Key resources table.
| Reagent type (Species) or resource |
Designation | Source Or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa ade2-1 trp1-1 can1-100 leu2-3,112 his3-11,15 ura3 GAL psi+
All following strains are based on this background |
This study | K699 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa scc2-4 SCC1-PK9 TETR-GFP::LEU2 TET Os::URA3 trp1:MET3p-CDC20::TRP1 | This study | K15028 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae Mata trp1:smc3(E1155Q)-PK6::TRP1 | This study | K17409 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae Mata SCC1-PK9::KAN MX rad61Δ::HGH MX leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4 (single copy) | This study | K20891 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae Mat a MATa SCC1-PK9::KanMX rad61Δ::HGH MX smc3(R1008I)::HIS3 scc2-45::NAT MX (L545P D575G) leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4(single copy) | This study | K22294 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae Mat a MATa SCC1-PK9::KanMX rad61Δ::HGH MX smc3(R1008I)::HIS3 leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4(single copy) | This study | K22296 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa eco1Δ::NatMX4 rad61Δ::HGH MX scc2-45::NAT MX (L545P D575G) Smc3(E199A/R1008I)::HIS37.5 kb minichromosome (TRP1 ARS1 CEN1-6KB) | This study | K22297 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa eco1Δ::NatMX4 rad61Δ::HGH MX Smc3(E199A/R1008I)::HIS3 7.5 kb minichromosome (TRP1 ARS1 CEN1-6KB) | This study | K22298 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa scc1(A547C)-PK6::KanMX smc3(E570C,S1043C)::ADE2 smc1(G22C,K639C)::NatMX2.3 kb Trp1-ARS1-Cen4 plasmid | Srinivasan et al. (2018) | K23889 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa scc1(A547C)-PK6::KanMX smc3(E570C,S1043C)::ADE2 smc1(G22C,K639C)::NatMX leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4
(single copy) 2.3 kb Trp1-ARS1-Cen4 plasmid |
Srinivasan et al. (2018) | K23972 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa Scc1(A547C)-pk6::KanMX Smc3(E570C,S1043C)::ADE2 Smc1(G22c,K639C)::NatMXscc2-45::natMX (L545P D575G)
2.3 kb Trp1-ARS1-Cen4 plasmid |
Srinivasan et al. (2018) | K24267 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa Smc1(G22C,K639C)::NatMX4
Smc3(E570C,S1043C)::ADE2 leu2::Gal-Scc1(R180E,R268D, A547C)-PK6::LEU2 SCC3 (K404E)-HA3::HIS 2.3 kb Trp1-ARS1-Cen4 plasmid |
This study | K24697 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa Scc1(A547C)-pk6::KanMX
Smc3(E570C,S1043C)::ADE2 Smc1(G22c,K639C)::NatMX scc2-45::natMX (L545P D575G) scc3 (K404E)-HA3::HIS 2.3 kb Trp1-ARS1-Cen4 plasmid |
This study | K24709 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa Smc1(G22C,K639C)::NatMX4
Smc3(E570C,S1043C)::ADE2 leu2::Gal-Scc1(R180E,R268D, A547C)-PK6::LEU2 scc3 (K404E)-HA3::HIS scc2-45::natMX (L545P D575G) TRP1-ARS1-CEN4 2.3 KB plasmid |
This study | K24738 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa trp1::smc3(E1155Q)-
PK6::TRP1 smc1(D1164E) |
This study | K25039 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa Scc1(A547C)-pk6::KanMX Smc3(E570C,S1043C)::ADE2 Smc1(G22c,K639C)::NatMX scc2-45::natMX (L545P D575G) 2.3 kb Trp1-ARS1-Cen4 plasmid | This study | K25238 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa scc1(A547C)-PK6::KanMX pds5 (s81R)::HIS3 smc3(E570C,S1043C)::ADE2 smc1(G22C,K639C)::NatMX leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4
