4. Materials
4.2 Strains and Plasmids
4.2.2 S. cerevisiae strains
The following table shows the S. cerevisiae strains used in this study. A PCR-based method for gene fusions and gene deletions was used (Janke et al., 2004). If no indicated, all used yeast strains derived from the genetic background of SK1. The genomic modifications in the yeast strains were checked by PCR. The mating type was tested either by SPO medium or by cross-linking with the mating type tester strains YR312 and YR320.
Table 6: Used S. cerevisiae strains
Strain Genotype Reference
ESM356 MATa
ura3-53 leu2Δ1 his3Δ200 trp1Δ63
C. Taxis collection ESM357 MATα
ura3-53 leu2∆1 his3∆200 C. Taxis
collection
SGY37 MATa
ura3-52::URA3-lexA-op-LacZ trp1 his3 leu2
(Geissler et al., 1996)
YAA129 MATα
lys2 leu2 ho::LYS2 ura3
his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3 trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1
C. Taxis collection YAA130 MATa
lys2 leu2 ho::LYS2 ura3
his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3 trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1
C. Taxis collection YAA146 MATa /MATα
lys2/lys2 leu2/leu2 ho::LYS2/ho::LYS2 ura3/ura3 his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3/
his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3 trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1/
trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1
C. Taxis collection
YAM281 MATa/MATα
YKS32 MPC70-yeGFP::kanMX/MPC70-yeGFP::kanMX
(Taxis et al., 2005) YCR77 MATa/MATα
ho::LYS2 his3 leu2 trp1 ura3/ho::LYS2 his3 leu2 trp1 ura3
C. Taxis collection YCR329 MATa/MATα
ura3-52 lys2 ho::LYS2/leu2delta ura3-52 lys2 his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3/
his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3 trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1/
trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1
C. Taxis collection
86 YCR484 MATa/MATα
ura3Δ /ura3Δ his3Δ/his3Δ leu2Δ /leu2Δ trp1 Δ/trp1Δ ho::hisG/ho::hisG
MPC54/MPC54-6HA::spHIS5
C. Taxis collection YCR485 MATa/MATα
ura3Δ /ura3Δ his3Δ/his3Δ leu2Δ /leu2Δ trp1 Δ/trp1Δ ho::hisG/ho::hisG
MPC70/MPC70-6HA::spHIS5
C. Taxis collection YCR561 MATa/MATα
ura3Δ /ura3Δ his3Δ/his3Δ leu2Δ /leu2Δ lys2Δ /lys2Δ trp1 Δ/trp1Δ ho::LYS2/ho::LYS2
SPO74/SPO74-9myc::hphNT1
C. Taxis collection YCT730 MATa/MATα
YKS32 MPC54-yeGFP::kanMX/MPC54-yeGFP::kanMX
(Taxis et al., 2005) YDS152 MATa/MATα
ura3Δ /ura3Δ his3Δ/his3Δ leu2Δ /leu2Δ lys2Δ /lys2Δ trp1 Δ/trp1Δ ho::LYS2/ho::LYS2
ADY1/ADY1-9myc::hphNT1
C. Taxis collection YKS32 MATa/MATα
lys2/lys2 ura3/ura3 leu2/LEU2 ho::hisG/ho::LYS (Knop and Strasser, 2000) YMK725 MATa/MATα
lys2/lys2 ura3/ura3 leu2/LEU2 ho::his6/ho::LYS2 spo74Δ hphNT1:: spo74Δ hphNT1
C. Taxis collection
YMJ36 YKS32 MATa (Jungbluth et
al., 2012)
YMJ37 YKS32 MATα (Jungbluth et
al., 2012) YMJ68 MATa/MATα
ady1Δ::natNT2/ady1Δ::natNT2
(Jungbluth et al., 2012) YMJ69 MATa/MATα
ady2Δ::hphNT1/ady2Δ::hphNT1
(Jungbluth et al., 2012) YMJ143 MATα
cyr1Δ::hphNT1
pMJ15 [CYR1 URA3 ARS/CEN]
C. Taxis collection YMJ144 MATa
cyr1Δ::hphNT1
pMJ15 [CYR1 URA3 ARS/CEN]
C. Taxis collection YMJ145 MATα
cyr1Δ::hphNT1
pDS66 [cyr1K1712A URA3 ARS/CEN]
C. Taxis collection YMJ146 MATa
cyr1Δ::hphNT1
pDS66 [cyr1K1712A URA3 ARS/CEN]
C. Taxis collection
YMM1 MATα
MPC70-yemGFP::KanMX cyr1Δ::hphNT1 pMJ15 [CYR1 URA3 ARS/CEN]
Created by transformation of PCR product of primers MPC70-S2 and MPC70-S3 with pYM12 Monomeric as template into YMJ143.
Checked by test PCR with hiskantag2 and MPC70-tag.
This work
YMM2 MATa This work
87 MPC70-yemGFP::KanMX cyr1Δ::hphNT1
pMJ15 [CYR1 URA3 ARS/CEN]
Created by transformation of PCR product of primers MPC70-S2 and MPC70-S3 with pYM12 Monomeric as template into YMJ144.
Checked by test PCR with hiskantag2 and MPC70-tag.
YMM3 MATα
SPO74-yemGFP::KanMX cyr1Δ::hphNT1 pMJ15 [CYR1 URA3 ARS/CEN]
Created by transformation of PCR product of primers SPO74-S2 and SPO74-S3 with pYM12 Monomeric as template into YMJ143.
Checked by test PCR with hiskantag2 and SPO74-tag.
This work
YMM4 MATa
SPO74-yemGFP::KanMX cyr1Δ::hphNT1 pMJ15 [CYR1 URA3 ARS/CEN]
Created by transformation of PCR product of primers SPO74-S2 and SPO74-S3 with pYM12 Monomeric as template into YMJ144.
Checked by test PCR with hiskantag2 and SPO74-tag.
This work
YMM5 MATa/MATα
MPC70-yemGFP::KanMX cyr1Δ::hphNT1/ MPC70-yemGFP::KanMX cyr1Δ::hphNT1
pMJ15 [CYR1 URA3 ARS/CEN]
Created by crossing YMM1 and YMM2.
This work
YMM6 MATa/MATα
SPO74-yemGFP::KanMX cyr1Δ::hphNT1/ SPO74-yemGFP::KanMX cyr1Δ::hphNT1
pMJ15 [CYR1 URA3 ARS/CEN]
Created by crossing YMM3 and YMM4.
This work
YMM7 MATα
MPC54-yemGFP::KanMX cyr1Δ::hphNT1 pMJ15 [CYR1 URA3 ARS/CEN]
Created by transformation of PCR product of primers MPC54-S2 and MPC54-S3 with pYM12 Monomeric as template into YMJ143.
Checked by test PCR with hiskantag2 and MPC54-tag.
This work
YMM8 MATa
MPC54-yemGFP::KanMX cyr1Δ::hphNT1 pMJ15 [CYR1 URA3 ARS/CEN]
Created by transformation of PCR product of primers MPC54-S2 and MPC54-S3 with pYM12 Monomeric as template into YMJ144.
Checked by test PCR with hiskantag2 and MPC54-tag.
This work
YMM9 MATα
MPC54-yemGFP::KanMX cyr1Δ::hphNT1 pDS66 [cyr1K1712A URA3 ARS/CEN]
Created by transformation of PCR product of primers MPC54-S2 and MPC54-S3 with pYM12 Monomeric as template into YMJ145.
Checked by test PCR with hiskantag2 and MPC54-tag.
This work
YMM10 MATα This work
88 ADY1-yemGFP::KanMX cyr1Δ::hphNT1
pMJ15 [CYR1 URA3 ARS/CEN]
Created by transformation of PCR product of primers ADY1-S2 and ADY1-S3 with pYM12 Monomeric as template into YMJ143.
Checked by test PCR with hiskantag2 and ADY1-tag.
YMM11 MATa
ADY1-yemGFP::KanMX cyr1Δ::hphNT1 pMJ15 [CYR1 URA3 ARS/CEN]
Created by transformation of PCR product of primers ADY1-S2 and ADY1-S3 with pYM12 Monomeric as template into YMJ144.
Checked by test PCR with hiskantag2 and ADY1-tag.
This work
YMM12 MATa
ADY1-yemGFP::KanMX cyr1Δ::hphNT1 pDS66 [cyr1K1712A URA3 ARS/CEN]
Created by transformation of PCR product of primers ADY1-S2 and ADY1-S3 with pYM12 Monomeric as template into YMJ146.
Checked by test PCR with hiskantag2 and ADY1-tag.
This work
YMM13 MATα
MPC70-yemGFP::KanMX cyr1Δ::hphNT1 pDS66 [cyr1K1712A URA3 ARS/CEN]
Created by transformation of PCR product of primers MPC70-S2 and MPC70-S3 with pYM12 Monomeric as template into YMJ146.
Checked by test PCR with hiskantag2 and MPC70-tag.
This work
YMM14 MATa/MATα
MPC54-yemGFP::KanMX cyr1Δ::hphNT1/ MPC54-yemGFP::KanMX cyr1Δ::hphNT1
pMJ15 [CYR1 URA3 ARS/CEN]
Created by crossing YMM7 and YMM8.
This work
YMM15 MATa/MATα
ADY1-yemGFP::KanMX cyr1Δ::hphNT1/ ADY1-yemGFP::KanMX cyr1Δ::hphNT1
pMJ15 [CYR1 URA3 ARS/CEN]
Created by crossing YMM10 and YMM11
This work
YMM16 MATα
MPC70-yemGFP::KanMX cyr1Δ::hphNT1 pDS66 [cyr1K1712A URA3 ARS/CEN]
Created by transformation of PCR product of primers MPC70-S2 and MPC70-S3 with pYM12 Monomeric as template into YMJ145.
Checked by test PCR with hiskantag2 and MPC70-tag.
This work
YMM17 MATα
SPO74-yemGFP::KanMX cyr1Δ::hphNT1 pDS66 [cyr1K1712A URA3 ARS/CEN]
Created by transformation of PCR product of primers SPO74-S2 and SPO74-S3 with pYM12 Monomeric as template into YMJ146.
Checked by test PCR with hiskantag2 and SPO74-tag.
This work
YMM18 MATa/MATα This work
89 MPC70-yemGFP::KanMX cyr1Δ::hphNT1/
MPC70-yemGFP::KanMX cyr1Δ::hphNT1 pDS66 [cyr1K1712A URA3 ARS/CEN]
Created by crossing YMM16 and YMM13.
YMM19 MATα
SPO74-yemGFP::KanMX cyr1Δ::hphNT1 pDS66 [cyr1K1712A URA3 ARS/CEN]
Created by transformation of PCR product of primers SPO74-S2 and SPO74-S3 with pYM12 Monomeric as template into YMJ145.
Checked by test PCR with hiskantag2 and SPO74-tag.
This work
YMM20 MATa/MATα
SPO74-yemGFP::KanMX cyr1Δ::hphNT1/ SPO74-yemGFP::KanMX cyr1Δ::hphNT1
pDS66 [cyr1K1712A URA3 ARS/CEN]
Created by crossing YMM19 and YMM17
This work
YMM21 MATα
MPC54-yemGFP::KanMX cyr1Δ::hphNT1 pDS66 [cyr1K1712A URA3 ARS/CEN]
Created by transformation of PCR product of primers MPC54-S2 and MPC54-S3 with pYM12 Monomeric as template into YMJ146.
Checked by test PCR with hiskantag2 and MPC54-tag.
This work
YMM22 MATa
ADY1-yemGFP::KanMX cyr1Δ::hphNT1 pDS66 [cyr1K1712A URA3 ARS/CEN]
Created by transformation of PCR product of primers ADY1-S2 and ADY1-S3 with pYM12 Monomeric as template into YMJ145.
Checked by test PCR with hiskantag2 and ADY1-tag.
This work
YMM23 MATa/MATα
MPC54-yemGFP::KanMX cyr1Δ::hphNT1/ MPC54-yemGFP::KanMX cyr1Δ::hphNT1
pDS66 [cyr1K1712A URA3 ARS/CEN]
Created by crossing YMM9 and YMM21.
This work
YMM24 MATa/MATα
ADY1-yemGFP::KanMX cyr1Δ::hphNT1/ ADY1-yemGFP::KanMX cyr1Δ::hphNT1
pDS66 [cyr1K1712A URA3 ARS/CEN]
Created by crossing YMM22 and YMM12
This work
YMM25 MATa
nrg1Δ::hphNT1
Created by transformation of PCR product of primers NRG1-S1 and NRG1-S2 with pFA61-hphNT1 as template into YMJ36.
Checked by test PCR with hiskantag2 and NRG1-KO
This work
YMM26 MATα
nrg1Δ::hphNT1
This work
90 Created by transformation of PCR product of primers NRG1-S1 and
NRG1-S2 with pFA61-hphNT1 as template into YMJ37.
Checked by test PCR with hiskantag2 and NRG1-KO
YMM27 MATα
yap1Δ::hphNT1
Created by transformation of PCR product of primers YAP1-S1 and YAP1-S2 with pFA61-hphNT1 as template into YMJ37.
Checked by test PCR with hiskantag2 and YAP1-KO
This work
YMM28 MATa/MATα
nrg1Δ::hphNT1/nrg1Δ::hphNT1
Created by crossing YMM25 and YMM26
This work
YMM29 MATa
yap1::hphNT1
Created by transformation of PCR product of primers YAP1-S1 and YAP1-S2 with pFA61-hphNT1 as template into YMJ36.
Checked by test PCR with hiskantag2 and YAP1-KO
This work
YMM30 MATa/MATα
yap1Δ::hphNT1/yap1Δ::hphNT1
Created by crossing YMM27 and YMM29
This work
YMM34 MATa/MATα
ura3-52 lys2 ho::LYS2/leu2delta ura3-52 lys2 his3::IME2(P)-pTEV+DIT1(T)::kanMX4::his3/
his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3 trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1/
trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1
natNT2::SCC1(P)::GFP-TEVdegF-SF3B-3HA/
natNT2::SCC1(P)::GFP-TEVdegF-SF3B-3HA
Created by transformation of PCR product of primers BCY1-S1 and BCY1-S4 with pCR65 as template into YCR329
This work
YMM36 MATa
lys2 leu2 ho::LYS2 ura3
his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3 trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1
NatNT2::Adh1(P)-GFP-TDegF-SF3B-3HA-BCY1
Created by transformation of PCR product of primers BCY1-S1 and BCY1-S4 with pCT289 as template into YAA130.
Checked by test PCR with hiskantag2 and BCY1-KO.
This work
YMM37 MATα
lys2 leu2 ho::LYS2 ura3
his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3 trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1
NatNT2::Adh1(P)-GFP-TDegF-SF3B-3HA-BCY1
Created by transformation of PCR product of primers BCY1-S1 and
This work
91 BCY1-S4 with pCT289 as template into YAA129.
Checked by test PCR with hiskantag2 and BCY1-KO.
YMM40 MATa
snf4Δ::hphNT1
Created by transformation of PCR product of primers SNF4-S1 and SNF4-S2 with pFA61-hphNT1 as template into YMJ36.
Checked by test PCR with hiskantag2 and SNF4-KO
This work
YMM41 MATα snf4::hphNT1
Created by transformation of PCR product of primers SNF4-S1 and SNF4-S2 with pFA61-hphNT1 as template into YMJ37.
Checked by test PCR with hiskantag2 and SNF4-KO
This work
YMM42 MATa
fob1::hphNT1
Created by transformation of PCR product of primers FOB1-S1 and NRG1-S2 with pFA61-hphNT1 as template into YMJ36.
Checked by test PCR with hiskantag2 and FOB1-KO
This work
YMM43 MATα
fob1::hphNT1
Created by transformation of PCR product of primers NRG1-S1 and FOB1-S2 with pFA61-hphNT1 as template into YMJ37.
Checked by test PCR with hiskantag2 and FOB1-KO
This work
YMM45 MATa/MATα
fob1Δ::hphNT1/fob1Δ::hphNT1
Created by crossing YMM42 and YMM43
This work
YMM46 MATa/MATα
lys2/lys2 leu2/leu2 ho::LYS2/ho::LYS2 ura3/ura3 his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3/
his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3 trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1/
trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1 NatNT2::Adh1(P)-GFP-TDegF-SF3B-3HA-BCY1/
NatNT2::Adh1(P)-GFP-TDegF-SF3B-3HA-BCY1
Created by crossing YMM36 and YMM37
This work
YMM50 MATa
pnc1Δ::hphNT1
Created by transformation of PCR product of primers PNC1-S1 and PNC1-S2 with pFA61-hphNT1 as template into YMJ36.
Checked by test PCR with hiskantag2 and PNC1-KO
This work
YMM51 MATα
pnc1Δ::hphNT1
Created by transformation of PCR product of primers PNC1-S1 and PNC1-S2 with pFA61-hphNT1 as template into YMJ37.
Checked by test PCR with hiskantag2 and PNC1-KO
This work
92 YMM52 MATa/MATα
pnc1Δ::hphNT1/pnc1Δ::hphNT1
Created by crossing YMM50 and YMM51
This work
YMM54 MATa
lys2 leu2 ho::LYS2 ura3
his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3 trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1
natNT2::Pcyc1::GFP-TEVdeg-SF3B-3HA-BCY1
Created by transformation of PCR product of primers BCY1-S1 and BCY1-S4 with pMJ8 as template into YAA129.
Checked by test PCR with hiskantag2 and BCY1-KO.
This work
YMM55 MATa/MATα
snf4Δ::hphNT1/snf4Δ::hphNT1
Created by crossing YMM40 and YMM41
This work
YMM60 MATα
lys2 leu2 ho::LYS2 ura3
his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3 trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1
natNT2::Pcyc1::GFP-TEVdeg-SF3B-3HA-BCY1
Created by transformation of PCR product of primers BCY1-S1 and BCY1-S4 with pMJ8 as template into YAA130.
Checked by test PCR with hiskantag2 and BCY1-KO.
This work
YMM64 MATa/MATα
ura3-52 lys2 ho::LYS2/leu2delta ura3-52 lys2 his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3/
his3::IME2(P)-pTEV+-DIT1(T)::kanMX4::his3 trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1/
trp1::IME2(P)-pTEV+-DIT1(T)::kanMX4::trp1
natNT2::Pcyc1::GFP-TEVdeg-SF3B-3HA-BCY1/
natNT2::Pcyc1::GFP-TEVdeg-SF3B-3HA-BCY1
Created by crossing YMM54 and YMM60
This work
YUK63 MATa/MATα
YKS32 SPO74-yeGFP::kanMX/SPO74-yeGFP::kanMX
(Taxis et al., 2005)