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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-4029 | |||||||||
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Title | An engineered human cohesin complex bound to Pds5B | |||||||||
![]() | Pds5B-bound bonsai cohesin tetramer | |||||||||
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Biological species | ![]() | |||||||||
Method | single particle reconstruction / negative staining / Resolution: 31.0 Å | |||||||||
![]() | Hons MT / Huis in 't Veld PJ / Stark H / Peters J-M | |||||||||
![]() | ![]() Title: Topology and structure of an engineered human cohesin complex bound to Pds5B. Authors: Michael T Hons / Pim J Huis In 't Veld / Jan Kaesler / Pascaline Rombaut / Alexander Schleiffer / Franz Herzog / Holger Stark / Jan-Michael Peters / ![]() ![]() Abstract: The cohesin subunits Smc1, Smc3 and Scc1 form large tripartite rings which mediate sister chromatid cohesion and chromatin structure. These are thought to entrap DNA with the help of the associated ...The cohesin subunits Smc1, Smc3 and Scc1 form large tripartite rings which mediate sister chromatid cohesion and chromatin structure. These are thought to entrap DNA with the help of the associated proteins SA1/2 and Pds5A/B. Structural information is available for parts of cohesin, but analyses of entire cohesin complexes are limited by their flexibility. Here we generated a more rigid 'bonsai' cohesin by truncating the coiled coils of Smc1 and Smc3 and used single-particle electron microscopy, chemical crosslinking-mass spectrometry and in silico modelling to generate three-dimensional models of cohesin bound to Pds5B. The HEAT-repeat protein Pds5B forms a curved structure around the nucleotide-binding domains of Smc1 and Smc3 and bridges the Smc3-Scc1 and SA1-Scc1 interfaces. These results indicate that Pds5B forms an integral part of the cohesin ring by contacting all other cohesin subunits, a property that may reflect the complex role of Pds5 proteins in controlling cohesin-DNA interactions. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 1.5 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 18 KB 18 KB | Display Display | ![]() |
Images | ![]() | 73.4 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 189.5 KB | Display | ![]() |
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Full document | ![]() | 188.6 KB | Display | |
Data in XML | ![]() | 5.6 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | Pds5B-bound bonsai cohesin tetramer | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.51 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Petameric bonsai cohesin complex including Pds5B
Entire | Name: Petameric bonsai cohesin complex including Pds5B |
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Components |
