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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-20840 | |||||||||
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Title | FACT_subnucleosome complex 1 | |||||||||
![]() | Sharpened map | |||||||||
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![]() | assembly / ![]() ![]() | |||||||||
Function / homology | ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() Similarity search - Function | |||||||||
Biological species | ![]() ![]() ![]() ![]() ![]() | |||||||||
Method | ![]() ![]() | |||||||||
![]() | Zhou K / Tan YZ | |||||||||
Funding support | ![]()
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![]() | ![]() Title: FACT caught in the act of manipulating the nucleosome. Authors: Yang Liu / Keda Zhou / Naifu Zhang / Hui Wei / Yong Zi Tan / Zhening Zhang / Bridget Carragher / Clinton S Potter / Sheena D'Arcy / Karolin Luger / ![]() Abstract: The organization of genomic DNA into nucleosomes profoundly affects all DNA-related processes in eukaryotes. The histone chaperone known as 'facilitates chromatin transcription' (FACT) (consisting of ...The organization of genomic DNA into nucleosomes profoundly affects all DNA-related processes in eukaryotes. The histone chaperone known as 'facilitates chromatin transcription' (FACT) (consisting of subunits SPT16 and SSRP1) promotes both disassembly and reassembly of nucleosomes during gene transcription, DNA replication and DNA repair. However, the mechanism by which FACT causes these opposing outcomes is unknown. Here we report two cryo-electron-microscopic structures of human FACT in complex with partially assembled subnucleosomes, with supporting biochemical and hydrogen-deuterium exchange data. We find that FACT is engaged in extensive interactions with nucleosomal DNA and all histone variants. The large DNA-binding surface on FACT appears to be protected by the carboxy-terminal domains of both of its subunits, and this inhibition is released by interaction with H2A-H2B, allowing FACT-H2A-H2B to dock onto a complex containing DNA and histones H3 and H4 (ref. ). SPT16 binds nucleosomal DNA and tethers H2A-H2B through its carboxy-terminal domain by acting as a placeholder for DNA. SSRP1 also contributes to DNA binding, and can assume two conformations, depending on whether a second H2A-H2B dimer is present. Our data suggest a compelling mechanism for how FACT maintains chromatin integrity during polymerase passage, by facilitating removal of the H2A-H2B dimer, stabilizing intermediate subnucleosomal states and promoting nucleosome reassembly. Our findings reconcile discrepancies regarding the many roles of FACT and underscore the dynamic interactions between histone chaperones and nucleosomes. | |||||||||
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 | ![]() | 28.6 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 26.8 KB 26.8 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 7.7 KB | Display | ![]() |
Images | ![]() | 150.7 KB | ||
Masks | ![]() | 30.5 MB | ![]() | |
Filedesc metadata | ![]() | 7.7 KB | ||
Others | ![]() ![]() ![]() | 9.6 MB 28.3 MB 28.3 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6upkMC ![]() 6uplC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | |
EM raw data | ![]() Data size: 2.2 TB Data #1: Unaligned multi-frame micrographs [micrographs - multiframe] Data #2: Aligned and dose-weighted micrographs [micrographs - single frame] Data #3: Final Particle Stacks for Class 1 and 2 with Euler Angles and Shifts [picked particles - single frame - processed]) |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Sharpened map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.40301 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
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Projections & Slices |
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Density Histograms |
-Additional map: unfiltered map
File | emd_20840_additional.map | ||||||||||||
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Annotation | unfiltered map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_20840_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_20840_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
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Sample components
-Entire : FACT_subNucleosome_complex_class1
Entire | Name: FACT_subNucleosome_complex_class1 |
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Components |
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-Supramolecule #1: FACT_subNucleosome_complex_class1
Supramolecule | Name: FACT_subNucleosome_complex_class1 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 320 KDa |
-Macromolecule #1: Histone H3.1
Macromolecule | Name: Histone H3.1 / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 15.437167 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MARTKQTARK STGGKAPRKQ LATKAARKSA PATGGVKKPH RYRPGTVALR EIRRYQKSTE LLIRKLPFQR LVREIAQDFK TDLRFQSSA VMALQEACEA YLVGLFEDTN LCAIHAKRVT IMPKDIQLAR RIRGERA UniProtKB: ![]() |
-Macromolecule #2: Histone H4
Macromolecule | Name: Histone H4 / type: protein_or_peptide / ID: 2 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 11.394426 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MSGRGKGGKG LGKGGAKRHR KVLRDNIQGI TKPAIRRLAR RGGVKRISGL IYEETRGVLK VFLENVIRDA VTYTEHAKRK TVTAMDVVY ALKRQGRTLY GFGG UniProtKB: ![]() |
-Macromolecule #3: Histone H2A
Macromolecule | Name: Histone H2A / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 14.135523 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MSGRGKQGGK ARAKAKSRSS RAGLQFPVGR VHRLLRKGNY AERVGAGAPV YLAAVLEYLT AEILELAGNA ARDNKKTRII PRHLQLAIR NDEELNKLLG RVTIAQGGVL PNIQAVLLPK KTESHHKAKG K UniProtKB: Histone H2A type 1-C |
-Macromolecule #4: Histone H2B
Macromolecule | Name: Histone H2B / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 13.937213 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MPEPAKSAPA PKKGSKKAVT KAQKKDGKKR KRSRKESYSV YVYKVLKQVH PDTGISSKAM GIMNSFVNDI FERIAGEASR LAHYNKRST ITSREIQTAV RLLLPGELAK HAVSEGTKAV TKYTSSK UniProtKB: Histone H2B type 1-C/E/F/G/I |
-Macromolecule #5: FACT complex subunit SPT16
