REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Mouse monoclonal anti V5-tag (anti PK6-tag) | AbD Serotec (MCA1360) | RRID: AB_322378 |
Mouse monoclonal anti tubulin (TAT1) | Woods et al., 1989 | N/A |
Rabbit polyclonal anti Sc Ycg1 | Piazza et al., 2014 | N/A |
Bacterial and Virus Strains | ||
Escherichia coli Rosetta (DE3) pLysS | Merck | Cat#70954 |
Chemicals, Peptides, and Recombinant Proteins | ||
Crosslinker dibromobimane (bBBr) | Sigma-Aldrich | Cat#34025 |
Crosslinker bismaleimidoethane (BMOE) | Thermo-Fisher | Cat#22323 |
Crosslinker disthiobismaleimidoethane (DTME) | Thermo-Fisher | Cat#22335 |
C. thermophilum Ycs4 (3-1222, N-terminal 6 × HIS-tag) | modified purification protocol from Piazza et al., 2014 | N/A |
C. thermophilum Ycg1 (24-1006, N-terminal 6 × HIS-tag) | modified purification protocol from Piazza et al., 2014 | N/A |
C. thermophilum Brn1336-714 (336-714, N-terminal GST-tag) | This work | N/A |
C. thermophilum Brn1515-634 (515-634, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-634, BC1 (515-634, R539D, R541D, K542D, K544D, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-634, BC2 (515-634, R554D, R556D, K557D, K559D, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-634, BC1/2 (515-634, R539D, R541D, K542D, K544D, R554D, R556D, K557D, K559D, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-634 (515-634, N-terminal 6 × HIS-tag) in complex with Ycg1YC1 (24-1006, K100D, K101D) | This work | N/A |
C. thermophilum Brn1515-634 (515-634, N-terminal 6 × HIS-tag) in complex with Ycg1YC2 (24-1006, K916D, K917D) | This work | N/A |
C. thermophilum Brn1515-634 (515-634, N-terminal 6 × HIS-tag) in complex with Ycg1YC1/2 (24-1006, K100D, K101D, K916D, K917D) | This work | N/A |
C. thermophilum Brn1225-512 (225-512, N-terminal 6 × HIS-tag) in complex with Ycs4 (3-1222) | This work | N/A |
C. thermophilum Brn1539-634 (539-634, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1549-634 (549-634, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1572-634 (572-634, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-601 (515-601, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-634 (515-634, N-terminal 6 × HIS-tag) in complex with Ycg178-1006 (78-1006) | This work | N/A |
C. thermophilum Brn1515-634 (515-634, N-terminal 6 × HIS-tag) in complex with Ycg124-823 (24-823) | This work | N/A |
C. thermophilum Brn1515-634 (515-634, N-terminal 6 × HIS-tag) in complex with Ycg124-883 (24-883) | This work | N/A |
C. thermophilum Brn1515-634 (515-634, N-terminal 6 × HIS-tag) in complex with Ycg124-934 (24-934) | This work | N/A |
C. thermophilum Brn1515-634 (515-634, N-terminal 6 × HIS-tag) in complex with Ycg124-982 (24-982) | This work | N/A |
C. thermophilum Brn1515-634, E514C-R629C (515-634, E514C, R629C, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-634, E514C-R629C cleavable (515-634, E514C, R629C, N-terminal 6 × HIS-tag, HRV3C cleavage site inserted after P549) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-634, L521C-S611C cleavable (515-634, L521C, S611C, N-terminal 6 × HIS-tag, HRV3C cleavage site inserted after P549) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-634, D525C-S610C cleavable (515-634, D525C, S610C, N-terminal 6 × HIS-tag, HRV3C cleavage site inserted after P549) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-634, E514C-Q613C cleavable (515-634, E514C, Q613C, N-terminal 6 × HIS-tag, HRV3C cleavage site inserted after P549) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-634, A527C-R629C cleavable (515-634, A527C, R629C, N-terminal 6 × HIS-tag, HRV3C cleavage site inserted after P549) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-634, S568C-R629C cleavable (515-634, S568C, R629C, N-terminal 6 × HIS-tag, HRV3C cleavage site inserted after P549) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1515-634, cleavable (515-634, N-terminal 6 × HIS-tag, HRV3C cleavage site inserted after P549) in complex with Ycg1 (24-1006) | This work | N/A |
C. thermophilum Brn1225-634 (225-634, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) and Ycs4 (3-1222) | This work | N/A |
C. thermophilum Brn1225-634 (225-634, N-terminal 6 × HIS-tag) in complex with Ycg1YC1+YC2 (24-1006, K100D, K101D, K916D, K917D) and Ycs4 (3-1222) | This work | N/A |
C. thermophilum Brn1225-634 BC1/2 (225-634, R539D, R541D, K542D, K544D, R554D, R556D, K557D, K559D, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) and Ycs4 (3-1222) | This work | N/A |
C. thermophilum Brn1225-634, E514C-R629C (225-634, E514C, R629C, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) and Ycs4 (3-1222) | This work | N/A |
C. thermophilum Brn1225-634, E514C-R629C cleavable (225-634, E514C, R629C, N-terminal 6 × HIS-tag, HRV3C cleavage site inserted after P549) in complex with Ycg1 (24-1006) and Ycs4 (3-1222) | This work | N/A |
C. thermophilum Brn1225-634, ΦD (225-634, L521D, F524D, W532D, W538D, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) and Ycs4 (3-1222) | This work | N/A |
C. thermophilum Brn1515-634, ΦD (515-634, L521D, F524D, W532D, W538D, N-terminal 6 × HIS-tag) in complex with Ycg1 (24-1006) | This work | N/A |
S. cerevisiae Smc2 (full length) in complex with Smc4 (full length, C-terminal 3 × StrepII-tag), Brn1 (full length, C-terminal 12 × HIS-tag 3 × HA-tag), Ycg1 (full length) and Ycs4 (full length) | Terekawa et al., 2017 | N/A |
S. cerevisiae Smc2 (full length) in complex with Smc4 (full length, C-terminal 3 × StrepII-tag), Brn1 (full length, C-terminal 12 × HIS-tag 3 × HA-tag) and Ycs4 (full length) | This work | N/A |
S. cerevisiae Smc2 (full length) in complex with Smc4 (full length, C-terminal 3 × StrepII-tag), Brn1BC1/2 (full length, K409D, R411D, K414D, K451D, K452D, K454D, K456D, K457D, C-terminal 12 × HIS-tag 3 × HA-tag), Ycg1 (full length) and Ycs4 (full length) | This work | N/A |
S. cerevisiae Smc2 (full length) in complex with Smc4 (full length, C-terminal 3 × StrepII-tag), Brn1ΦD (full length, M391D, F394D, W402D, W408D, C-terminal 12 × HIS-tag 3 × HA-tag), Ycg1 (full length) and Ycs4 (full length) | This work | N/A |
S. cerevisiae Smc2Q147L (full length) in complex with Smc4Q302L (full length, C-terminal 3 × StrepII-tag), Brn1 (full length, C-terminal 12 × HIS-tag 3 × HA-tag), Ycg1 (full length) and Ycs4 (full length) | Terekawa et al., 2017 | N/A |
S. cerevisiae Brn1 (384-529) Ycg1 (6-498, 556-932) | This work | N/A |
S. cerevisiae Brn1short kleisin loop (384-417, 445-529) Ycg1 (6-498, 556-932) | This work | N/A |
S. pombe Cnd2 (416-544) Cnd3 (1-438, 474-823) | This work | N/A |
Deposited Data | ||
S. pombe Cnd3–Cnd2 | This work | PDB: 5OQR |
S. cerevisiae Ycg1–Brn1 | This work | PDB: 5OQQ |
S. cerevisiae Ycg1–Brn1–DNA (I) | This work | PDB: 5OQP |
S. cerevisiae Ycg1–Brn1–DNA (II) | This work | PDB: 5OQO |
S. cerevisiae Ycg1–Brn1–DNA (short kleisin loop) | This work | PDB: 5OQN |
Experimental Models: Cell Lines | ||
HeLa Kyoto H2B-mCherry | Neumann et al., 2010 | N/A |
Experimental Models: Organisms/Strains | ||
S. cerevisiae C4237 (MATa/α, ade2-1, trp1-1, can1-100, leu2-3,112, GAL, psi+, brn1::HIS3/BRN1, ura3::empty-vector::URA3/ura3) | This work | N/A |
S. cerevisiae C4239 (MATa/α ade2-1, trp1-1, can1-100, leu2-3,112, GAL, psi+, brn1::HIS3/BRN1, ura3::BRN1-PK6::URA3/ura3) | This work | N/A |
S. cerevisiae C4257 (MATa/α ade2-1, trp1-1, can1-100, leu2-3,112, GAL, psi+, brn1::HIS3/BRN1, ura3::brn1(K409D, R411D, K414D)-PK6::URA3/ura3) | This work | N/A |
S. cerevisiae C4259 (MATa/α ade2-1, trp1-1, can1-100, leu2-3,112, GAL, psi+, brn1::HIS3/BRN1, ura3::brn1(K451D, K452D, K454D, K456D, K457D)-PK6::URA3/ura3) | This work | N/A |
S. cerevisiae C4261 (MATa/α ade2-1, trp1-1, can1-100, leu2-3,112, GAL, psi+, brn1::HIS3/BRN1, ura3::brn1(K409D, R411D, K414D, K451D, K452D, K454D, K456D, K457D)-PK6::URA3/ura3) | This work | N/A |
S. cerevisiae C4491 (MATa, lys2::pGAL1 GAL4::LYS2, pep4::HIS3, bar1::hisG, [2micron pGAL7 SMC4-StrepII3, pGAL10 SMC2, pGAL1 BRN1-His12-HA3, TRP1], [2micron pGAL1 YCG1, pGAL10 YCS4, URA3]) | Terekawa et al., 2017 | N/A |