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-Supramolecule #1: Petameric bonsai cohesin complex including Pds5B
Supramolecule | Name: Petameric bonsai cohesin complex including Pds5B / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() ![]() |
Molecular weight | Theoretical: 529 KDa |
-Macromolecule #1: SMC1_truncated
Macromolecule | Name: SMC1_truncated / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Sequence | String: MGFLKLIEIE NFKSYKGRQI IGPFQRFTAI IGPNGSGKSN LMDAISFVLG EKTSNLRVKT LRDLIHGAPV GKPAANRAFV SMVYSEEGAE DRTFARVIVG GSSEYKINNK VVQLHEYSEE LEKLGILIKA RNFLVFQGAV ESIAMKNPKE RTALFEEISR SGELAQEYDK ...String: MGFLKLIEIE NFKSYKGRQI IGPFQRFTAI IGPNGSGKSN LMDAISFVLG EKTSNLRVKT LRDLIHGAPV GKPAANRAFV SMVYSEEGAE DRTFARVIVG GSSEYKINNK VVQLHEYSEE LEKLGILIKA RNFLVFQGAV ESIAMKNPKE RTALFEEISR SGELAQEYDK RKKEMVKAEE DTQFNYHRKK NIAAERKEAK PGRKAEIMES IKRLYPGSVY GRLIDLCQPT QKKYQIAVTK VLGKNMDAII VDSEKTGRDC IQYIKEQRGE PETFLPLDYL EVKPTDEKLR ELKGAKLVID VIRYEPPHIK KALQYACGNA LVCDNVEDAR RIAFGGHQRH KTVALDGTLF QKSGVISGGA SDLKAKARRW DEKAVDKLKS RLIEIDYGDL CEDLKDAQAE EEIKQEMNTL QQKLNEQQSV LQRIAAPNMK AMEKLESVRD KFQETSDEFE AARKRAKKAK QAFEQIKKER FDRFNACFES VATNIDEIYK ALSRNSSAQA FLGPENPEEP YLDGINYNCV APGKRFRPMD NLSGGEKTVA ALALLFAIHS YKPAPFFVLD EIDAALDNTN IGKVANYIKE QSTCNFQAIV ISLKEEFYTK AESLIGVYPE QGDCVISKVL TFDLTKYPDA NPNPNEQ |
-Macromolecule #2: SMC3_truncated
Macromolecule | Name: SMC3_truncated / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MYIKQVIIQG FRSYRDQTIV DPFSSKHNVI VGRNGSGKSN FFYAIQFVLS DEFSHLRPEQ RLALLHEGTG PRVISAFVEI IFDNSDNRLP IDKEEVSLRR VIGAKKDQYF LDKKMVTKND VMNLLESAGF SRSNPYYIVK QGKINQMATA PDSQRLKLLR EVAGTRVYDE ...String: MYIKQVIIQG FRSYRDQTIV DPFSSKHNVI VGRNGSGKSN FFYAIQFVLS DEFSHLRPEQ RLALLHEGTG PRVISAFVEI IFDNSDNRLP IDKEEVSLRR VIGAKKDQYF LDKKMVTKND VMNLLESAGF SRSNPYYIVK QGKINQMATA PDSQRLKLLR EVAGTRVYDE RKEESISLMK ETEGKREKIN ELLKYIEERL HTLEGTKKQQ LLRAATGKAI LNGIDSINKV LDHFRRKGIN QHVQNGYHGI VMNNFECEPA FYTCVEVTAG NRLFYHIVDS DEVSTKILME FNKMNLPGEV TFLPLNKLDV RDTAYPETND AIPMISKLRY NPRFDKAFKH VFGKTLICRS MEVSTQLARA FTMDCITLEG DQVSHRGALT GGYYDTRKSR LELQKDVRKA ARYQTLSLKQ LFRKLEQCNT ELKKYSHVNK KALDQFVNFS EQKEKLIKRQ EELDRGYKSI MELMNVLELR KYEAIQLTFK QVSKNFSEVF QKLVPGGKAT LVMKKGDVEG SQSQDEGEGS GESERGSGSQ SSVPSVDQFT GVGIRVSFTG KQGEMREMQQ LSGGQKSLVA LALIFAIQKC DPAPFYLFDE IDQALDAQHR KAVSDMIMEL AVHAQFITTT FRPELLESAD KFYGVKFRNK VSHIDVITAE MAKDFVEDDT THGDYKDDDD K |
-Macromolecule #3: SCC1
Macromolecule | Name: SCC1 / type: protein_or_peptide / ID: 3 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MFYAHFVLSK RGPLAKIWLA AHWDKKLTKA HVFECNLESS VESIISPKVK MALRTSGHLL LGVVRIYHRK AKYLLADCNE AFIKIKMAFR PGVVDLPEEN REAAYNAITL PEEFHDFDQP LPDLDDIDVA QQFSLNQSRV EEITMREEVG NISILQENDF GDFGMDDREI ...String: MFYAHFVLSK RGPLAKIWLA AHWDKKLTKA HVFECNLESS VESIISPKVK MALRTSGHLL LGVVRIYHRK AKYLLADCNE AFIKIKMAFR PGVVDLPEEN REAAYNAITL PEEFHDFDQP LPDLDDIDVA QQFSLNQSRV EEITMREEVG NISILQENDF GDFGMDDREI MREGSAFEDD DMLVSTTTSN LLLESEQSTS NLNEKINHLE YEDQYKDDNF GEGNDGGILD DKLISNNDGG IFDDPPALSE AGVMLPEQPA HDDMDEDDNV SMGGPDSPDS VDPVEPMPTM TDQTTLVPNE EEAFALEPID ITVKETKAKR KRKLIVDSVK ELDSKTIRAQ LSDYSDIVTT LDLAPPTKKL MMWKETGGVE KLFSLPAQPL WNNRLLKLFT RCLTPLVPED LRKRRKGGEA DNLDEFLKEF ENPEVPREDQ QQQHQQRDVI DEPIIEEPSR LQESVMEASR TNIDESAMPP PPPQGVKRKA GQIDPEPVMP PQQVEQMEIP PVELPPEEPP NICQLIPELE LLPEKEKEKE KEKEDDEEEE DEDASGGDQD QEERRWNKRT QQMLHGLQRA LAKTGAESIS LLELCRNTNR KQAAAKFYSF LVLKKQQAIE LTQEEPYSDI IATPGPRFHI I |
-Macromolecule #4: SA1
Macromolecule | Name: SA1 / type: protein_or_peptide / ID: 4 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MHHHHHHHHH HAGGAITSEL PVLQDSTNET TAHSDAGSEL EETEVKGKRK RGRPGRPPST NKKPRKSPGE KSRIEAGIRG AGRGRANGHP QQNGEGEPVT LFEVVKLGKS AMQSVVDDWI ESYKQDRDIA LLDLINFFIQ CSGCRGTVRI EMFRNMQNAE IIRKMTEEFD ...String: MHHHHHHHHH HAGGAITSEL PVLQDSTNET TAHSDAGSEL EETEVKGKRK RGRPGRPPST NKKPRKSPGE KSRIEAGIRG AGRGRANGHP QQNGEGEPVT LFEVVKLGKS AMQSVVDDWI ESYKQDRDIA LLDLINFFIQ CSGCRGTVRI EMFRNMQNAE IIRKMTEEFD EDSGDYPLTM PGPQWKKFRS NFCEFIGVLI RQCQYSIIYD EYMMDTVISL LTGLSDSQVR AFRHTSTLAA MKLMTALVNV ALNLSIHQDN TQRQYEAERN KMIGKRANER LELLLQKRKE LQENQDEIEN MMNSIFKGIF VHRYRDAIAE IRAICIEEIG VWMKMYSDAF LNDSYLKYVG WTLHDRQGEV RLKCLKALQS LYTNRELFPK LELFTNRFKD RIVSMTLDKE YDVAVEAIRL VTLILHGSEE ALSNEDCENV YHLVYSAHRP VAVAAGEFLH KKLFSRHDPQ AEEALAKRRG RNSPNGNLIR MLVLFFLESE LHEHAAYLVD SLWESSQELL KDWECMTELL LEEPVQGEEA MSDRQESALI ELMVCTIRQA AEAHPPVGRG TGKRVLTAKE RKTQIDDRNK LTEHFIITLP MLLSKYSADA EKVANLLQIP QYFDLEIYST GRMEKHLDAL LKQIKFVVEK HVESDVLEAC SKTYSILCSE EYTIQNRVDI ARSQLIDEFV DRFNHSVEDL LQEGEEADDD DIYNVLSTLK RLTSFHNAHD LTKWDLFGNC YRLLKTGIEH GAMPEQIVVQ ALQCSHYSIL WQLVKITDGS PSKEDLLVLR KTVKSFLAVC QQCLSNVNTP VKEQAFMLLC DLLMIFSHQL MTGGREGLQP LVFNPDTGLQ SELLSFVMDH VFIDQDEENQ SMEGDEEDEA NKIEALHKRR NLLAAFSKLI IYDIVDMHAA ADIFKHYMKY YNDYGDIIKE TLSKTRQIDK IQCAKTLILS LQQLFNELVQ EQGPNLDRTS AHVSGIKELA RRFALTFGLD QIKTREAVAT LHKDGIEFAF KYQNQKGQEY PPPNLAFLEV LSEFSSKLLR QDKKTVHSYL EKFLTEQMME RREDVWLPLI SYRNSLVTGG EDDRMSVNSG SSSSKTSSVR NKKGRPPLHK KRVEDESLDN TWLNRTDTMI QTPGPLPAPQ LTSTVLRENS RPMGDQIQEP ESEHGSEPDF LHNPQMQISW LGQPKLEDLN RKDRTGMNYM KVRTGVRHAV RGLMEEDAEP IFEDVMMSSR SQLEDMNEEF EDTMVIDLPP SRNRRERAEL RPDFFDSAAI IEDDSGFGMP MF |
-Macromolecule #5: PDS5B
Macromolecule | Name: PDS5B / type: protein_or_peptide / ID: 5 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MHHHHHHHHH HAGGAAHSKT RTNDGKITYP PGVKEISDKI SKEEMVRRLK MVVKTFMDMD QDSEEEKELY LNLALHLASD FFLKHPDKDV RLLVACCLAD IFRIYAPEAP YTSPDKLKDI FMFITRQLKG LEDTKSPQFN RYFYLLENIA WVKSYNICFE LEDSNEIFTQ ...String: MHHHHHHHHH HAGGAAHSKT RTNDGKITYP PGVKEISDKI SKEEMVRRLK MVVKTFMDMD QDSEEEKELY LNLALHLASD FFLKHPDKDV RLLVACCLAD IFRIYAPEAP