||
| SensiFast Sybr Lo-Rox Kit | Bioline | N/A |
| VIIA 7 Real-Time PCR System | Applied Biosystems | N/A |
| Q5 High-Fidelity DNA Polymerase kit | New England Biolabs #E0555S | N/A |
| Experimental Models: Organisms/Strains | ||
| S. cerevisiae | ||
| MATa, sec14-1, ura3-52, his3-200, lys2-801 | Bankaitis et al. (1989) | CTY1-1A |
| MATa, sec14-1, pct10, ura3-52, his3-200, lys2-801 | Cleves et al. (1991) | CTY102 |
| MATa, sec14-1, kes10, ura3-52, his3-200, lys2-801 | Fang et al. (1996) | CTY159 |
| MATa, sec14-1, cki1-1, ura3-52, his3-200, lys2-801 | Cleves et al. (1991) | CTY160 |
| MATa, ura3-52, his3-200, lys2-801 | Bankaitis et al. (1989) | CTY182 |
| MATa, sec14-1, ura3-52, his3-200, lys2-801, cki1Δ::URA3 | This study | CTY392 |
| MATa, ura3-52, his3-200, lys2-801, cki1Δ::URA3 | This study | CTY393 |
| MATa, ura3-52, his3-200, lys2-801, pct1Δ::URA3 | This study | CTY1784 |
| MATa, ura3-52, his3-200, lys2-801, kes1Δ::URA3 | This study | CTY1683 |
| MATa, ura3-52, his3-200, lys2-801, cki1Δ::URA3 | This study | CTY1784 |
| CTY1683 leu2Δ::KES1-3XFLAG::HIS3 | This study | JHY2 |
| CTY1683 leu2Δ::kes1 K109A-3XFLAG::HIS3 | This study | JHY3 |
| CTY1683 leu2Δ::kes1 K109Q-3XFLAG::HIS3 | This study | JHY4 |
| CTY1683 leu2Δ::kes1 K109R-3XFLAG::HIS3 | This study | JHY5 |
| CTY159 leu2Δ::KES1-3XFLAG::HIS3 | This study | JHY6 |
| CTY159 leu2Δ::kes1 K109A-3XFLAG::HIS3 | This study | JHY7 |
| CTY159 leu2Δ::kes1 K109Q-3XFLAG::HIS3 | This study | JHY8 |
| CTY159 leu2Δ::kes1 K109R-3XFLAG::HIS3 | This study | JHY9 |
| CTY159 leu2Δ::kes1 K168Q-3XFLAG::HIS3 | This study | JHY10 |
| CTY159 leu2Δ::kes1 K168R-3XFLAG::HIS3 | This study | JHY11 |
| CTY182 osh1Δ::URA3 | This study | JHY14 |
| CTY182 osh2Δ::URA3 | This study | JHY15 |
| CTY182 osh3Δ::URA3 | This study | JHY16 |
| CTY182 osh5Δ::URA3 | This study | JHY17 |
| CTY182 osh6Δ::URA3 | This study | JHY18 |
| CTY182 osh7Δ::URA3 | This study | JHY19 |
| CTY1-1A osh1Δ::URA3 | This study | JHY20 |
| CTY1-1A osh2Δ::URA3 | This study | JHY21 |
| CTY1-1A osh3Δ::URA3 | This study | JHY22 |
| CTY1-1A osh5Δ::URA3 | This study | JHY23 |
| CTY1-1A osh6Δ::URA3 | This study | JHY24 |
| CTY1-1A osh7Δ::URA3 | This study | JHY25 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 DCP2-GFP::HIS3MX6 | Invitrogen | CJM1 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 XRN1-GFP::HIS3MX6 | Invitrogen | CJM2 |
| MATa his3Δ1 leu2Δ0 lys2Δ0 ura3Δ0rim15Δ::KanMX4 | Dharmacon | CJM3 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 | Brachmann et al. (1998) | BY4741 |
| MATa his3Δ1 leu2Δ0 lys2Δ0 ura3Δ0 | Brachmann et al. (1998) | BY4742 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 eaf1 Δ::KANMX | GE Life Sciences | YKB3333 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 rtt109Δ::KANMX | GE Life Sciences | YKB3484 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 elp3Δ::KANMX | GE Life Sciences | YKB3482 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 gcn5Δ::KANMX | GE Life Sciences | YKB3489 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 hat1 Δ::KANMX | GE Life Sciences | YKB3485 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sas2Δ::KANMX | GE Life Sciences | YKB3486 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sas3Δ::KANMX | GE Life Sciences | YKB3487 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 hpa2Δ::KANMX | GE Life Sciences | YKB3488 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 hpa3Δ::KANMX | GE Life Sciences | YKB3297 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sec14ts-NATMX eaf1Δ::KANMX | This study | YKB3935 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sec14ts-NATMX rtt109Δ::HIS3 | This study | YKB4074 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sec14ts-NATMX elp3Δ::KANMX | This study | YKB3444 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sec14ts-NATMX gcn5Δ::KANMX | This study | YKB4080 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sec14ts-NATMX hat1Δ::KANMX | This study | YKB3442 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sec14ts-NATMX sas2Δ::KANMX | This study | YKB3447 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sec14ts-NATMX sas3Δ::KANMX | This study | YKB3449 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sec14ts-NATMX hpa2Δ::KANMX | This study | YKB4065 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sec14ts-NATMX hpa3Δ::KANMX | This study | YKB3451 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 eaf7Δ::KANMX | This study | YKB3292 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 esa1 Δ::HIS3 esa1ts-URA3 | GE Life Sciences | YKB4236 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sec14ts-NATMX eaf7Δ::KANMX | This study | YKB4068 |
| MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 sec14ts-NATMX esa1Δ::HIS3 esa1ts-URA3 | This study | YKB4242 |
| Recombinant DNA | ||
| Plasmids | ||
| YCp(HIS3) | Sikorski and Hieter (1989) | pRS413 |
| pEVOL (mmAcKRs, araC) | Hsu etal. (2016) | pEVOL-MmAcKRS |
| YCp(LEU2) | Sikorski and Hieter (1989) | pRS415 |
| pRS413 Ptet::KES1Y97F | This study | pKB302 |
| pRS415 Ptet::KES1 | This study | pKB321 |
| YCp(URA3, Pdox::KES1) | Mousley et al. (2012) | pCJM1 |
| YCp(URA3, PDOX:: KES1Y97F) | Mousley et al. (2012) | pCJM2 |
| YCp(URA3, PDOX:: kes1K109A) | Mousley et al. (2012) | pCJM3 |
| pRS313 Cki11−200:: Myc6xHIS3 | Ramachandran and Herman (2011) | pPHY2328 |
| pQE30 (KES1) | Li et al. (2002) | pRE540 |
| pQE30 (KES1Y97F) | This study | pRE1454 |
| pQE30 (kes1K109A) | This study | pJH1 |
| pQE30 (kes1K109Q) | This study | pJH2 |
| pQE30 (kes1K109R) | This study | pJH3 |
| pQE30 (kes1K109amber) | This study | pJH4 |
| pQE30 (KES1K168Q) | This study | pJH5 |
| pQE30 (KES1K168R) | This study | pJH6 |
| pDR196(GFP-SNC1, URA3) | This study | pJH7 |
| Software and Algorithms | ||
| Nikon NIS Elements software package version 4.10 | Nikon, Melville, NY | N/A |
| ImageJ version 1.47t | National Institute of Health | N/A |
| Accucomp Beckman Z2 channelyzer software package | Beckman Instruments | N/A |
| R software package “Lattice” | Trellis Graphics (Cleveland, OH) | Lattice graphics |