S. cerevisiae C4493 (MATa, lys2::pGAL1 GAL4::LYS2, pep4::HIS3, bar1::hisG, [2micron pGAL7 SMC4-StrepII3, pGAL10 SMC2, pGAL1 BRN1-His12-HA3, TRP1], [2micron pGAL10 YCS4, URA3]) | This work | N/A |
S. cerevisiae C4516 (MATa, lys2::pGAL1 GAL4::LYS2, pep4::HIS3, bar1::hisG, [2micron pGAL7 SMC4-StrepII3, pGAL10 SMC2, pGAL1 BRN1(K409D, R411D, K414D, K451D, K452D, K456D, K457D)-His12-HA3, TRP1], [2micron pGAL1 YCG1, pGAL10 YCS4, URA3]) | This work | N/A |
S. cerevisiae C4724 (MATa, lys2::pGAL1 GAL4::LYS2, pep4::HIS3, bar1::hisG, [2micron pGAL7 SMC4(Q302L)- StrepII3, pGAL10 SMC2(Q147L), pGAL1 BRN1-His12-HA3, TRP1], [2micron pGAL1 YCG1, pGAL10 YCS4, URA3]) | Terekawa et al., 2017 | N/A |
S. cerevisiae C4895 (MATa/α ade2-1, trp1-1, can1-100, leu2-3,112, GAL, psi+, brn1::HIS3/BRN1, ura3::brn1(M391D, F394D, W402D, W408D)-PK6::URA3/ura3) | This work | N/A |
S. cerevisiae C5037 (MATa, lys2::pGAL1 GAL4::LYS2, pep4::HIS3, bar1::hisG, [2micron pGAL7 SMC4-StrepII3, pGAL10 SMC2, pGAL1 BRN1(M391D, F394D, W402D, W408D)-His12-HA3, TRP1], [2micron pGAL1 YCG1, pGAL10 YCS4, URA3]) | This work | N/A |
Oligonucleotides | ||
EMSA 35-mer template forward strand 5′-6-FAM-CCTATAGTGAGTCGTTCGATATTACAATTC ACTGG-3′ |
This work/modified from Piazza et al., 2014 | N/A |
EMSA 35-mer template reverse strand 5′-CCAGTGAATTGTAATATCGAACGACTCACTATAG G-3′ |
This work/modified from Piazza et al., 2014 | N/A |
ITC 25-mer annealed forward strand 5′-CCTATAGTGAGTCACAATTCACTGG-3′ |
This work/modified from Piazza et al., 2014 | N/A |
ITC 25-mer annealed reverse strand 5′-CCAGTGAATTGTGACTCACTATAGG-3′ |
This work/modified from Piazza et al., 2014 | N/A |
Crystallization 18-mer annealed palindromic dsDNA 5′-GATGTGTAGCTACACATC-3′ |
This work | N/A |
22-mer annealed palindromic dsDNA 5′-GATTCGTGTAGCTACACGAATC-3′ |
Seifert et al., 2016 | N/A |
qPCR CEN4 (SC-77) forward primer 5′-TGGTGTGGAAGTCCTAATATCG-3′ |
Cuylen et al., 2011 | N/A |
qPCR CEN4 (SC-78) reverse primer 5′-TGCATGATCAAAAGGCTCAA-3′ |
Cuylen et al., 2011 | N/A |
qPCR rDNA (SC-41) forward primer 5′-TTTCTGCCTTTTTCGGTGAC-3′ |
Cuylen et al., 2011 | N/A |
qPCR CEN4 (SC-42) reverse primer 5′-TGGCATGGATTTCCCTTTAG-3′ |
Cuylen et al., 2011 | N/A |
Recombinant DNA | ||
Plasmid pGEX6PI-Brn1336-714 (N-terminal GST-tag-HRV3C-cleavage-site, residues 336-714 of C. thermophilum Brn1) | This work | N/A |
Plasmid pETMCN-Ycs4 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 3-1222 of C. thermophilum Ycs4) | This work | N/A |
Plasmid pETMCN-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 24-1006 of C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634, BC1 -Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 with mutated residues R539D, R541D, K542D, K544D of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634, BC2 -Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 with mutated residues R554D, R556D, K557D, K559D of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634, BC1/2 -Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 with mutated residues R539D, R541D, K542D, K544D, R554D, R556D, K557D, K559D of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634 -Ycg1YC1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 of C. thermophilum Brn1 and residues 24-1006 with mutated residues K100D, K101D of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634 -Ycg1YC2 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 of C. thermophilum Brn1 and residues 24-1006 with mutated residues K916D, K917D of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634 -Ycg1YC1+YC2 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 of C. thermophilum Brn1 and residues 24-1006 with mutated residues K100D, K101D, K916D, K917D of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1225-512-Ycs4 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 225-512 of C. thermophilum Brn1 and residues 3-1222 of untagged C. thermophilum Ycs4) | This work | N/A |