YTSPDKLKDI FMFITRQLKG LEDTKSPQFN RYFYLLENIA WVKSYNICFE LEDSNEIFTQ LYRTLFSVIN NGHNQKVHMH MVDLMSSIIC EGDTVSQELL DTVLVNLVPA HKNLNKQAYD LAKALLKRTA QAIEPYITNF FNQVLMLGKT SISDLSEHVF DLILELYNID SHLLLSVLPQ LEFKLKSNDN EERLQVVKLL AKMFGAKDSE LASQNKPLWQ CYLGRFNDIH VPIRLECVKF ASHCLMNHPD LAKDLTEYLK VRSHDPEEAI RHDVIVSIVT AAKKDILLVN DHLLNFVRER TLDKRWRVRK EAMMGLAQIY KKYALQSAAG KDAAKQIAWI KDKLLHIYYQ NSIDDRLLVE RIFAQYMVPH NLETTERMKC LYYLYATLDL NAVKALNEMW KCQNLLRHQV KDLLDLIKQP KTDASVKAIF SKVMVITRNL PDPGKAQDFM KKFTQVLEDD EKIRKQLEVL VSPTCSCKQA EGCVREITKK LGNPKQPTNP FLEMIKFLLE RIAPVHIDTE SISALIKQVN KSIDGTADDE DEGVPTDQAI RAGLELLKVL SFTHPISFHS AETFESLLAC LKMDDEKVAE AALQIFKNTG SKIEEDFPHI RSALLPVLHH KSKKGPPRQA KYAIHCIHAI FSSKETQFAQ IFEPLHKSLD PSNLEHLITP LVTIGHIALL APDQFAAPLK SLVATFIVKD LLMNDRLPGK KTTKLWVPDE EVSPETMVKI QAIKMMVRWL LGMKNNHSKS GTSTLRLLTT ILHSDGDLTE QGKISKPDMS RLRLAAGSAI VKLAQEPCYH EIITLEQYQL CALAINDECY QVRQVFAQKL HKGLSRLRLP LEYMAICALC AKDPVKERRA HARQCLVKNI NVRREYLKQH AAVSEKLLSL LPEYVVPYTI HLLAHDPDYV KVQDIEQLKD VKECLWFVLE ILMAKNENNS HAFIRKMVEN IKQTKDAQGP DDAKMNEKLY TVCDVAMNII MSKSTTYSLE SPKDPVLPAR FFTQPDKNFS NTKNYLPPEM KSFFTPGKPK TTNVLGAVNK PLSSAGKQSQ TKSSRMETVS NASSSSNPSS PGRIKGRLDS SEMDHSENED YTMSSPLPGK KSDKRDDSDL VRSELEKPRG RKKTPVTEQE EKLGMDDLTK LVQEQKPKGS QRSRKRGHTA SESDEQQWPE EKRLKEDILE NEDEQNSPPK KGKRGRPPKP LGGGTPKEEP TMKTSKKGSK KKSGPPAPEE EEEEERQSGN TEQKSKSKQH RVSRRAQQRA ESPESSAIES TQSTPQKGRG RPSKTPSPSQ PKKNVRVGRS KQAATKENDS SEEVDVFQGS SPVDDIPQEE TEEEEVSTVN VRRRSAKRER R |
-Experimental details
-Structure determination
Method | negative staining |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.5 Component:
Details: Continuous 15-40% (v/v) glycerol gradients with a 10% glycerol cushion (0.1 mL): 25 mM Hepes pH 7.5, 150 mM NaCl, 2mM MgCl2 0.183 mM ATPgammaS stabilization was accomplished by a modified ...Details: Continuous 15-40% (v/v) glycerol gradients with a 10% glycerol cushion (0.1 mL): 25 mM Hepes pH 7.5, 150 mM NaCl, 2mM MgCl2 0.183 mM ATPgammaS stabilization was accomplished by a modified GraFix protocol: 0.1% (v/v) glutaraldehyde in the 40% glycerol fraction 1 mM N-(p-Maleimidophenyl)isocyanate (PMPI) in DMSO in the 15% glycerol fraction | ||||||||||
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Staining | Type: NEGATIVE / Material: Uranyl Formate | ||||||||||
Grid | Model: Quantifoil R3.5/1 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY |
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Electron microscopy
Microscope | FEI/PHILIPS CM200FEG |
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Image recording | Film or detector model: TVIPS TEMCAM-F415 (4k x 4k) / Average electron dose: 45.0 e/Å2 |
Electron beam | Acceleration voltage: 160 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD |
Sample stage | Cooling holder cryogen: NITROGEN |