(single copy) 2.3 kb Trp1-ARS1-Cen4 plasmid |
This study | K25311 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa scc1(A547C)-PK6::KanMX pds5 (s81R)::HIS3 smc3(E570C,S1043C)::ADE2 smc1(G22C,K639C)::NatMX scc2-45::natMX (L545P D575G) leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4 (single copy) 2.3 kb Trp1-ARS1-Cen4 plasmid | This study | K25312 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa scc1(A547C)-PK6::KanMX scc3 (K404e)-HA3::HIS3
smc3(E570C,S1043C)::ADE2 smc1(G22C,K639C)::NatMX leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4 (single copy) 2.3 kb Trp1-ARS1-Cen4 plasmid |
This study | K25313 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa scc1(A547C)-PK6::KanMX scc3 (K404e)-HA3::HIS3 smc3(E570C,S1043C)::ADE2 smc1(G22C,K639C)::NatMX scc2-45::natMX (L545P D575G) leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4 (single copy) 2.3 kb Trp1-ARS1-Cen4 plasmid | This study | K25316 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa scc1(A547C)-PK6::KanMX smc3(E570C,S1043C)::ADE2 smc1(G22C,K639C)::NatMX scc2-45::natMX (L545P D575G) cdc28-as1 | This study | K25423 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa smc3(E570C,S1043C)::ADE2 smc1(G22C,K639C)::NatMX leu2::Gal-Scc1(R180E,R268D, A547C)-PK6::LEU2 scc3 (K404e)-HA3::HIS3 cdc28-as1 2.3 kb Trp1-ARS1-Cen4 plasmid | This study | K25437 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa smc3(E570C,S1043C)::ADE2 smc1(G22C,K639C)::NatMX leu2::Gal-Scc1(R180E,R268D, A547C)-PK6::LEU2 scc2-45::natMX (L545P D575G) scc3 (K404e)-HA3::HIS3 cdc28-as1 2.3 kb Trp1-ARS1-Cen4 plasmid | This study | K25440 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa smc3(E570C,S1043C)::ADE2 smc1(G22C,K639C)::NatMX scc1(A547C)-PK6::KanMX scc2-45::natMX (L545P D575G) eco1Δ::HGH MX scc3 (K404E)-HA3::HIS TRP1-ARS1-CEN4 2.3 KB plasmid | This study | K25947 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa ura::ADH1 promoter-OsTIR1-9myc::URA3 PDS5-Pk3-aid::KanMX4scc2-45::NatMX scc3 (K404E)::HA3::HIS leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4::Leu2 (single copy) | This study | K26414 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa ura::ADH1 promoter-OsTIR1-9myc::URA3 PDS5-Pk3-aid::KanMX4 scc3 (K404E)::HA3::HIS leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4::Leu2 (single copy) | This study | K26415 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa smc1(D1164E)::HIS3MX
SCC1-PK9::KanMX leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4::Leu2 (single copy) |
This study | K26765 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa smc1(D1164E)::HIS3MX
SCC1-PK9::KanMX scc2-45::natMX (L545P D575G) leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/ Gal1p-Gal4::Leu2 (single copy) |
This study | K26766 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa ura::ADH1 promoter-OsTIR1-9myc::URA3 scc2-3XmAID::KANMX6 Scc1-PK9::KanMX rad61Δ::HGH MX leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4::Leu2 (single copy) | This study | K26831 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa scc2-45::natMX (L545P D575G) leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4::Leu2 (single copy) ura3::Scc1P-Smc3(E570C)-TEV3-Scc1(A547C)-PK9::KanMX::URA3 (single integrant,fusion linker: (GGGGS)x8 + TEV3) | This study | K26993 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa scc2-45::natMX (L545P D575G) leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4::Leu2 (single copy) Smc1(G22C, K639C)::NAT MX ura3::Scc1P-Smc3(E570C)-TEV3-Scc1(A547C)-PK9::KanMX::URA3 (single integrant,fusion linker: (GGGGS)x8 + TEV3) | This study | K26994 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa bar1::hisG ura3-1::GAL-OsTIR1-9Myc::URA3 cdc45-AID (CDC45-5xGA-IAA17)::kanMX rad61-3slAA::natNT2 SCC1-PK6::TRP1 scc2-45::natMX (L545P D575G) | This study | K27168 | Materials and methods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa bar1::hisG ura3-1::GAL-OsTIR1-9Myc::URA3 cdc45-AID (CDC45-5xGA-IAA17)::kanMX rad61-3slAA::natNT2 SCC1-PK6::TRP1 | This study | K27169 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa TET R-GFP::LEU2
TETOs::URA3 pds5 S81R-HA6::HIS (NO HA tag) trp1:MET3p-CDC20::TRP1 |
This study | K27443 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa SCC1-PK9::KanMX smc3(T1185M)-HIS3M × 6 scc2-45::natMX (L545P D575G) leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4::Leu2 (single copy) | This study | K27536 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa SCC1-PK9::KanMX smc3(T1185M)-HIS3M × 6 leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4::Leu2 (single copy) | This study | K27537 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa SCC1-PK9::KanMX rad61Δ::hghMX leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4::Leu2 (single copy) | This study | K27569 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) | S. cerevisiae MATa SCC1-PK9::KanMX rad61Δ::hghMX scc4-2:NAT leu2::Gal1p-Sic1(9 m)/His3p-Gal1/His3p-Gal2/Gal1p-Gal4::Leu2 (single copy) | This study | K27570 | Materials andmethods subsection experimental models |
| Strain, strain background (S. cerevisiae) |
S. cerevisiae MATa scc2-4 TET R-GFP::LEU2
TETOs::URA3 pds5 S81R-HA6::HIS (NO HA tag) trp1:MET3p-CDC20::TRP1 |
This study | K27575 | Materials andmethods subsection experimental models |
| Strain, strain background (Spodoptera frugiperda) | Sf9 insect cells | ThermoFisher | Cat# 11496015 | |
| Antibody | Mouse monoclonal Anti-V5 | BioRad | Cat# MCA1360 | (1:1000) |
| Antibody | Mouse monoclonal Anti Smc3 Acetyl | Beckouët et al. (2010) | N/A | (1:1000) |
| Antibody | Rabbit polyclonal Anti Clb2 y-180 | SantaCruz | Sc-9071 | (1:1000) |
| Antibody | Rabbit polyclonal Anti Sic1 FL-284 | SantaCruz | Sc-50441 | (1:1000) |
| Antibody | Mouse monoclonal Anti PGK1 | ThermoFisher Scientific | Cat#459250 | (1:5000) |
| Antibody | Rabbit polyclonal Anti-mini-AID-tag mAb | Kanemaki MT/MBL International | M214-3 | (1:1000) |
| Chemical compound, drug | Acid-washed glass beads | Sigma | Cat# G8722 | |
| Chemical compound, drug | ATP α-32P | Hartmann Analytic | Cat# SRP-203 | |
| Chemical compound, drug | Bismaleimidoethane (BMOE) | ThermoFisher | Cat# 22323 | (5 mM) |
| Chemical compound, drug | Complete EDTA free protease inhibitor cocktail | Roche | Cat# 4693132001 | (1:50 ml) |
| Chemical compound, drug | Dithiothreitol | Fluka | Cat# BP172 | (5 mM) |
| Chemical compound, drug | DMSO | Sigma | Cat# D8418 | |
| Chemical compound, drug | Immobilon Western ECL | Millipore | Cat# WBLKS0500 | |
| Chemical compound, drug | Trisodium citrate | Sigma | Cat# W302600 | |
| Chemical compound, drug | RNase A | Roche | Cat# 10109169001 | |
| Chemical compound, drug | Nocodazole | Sigma | Cat# M1404 | |
| Chemical compound, drug | 1NMPP1 | MerkMillipore | 529581 | |
| Chemical compound, drug | Indole-3-acetic acid (auxin) | Sigma | Cat# I3750-5G-A | |