Plasmid pETMCN-Brn1539-634-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 539-634 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1549-634-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 549-634 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1572-634-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 572-634 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-601-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-601 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634-Ycg178-1006 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 of C. thermophilum Brn1 and residues 78-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634-Ycg124-823 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 of C. thermophilum Brn1 and residues 24-823 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634-Ycg124-883 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 of C. thermophilum Brn1 and residues 24-883 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634-Ycg124-934 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 of C. thermophilum Brn1 and residues 24-934 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634-Ycg124-982 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 of C. thermophilum Brn1 and residues 24-982 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634, E514C-R629C cleavable-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 with mutated residues E514C, R629C and HRV3C cleavage site inserted after P549 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634, L521C-S611C cleavable-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 with mutated residues L521C, S611C and HRV3C cleavage site inserted after P549 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634, D525C-S610C cleavable-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 with mutated residues D525C, S610C and HRV3C cleavage site inserted after P549 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634, E514C-Q613C cleavable-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 with mutated residues E514C, Q613C and HRV3C cleavage site inserted after P549 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634, A527C-R629C cleavable-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 with mutated residues A527C, R629C and HRV3C cleavage site inserted after P549 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634, S568C-R629C cleavable-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 with mutated residues S568C, R629C and HRV3C cleavage site inserted after P549 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634, E514C-R629C-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 with mutated residues E514C, R629C of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634, cleavable-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 with HRV3C cleavage site inserted after P549 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1225-634-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 225-634 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1225-634-Ycg1YC1+YC2 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 225-634 of C. thermophilum Brn1 and residues 24-1006 with mutated residues K100D, K101D, K916D, K917D of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1225-634, BC1/2-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 225-634 with mutated residues R539D, R541D, K542D, K544D, R554D, R556D, K557D, K559D of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1225-634, E514C-R629C-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 225-634 with mutated residues E514C, R629C of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1225-634, E514C-R629C cleavable-Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 225-634 with mutated residues E514C, R629C and HRV3C cleavage site inserted after P549 of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1225-634, ΦD -Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 225-634 with mutated residues L521D, F524D, W532D, W538D of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1515-634, ΦD -Ycg1 (N-terminal 6 × HIS-tag-TEV-cleavage-site, residues 515-634 with mutated residues L521D, F524D, W532D, W538D of C. thermophilum Brn1 and residues 24-1006 of untagged C. thermophilum Ycg1) | This work | N/A |
Plasmid 2micron pGAL7 SMC4-StrepII3 (C-terminal 3 × StrepII-tag of S. cerevisiae full length Smc4), pGAL10 SMC2 (S. cerevisiae full length Smc2), pGAL1 BRN1-His12-HA3 (C-terminal 12 × HIS-3 × HA-tag of S. cerevisiae full length Brn1), TRP1 | St-Pierre et al., 2009 | N/A |
Plasmid 2micron pGAL7 SMC4-StrepII3 (C-terminal 3 × StrepII-tag with mutated residue Q302L of S. cerevisiae full length Smc4), pGAL10 SMC2 (with mutated residue Q147L of S. cerevisiae full length Smc2), pGAL1 BRN1-His12-HA3 (C-terminal 12 × HIS-3 × HA-tag of S. cerevisiae full length Brn1), TRP1 | Terekawa et al., 2017 | N/A |
Plasmid 2micron pGAL1 YCG1 (S. cerevisiae full length Ycg1), pGAL10 YCS4 (S. cerevisiae full length Ycs4), URA3 | Terekawa et al., 2017 | N/A |
Plasmid 2micron pGAL7 SMC4-StrepII3 (C-terminal 3 × StrepII-tag of S. cerevisiae full length Smc4), pGAL10 SMC2 (S. cerevisiae full length Smc2), pGAL1 BRN1-His12-HA3 (C-terminal 12 × HIS-3 × HA-tag with mutated residues K409D, R411D, K414D, K451D, K452D, K454D, K456D, K457D of S. cerevisiae full length Brn1), TRP1 | This work | N/A |
Plasmid 2micron pGAL7 SMC4-StrepII3 (C-terminal 3 × StrepII-tag of S. cerevisiae full length Smc4), pGAL10 SMC2 (S. cerevisiae full length Smc2), pGAL1 BRN1-His12-HA3 (C-terminal 12 × HIS-3 × HA-tag with mutated residues M391D, F394D, W402D, W408D of S. cerevisiae full length Brn1), TRP1 | This work | N/A |
Plasmid 2micron pGAL10 YCS4 (S. cerevisiae full length Ycs4), URA3 | This work | N/A |
Plasmid pETMCN-Brn1384-529-Ycg16-932, Δ499-555 (N-terminal 6 × HIS-tag-HRV3C-cleavage-site, residues 384-529 of S. cerevisiae Brn1 and residues 6-932 with deleted residues 499-555 of untagged S. cerevisiae Ycg1) | This work | N/A |
Plasmid pETMCN-Brn1384-529, short kleisin loop-Ycg16-932, Δ499-555 (N-terminal 6 × HIS-tag-HRV3C-cleavage-site, residues 384-529 with deleted residues 418-444 of S. cerevisiae Brn1 and residues 6-932 with deleted residues 499-555 of untagged S. cerevisiae Ycg1) | This work | N/A |
Plasmid pETMCN-Cnd2416-544-Cnd31-823, Δ439-473 (N-terminal 6 × HIS-tag-HRV3C-cleavage-site, residues 416-544 of S. pombe Cnd2 and residues 1-823 with deleted residues 439-473 of untagged S. pombe Cnd3) | This work | N/A |
Plasmid pC1-NCAPH (N-terminal Flag-EGFP-tag of H. sapiens NCAPH) | Piazza et al., 2014 | N/A |
Plasmid pC1-NCAPH2 (N-terminal Flag-EGFP-tag of H. sapiens NCAPH2) | Piazza et al., 2014 | N/A |
Plasmid pC1-NCAPHBC1/2 (N-terminal Flag-EGFP-tag with mutations R446D, R448D, R450D, R451D, K452D, K462D, K463D, K464D, K467D, K468D of H. sapiens NCAPH) | This work | N/A |
Plasmid pC1-NCAPH ΦA (N-terminal Flag-EGFP-tag with mutations Y428A, F431A, W439A, W445A of H. sapiens NCAPH) | This work | N/A |
Plasmid pC1-NCAPH ΦD (N-terminal Flag-EGFP-tag with mutations Y428D, F431D, W439D, W445D of H. sapiens NCAPH) | This work | N/A |
Plasmid pC1-NCAPH S432A (N-terminal Flag-EGFP-tag with mutations S432A of H. sapiens NCAPH) | This work | N/A |
Plasmid pC1-NCAPH2BC1/2 (N-terminal Flag-EGFP-tag with mutations K329D, K332D, K333D, R335D, K350D, R351D, K352D, R353D, K354D of H. sapiens NCAPH2) | This work | N/A |
Software and Algorithms | ||
X-ray Detector Software (XDS) | Kabsch, 2010 | http://xds.mpimf-heidelberg.mpg.de/ |
SHELX | Sheldrick, 2008 | http://shelx.uni-ac.gwdg.de/SHELX/ |
Phenix suite | Adams et al., 2010 | https://www.phenix-online.org/ |
CCP4 suite | Winn et al., 2011 | http://www.ccp4.ac.uk/ |
COOT v0.8.2 | Emsley et al., 2010 | https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/ |
PyMOL | Schrödinger, LLC | https://www.pymol.org/ |
ConSurf | Ashkenazy et al., 2016 | http://bental.tau.ac.il/new_ConSurfDB/ |
APBS | Baker et al., 2001 | http://www.poissonboltzmann.org/ |
PISA | Krissinel and Henrick, 2007 | http://www.ccp4.ac.uk/MG/ccp4mg_help/pisa.html |