Macromolecule | Name: FACT complex subunit SPT16 / type: protein_or_peptide / ID: 5 Details: Amino acids 640-651 cannot be identified due to the resolution limitations. To show the connectivity of the electron density, poly-(UNK) was placed. This was also done for the end of this ...Details: Amino acids 640-651 cannot be identified due to the resolution limitations. To show the connectivity of the electron density, poly-(UNK) was placed. This was also done for the end of this chain (926-967). The full sequence for the experiment is: MHHHHHHAVTLDKDAYYRRVKRLYSNWRKGEDEYANVDAIVVSVGVDEEIVYAKSTALQTWLFGYELTDTIMVFCDDKII FMASKKKVEFLKQIANTKGNENANGAPAITLLIREKNESNKSSFDKMIEAIKESKNGKKIGVFSKDKFPGEFMKSWNDCL NKEGFDKIDISAVVAYTIAVKEDGELNLMKKAASITSEVFNKFFKERVMEIVDADEKVRHSKLAESVEKAIEEKKYLAGA DPSTVEMCYPPIIQSGGNYNLKFSVVSDKNHMHFGAITCAMGIRFKSYCSNLVRTLMVDPSQEVQENYNFLLQLQEELLK ELRHGVKICDVYNAVMDVVKKQKPELLNKITKNLGFGMGIEFREGSLVINSKNQYKLKKGMVFSINLGFSDLTNKEGKKP EEKTYALFIGDTVLVDEDGPATVLTSVKKKVKNVGIFLKNEDEEEEEEEKDEAEDLLGRGSRAALLTERTRNEMTAEEKR RAHQKELAAQLNEEAKRRLTEQKGEQQIQKARKSNVSYKNPSLMPKEPHIREMKIYIDKKYETVIMPVFGIATPFHIATI KNISMSVEGDYTYLRINFYCPGSALGRNEGNIFPNPEATFVKEITYRASNIKAPGEQTVPALNLQNAFRIIKEVQKRYKT REAEEKEKEGIVKQDSLVINLNRSNPKLKDLYIRPNIAQKRMQGSLEAHVNGFRFTSVRGDKVDILYNNIKHALFQPCDG EMIIVLHFHLKNAIMFGKKRHTDVQFYTEVGEITTDLGKHQHMHDRDDLYAEQMEREMRHKLKTAFKNFIEKVEALTKEE LEFEVPFRDLGFNGAPYRSTCLLQPTSSALVNATEWPPFVVTLDEVELIHFERVQFHLKNFDMVIVYKDYSKKVTMINAI PVASLDPIKEWLNSCDLKYTEGVQSLNWTKIMKTIVDDPEGFFEQGGWSFLEPEGEGSDAEEGDSESEIEDETFNPSEDD YEEEEEDSDEDYSSEAEESDYSKESLGSEEESGKDWDELEEEARKADRESRYEEEEEQSRSMSRKRKASVHSSGRGSNRG SRHSSAPPKKKRK Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 109.574867 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MHHHHHHAVT LDKDAYYRRV KRLYSNWRKG EDEYANVDAI VVSVGVDEEI VYAKSTALQT WLFGYELTDT IMVFCDDKII FMASKKKVE FLKQIANTKG NENANGAPAI TLLIREKNES NKSSFDKMIE AIKESKNGKK IGVFSKDKFP GEFMKSWNDC L NKEGFDKI ...String: MHHHHHHAVT LDKDAYYRRV KRLYSNWRKG EDEYANVDAI VVSVGVDEEI VYAKSTALQT WLFGYELTDT IMVFCDDKII FMASKKKVE FLKQIANTKG NENANGAPAI TLLIREKNES NKSSFDKMIE AIKESKNGKK IGVFSKDKFP GEFMKSWNDC L NKEGFDKI DISAVVAYTI AVKEDGELNL MKKAASITSE VFNKFFKERV MEIVDADEKV RHSKLAESVE KAIEEKKYLA GA DPSTVEM CYPPIIQSGG NYNLKFSVVS DKNHMHFGAI TCAMGIRFKS YCSNLVRTLM VDPSQEVQEN YNFLLQLQEE LLK ELRHGV KICDVYNAVM DVVKKQKPEL LNKITKNLGF GMGIEFREGS LVINSKNQYK LKKGMVFSIN LGFSDLTNKE GKKP EEKTY ALFIGDTVLV DEDGPATVLT SVKKKVKNVG IFLKNEDEEE EEEEKDEAED LLGRGSRAAL LTERTRNEMT AEEKR RAHQ KELAAQLNEE AKRRLTEQKG EQQIQKARKS NVSYKNPSLM PKEPHIREMK IYIDKKYETV IMPVFGIATP FHIATI KNI SMSVEGDYTY LRINFYCPGS ALGRNEGNIF PNPEATFVKE ITYRASNIKA PGEQTVPALN LQNAFRIIKE VQKRYK (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)V KQDSLVINLN RSNPKLKDL YIRPNIAQKR MQGSLEAHVN GFRFTSVRGD KVDILYNNIK HALFQPCDGE MIIVLHFHLK NAIMFGKKRH TDVQFYTEVG EITTDLGKH QHMHDRDDLY AEQMEREMRH KLKTAFKNFI EKVEALTKEE LEFEVPFRDL GFNGAPYRST CLLQPTSSAL V NATEWPPF VVTLDEVELI HFERVQFHLK NFDMVIVYKD YSKKVTMINA IPVASLDPIK EWLNSCDLKY TEGVQSLNWT KI MKTIVDD PEGFFEQGGW SFL(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) |
-Macromolecule #6: FACT complex subunit SSRP1
Macromolecule | Name: FACT complex subunit SSRP1 / type: protein_or_peptide / ID: 6 Details: The poly-(UNK) was placed to show the electron density between 171-198. The full sequence for the experiment is: ...Details: The poly-(UNK) was placed to show the electron density between 171-198. The full sequence for the experiment is: MAETLEFNDVYQEVKGSMNDGRLRLSRQGIIFKNSKTGKVDNIQAGELTEGIWRRVALGHGLKLLTKNGHVYKYDGFRES EFEKLSDFFKTHYRLELMEKDLCVKGWNWGTVKFGGQLLSFDIGDQPVFEIPLSNVSQCTTGKNEVTLEFHQNDDAEVSL MEVRFYVPPTQEDGVDPVEAFAQNVLSKADVIQATGDAICIFRELQCLTPRGRYDIRIYPTFLHLHGKTFDYKIPYTTVL RLFLLPHKDQRQMFFVISLDPPIKQGQTRYHFLILLFSKDEDISLTLNMNEEEVEKRFEGRLTKNMSGSLYEMVSRVMKA LVNRKITVPGNFQGHSGAQCITCSYKASSGLLYPLERGFIYVHKPPVHIRFDEISFVNFARGTTTTRSFDFEIETKQGTQ YTFSSIEREEYGKLFDFVNAKKLNIKNRGLKEGMNPSYDEYADSDEDQHDAYLERMKEEGKIREENANDSSDDSGEETDE SFNPGEEEEDVAEEFDSNASASSSSNEGDSDRDEKKRKQLKKAKMAKDRKSRKKPVEVKKGKDPNAPKRPMSAYMLWLNA SREKIKSDHPGISITDLSKKAGEIWKGMSKEKKEEWDRKAEDARRDYEKAMKEYEGGRGESSKRDKSKKKKKVKVKMEKK STPSRGSSSKSSSRQLSESFKSKEFVSSDESSSGENKSKKKRRRSEDSEEEELASTPPSSEDSASGSDE Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 73.361172 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MAETLEFNDV YQEVKGSMND GRLRLSRQGI IFKNSKTGKV DNIQAGELTE GIWRRVALGH GLKLLTKNGH VYKYDGFRES EFEKLSDFF KTHYRLELME KDLCVKGWNW GTVKFGGQLL SFDIGDQPVF EIPLSNVSQC TTGKNEVTLE FHQNDDAEVS L MEVRFYVP ...String: MAETLEFNDV YQEVKGSMND GRLRLSRQGI IFKNSKTGKV DNIQAGELTE GIWRRVALGH GLKLLTKNGH VYKYDGFRES EFEKLSDFF KTHYRLELME KDLCVKGWNW GTVKFGGQLL SFDIGDQPVF EIPLSNVSQC TTGKNEVTLE FHQNDDAEVS L MEVRFYVP PTQ(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)D AICIFRELQC LTPRGRYDI RIYPTFLHLH GKTFDYKIPY TTVLRLFLLP HKDQRQMFFV ISLDPPIKQG QTRYHFLILL FSKDEDISLT LNMNEEEVEK RFEGRLTKN MSGSLYEMVS RVMKALVNRK ITVPGNFQGH SGAQCITCSY KASSGLLYPL ERGFIYVHKP PVHIRFDEIS F VNFARGTT TTRSFDFEIE TKQGTQYTFS SIEREEYGKL FDFVNAKKLN IKNRGLKEGM NPSYDEYADS DEDQHDAYLE RM KEEGKIR EENANDSSDD SGEETDESFN PGEEEEDVAE EFDSNASASS SSNEGDSDRD EKKRKQLKKA KMAKDRKSRK KPV EVKKGK DPNAPKRPMS AYMLWLNASR EKIKSDHPGI SITDLSKKAG EIWKGMSKEK KEEWDRKAED ARRDYEKAMK EYEG GRGES SKRDKSKKKK KVKVKMEKK |
-Macromolecule #7: DNA (79-mer)
Macromolecule | Name: DNA (79-mer) / type: dna / ID: 7 / Number of copies: 1 / Classification: DNA |
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Source (natural) | Organism: ![]() ![]() ![]() |
Molecular weight | Theoretical: 24.144422 KDa |
Sequence | String: (DT)(DC)(DG)(DT)(DA)(DG)(DA)(DC)(DA)(DG) (DC)(DT)(DC)(DT)(DA)(DG)(DC)(DA)(DC)(DC) (DG)(DC)(DT)(DT)(DA)(DA)(DA)(DC)(DG) (DC)(DA)(DC)(DG)(DT)(DA)(DC)(DG)(DC)(DG) (DC) (DT)(DG)(DT)(DC)(DC)(DC) ...String: (DT)(DC)(DG)(DT)(DA)(DG)(DA)(DC)(DA)(DG) (DC)(DT)(DC)(DT)(DA)(DG)(DC)(DA)(DC)(DC) (DG)(DC)(DT)(DT)(DA)(DA)(DA)(DC)(DG) (DC)(DA)(DC)(DG)(DT)(DA)(DC)(DG)(DC)(DG) (DC) (DT)(DG)(DT)(DC)(DC)(DC)(DC)(DC) (DG)(DC)(DG)(DT)(DT)(DT)(DT)(DA)(DA)(DC) (DC)(DG) (DC)(DC)(DA)(DA)(DG)(DG)(DG) (DG)(DA)(DT)(DT)(DA)(DC)(DT)(DC)(DC)(DC) (DT)(DA) |
-Macromolecule #8: DNA (79-mer)
Macromolecule | Name: DNA (79-mer) / type: dna / ID: 8 / Number of copies: 1 / Classification: DNA |
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Source (natural) | Organism: ![]() ![]() ![]() |
Molecular weight | Theoretical: 24.58468 KDa |
Sequence | String: (DT)(DA)(DG)(DG)(DG)(DA)(DG)(DT)(DA)(DA) (DT)(DC)(DC)(DC)(DC)(DT)(DT)(DG)(DG)(DC) (DG)(DG)(DT)(DT)(DA)(DA)(DA)(DA)(DC) (DG)(DC)(DG)(DG)(DG)(DG)(DG)(DA)(DC)(DA) (DG) (DC)(DG)(DC)(DG)(DT)(DA) ...String: (DT)(DA)(DG)(DG)(DG)(DA)(DG)(DT)(DA)(DA) (DT)(DC)(DC)(DC)(DC)(DT)(DT)(DG)(DG)(DC) (DG)(DG)(DT)(DT)(DA)(DA)(DA)(DA)(DC) (DG)(DC)(DG)(DG)(DG)(DG)(DG)(DA)(DC)(DA) (DG) (DC)(DG)(DC)(DG)(DT)(DA)(DC)(DG) (DT)(DG)(DC)(DG)(DT)(DT)(DT)(DA)(DA)(DG) (DC)(DG) (DG)(DT)(DG)(DC)(DT)(DA)(DG) (DA)(DG)(DC)(DT)(DG)(DT)(DC)(DT)(DA)(DC) (DG)(DA) |
-Experimental details
-Structure determination
Method | ![]() |
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Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD![]() |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 7.8 e/Å2 |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |