KEY RESOURCES TABLE
REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Secondary Goat Anti-Mouse Immunoglobulins/HRP, 1:5000 | Dako | P0447; RRID:AB_2617137 |
Goat Anti-Rabbit Immunoglobulins/HRP, WB:1:5000 | Dako | P0448;: RRID:AB_2617138 |
Anti-HA (anti-HA.11 Epitope Tag Antibody clone 16B12), WB:1:2000‘ | BioLegend | 901502, lot B276381 or B242905; RRID:AB_2565006 |
Anti-Pgk1 antibody [22C5D8], WB:1:3000 | Abcam | ab113687, GR3195775-3; RRID:AB_10861977 |
Anti-Rad53 (monoclonal, K-medium), WB:1:100 | Gift from Maksym Shyian (D. Shore laboratory) | Mab clone EL7.E1 |
Anti-Flag M2, WB:1:5000 | Sigma | F3165, lot SLBQ7119V or SLCC4005: RRID:AB259529 |
Anti-Myc, WB: 1:5000 | Abcam | Ab32; lot GR310953-2 or GR3232358-2; RRID:AB_303599 |
Anti-Ubiquitin FK2 | Calbiochem | ST1200-100UG, lots 31671118, 3291266, 3443858; RRID:AB_2043482 |
Anti-SUMO (polyclonal), WB: 1:10000 | Helle Ulrich laboratory: (Papouli et al., 2005) | N/A |
Anti-PCNA (polyclonal), WB: 1:2500 | Helle Ulrich laboratory: (Papouli et al., 2005) | N/A |
Bacterial and virus strains | ||
E. coli DH5α | N/A | N/A |
Chemicals, peptides, and recombinant proteins | ||
D(+)-Glucose | Carl Roth | Art.-Nr. HN063 |
D(+)-Raffinose pentahydrate | Carl Roth | Art.-Nr. 5241.3 |
D(+)-Galactose | USBiological | G1030 |
Yeast Nitrogen Base | Qbiogene | 4027-532 |
Bacto Peptone | BD Biosciences | Ref. 211677 |
Yeast Extract | BD Biosciences | Ref. 212750 |
Ammonium Sulfate | Carl Roth | Art.-Nr. 3746.1 |
5-Fluoroorotic Acid | Toronto Research Chemicals | F595000 |
RNase A, PureLink, (20 mg/mL) | Invitrogen | 12091-021 |
DpnII | NEB | R0543L |
NlaIII | NEB | R0125L |
T4 Ligase | Thermo Scientific | EL0011 |
Linear Acrylamide | Ambion | AM9520 |
Taq polymerase | NEB | M0267L |
Salmon sperm ssDNA | Sigma-Aldrich | D1626 |
PEG4000 | Carl Roth | Art.-Nr. 0156.1 |
Zymolyase - 20T | Amsbio | 120491-1 |
Phusion High-Fidelity DNA Polymerase | Thermo Scientific | F530L |
Auxin (3-indoleacetic acid) | Sigma-Aldrich | I2886 |
Camptothecin (CPT) | Lucerna-Chem | Cat. 0215973225 |
WesternBright ECL HRP substrate | Advansta | K-12045 |
WesternBright Sirius HRP substrate | Advansta | K-12043 |
cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail | Roche | 4693159001 |
NEM (N-Ethylmaleimide) | Sigma-Aldrich | E1271 |
Dynabeads Protein G for Immunoprecipitation | Invitrogen | 10009D |
Dynabeads Pan Mouse IgG | Invitrogen | 11041 |
Alpha Factor | PRIMM | 201307-00007 |
Trypsin | Promega | V5113 |
Propidium Iodide | Sigma | P4170 |
Proteinase K | Carl Roth | 7528.3 |
Ni-NTA agarose | QIAGEN | 30210 |
Critical commercial assays | ||
The Bio-Rad protein assay | Bio-Rad | 500-0006 |
Phire Green Hot Start II PCR Master Mix | Thermo Scientific | F126L |
NEBuilder® HiFi DNA Assembly Cloning Kit | NEB | E5520 |
NucleoSpin® Gel and PCR Clean-up | Macherey-Nagel | Ref. 740609.250 |
MiSeq Reagent Kit v3 | Illumina | MS-102 |
SYBR Select Master Mix for CFX | Applied biosystems | 4472942 |
MinElute PCR Purification Kit | QIAGEN | 28006 |
Deposited data | ||
Sequencing data of the tdp1Δ wss1Δ siz2Δ transposon screen | This Manuscript | ENA: PRJEB47156 |
Sequencing data of “wss1D Tdp1-AID*-6HA TIR1 + auxin” transposon screen | Serbyn et al. (2020), Mol.Cell | ENA: PRJEB31382 |
The SUMO mass spectrometry proteomics data | This Manuscript | MassIVE: MSV000088129 |
Unprocessed imaging (immunoblots, spot assays, tetrad analyses) and FACS data | This Manuscript | Mendeley Data: https://data.mendeley.com/datasets/h7bxnynd7s/1 |
Experimental models: Organisms/strains | ||
S. cerevisiae: Strain background: W303 | EUROSCARF | BMA64 |
S. cerevisiae: Strain background: BY4741 | EUROSCARF | Y00000 |
Oligonucleotides | ||
See Table S3 for the list of oligonucleotides | This Manuscript | N/A |
Recombinant DNA | ||
See Table S3 for full list of plasmids | This Manuscript | N/A |
Software and algorithms | ||
Prism 9.1.2 | GraphPad Software, Inc | RRID:SCR_002798 |
RStudio Version 1.1.456 | RStudio, Inc | RRID:SCR_000432 |
Fiji | ImageJ | RRID:SCR_002285 |
SnapGene 3.2.1 | GSL Biotech LLC | RRID:SCR_015052 |
Kaluza | Beckman Coulter | RRID:SCR_016182 |
Other | ||
0.5 mm Glass Beads | BioSpec Products | Cat. No 11079105 |
MagNA Lyser Instrument | Roche | 03358976001 |
Bioruptor Twin | Diagenode | UCD-400 |
Gallios 8 colors 2 Lasers flow cytometer | Beckman Coulter | B43619 |