| Chemical compound, drug | PMSF | Sigma | Cat# 329-98-6 | |
| Chemical compound, drug | Potassium chloride | Sigma | Cat# P5405 | |
| Chemical compound, drug | Proteinase K | Roche | Cat# 03115836001 | |
| Chemical compound, drug | Sodium sulfite | Sigma | Cat# 71988 | |
| Peptide, recombinant protein | α-factor peptide | CRUK Peptide Synthesis Service | ||
| Biological sample (Saccharomyces cerevisiae) | Saccharomyces cerevisiae cohesin | This study | Materials and methods subsection Tetramer and Scc2 purification | |
| Biological sample (Saccharomyces cerevisiae) | Saccharomyces cerevisiae Scc2 | This study | Materials andmethods subsection Tetramer and Scc2 purification | |
| Commercial assay or kit | AxyPrep Mag PCR Clean up Kit | Appleton Woods Ltd. | Cat# AX402 | |
| Commercial assay or kit | ChIP Clean and Concentrator Kit | Zymo Research | Cat# D5205 | |
| Commercial assay or kit | E-Gel SizeSelect II Agarose Gels, 2% | ThermoFisher | Cat# G661012 | |
| Commercial assay or kit | EnzChek phosphate assay kit | Invitrogen | Cat# E6646 | |
| Commercial assay or kit | HiTrap TALON column | GE Healthcare | Cat# 28-9537-67 | |
| Commercial assay or kit | Library Quantification Kit Ion Torrent Platforms | KAPA Biosystems | Cat# KR0407 | |
| Commercial assay or kit | Microcon YM-100 columns | Sigma | Cat# Z648094 | |
| Commercial assay or kit | NEBNext Fast DNA library prep set for Ion Torrent | NEB | Cat# E6270L | |
| Commercial assay or kit | NuPAGE 3–8% Tris-Acetate Protein Gels, 1.5 mm, 10-well | ThermoFisher | Cat# EA0378BOX | |
| Commercial assay or kit | NuPAGE 4–12% Bis-Tris Protein Gels, 1.0 mm, 10-well | ThermoFisher | Cat# NP0321BOX | |
| Commercial assay or kit | Prime-it II Random Primer Labelling Kit | Agilent | Cat# 300385 | |
| Commercial assay or kit | Protein G dynabeads | ThermoFisher | Cat# 10003D | |
| Commercial assay or kit | Slide-a-lyzer dialysis units (3.5 kDa) | ThermoFisher | Cat# 66330 | |
| Commercial assay or kit | StrepTrap HP column | GE Healthcare | Cat# 28-9075-48 | |
| Commercial assay or kit | Superdex 200 16/60 GL | GE Healthcare | Cat# 17-1069-01 | |
| Commercial assay or kit | Superose 6 10/300 GL | GE Healthcare | Cat# 17517201 | |
| Commercial assay or kit | TALON Superflow metal affinity resin | Clontech | Cat# 635670 | |
| Software, algorithm | Galaxy platform | Giardine et al. (2005) | https://usegalaxy.org | |
| Software, algorithm | FastQC | Galaxy tool version 1.0.0 |
https://usegalaxy.org | |
| Software, algorithm | Trim sequences | Galaxy tool version 1.0.0 |
https://usegalaxy.org | |
| Software, algorithm | Filter FASTQ | Galaxy tool version 1.0.0 | https://usegalaxy.org | |
| Software, algorithm | Bowtie2 |
Langmead and Salzberg (2012)
Galaxy tool version 0.2 |
https://usegalaxy.org | |
| Software, algorithm | Bam to BigWig | Galaxy tool version 0.1.0 | https://usegalaxy.org | |
| Software, algorithm | Samtools | Li et al. (2009) | http://samtools.sourceforge.net/ | |
| Software, algorithm | IGB browser | Nicol et al. (2009) | http://bioviz.org/igb/ | |
| Software, algorithm | Filter SAM or BAM |
Li et al. (2009)
Galaxy tool version 1.1.0 |
https://usegalaxy.org | |
| Software, algorithm | chr_position.py | This study | https://github.com/naomipetela/nasmythlab-ngs | |
| Software, algorithm | filter.py | This study | https://github.com/naomipetela/nasmythlab-ngs | |
| Software, algorithm | bcftools call | Li et al. (2009) |