+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-14198 | |||||||||
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Title | Refined, unsharpened map of human telomerase-DNA-TPP1 complex | |||||||||
Map data | Refined, unsharpened map of human telomerase-TPP1-DNA | |||||||||
Sample |
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Keywords | Reverse transcriptase / ribonucleoprotein / telomerase / telomere / DNA BINDING PROTEIN | |||||||||
Function / homology | Function and homology information positive regulation of hair cycle / template-free RNA nucleotidyltransferase / positive regulation of transdifferentiation / TERT-RMRP complex / DNA strand elongation / RNA-directed RNA polymerase complex / positive regulation of protein localization to nucleolus / siRNA transcription / telomerase catalytic core complex / RNA-templated DNA biosynthetic process ...positive regulation of hair cycle / template-free RNA nucleotidyltransferase / positive regulation of transdifferentiation / TERT-RMRP complex / DNA strand elongation / RNA-directed RNA polymerase complex / positive regulation of protein localization to nucleolus / siRNA transcription / telomerase catalytic core complex / RNA-templated DNA biosynthetic process / : / establishment of protein localization to telomere / telomerase activity / shelterin complex / nuclear telomere cap complex / siRNA processing / telomere capping / telomerase holoenzyme complex / positive regulation of vascular associated smooth muscle cell migration / telomerase RNA binding / DNA biosynthetic process / RNA-templated transcription / telomeric DNA binding / positive regulation of stem cell proliferation / mitochondrial nucleoid / negative regulation of cellular senescence / Telomere Extension By Telomerase / telomere maintenance via telomerase / negative regulation of extrinsic apoptotic signaling pathway in absence of ligand / replicative senescence / positive regulation of Wnt signaling pathway / positive regulation of G1/S transition of mitotic cell cycle / negative regulation of endothelial cell apoptotic process / response to cadmium ion / : / positive regulation of vascular associated smooth muscle cell proliferation / telomere maintenance / mitochondrion organization / positive regulation of nitric-oxide synthase activity / positive regulation of glucose import / Formation of the beta-catenin:TCF transactivating complex / regulation of protein stability / PML body / transcription coactivator binding / positive regulation of miRNA transcription / RNA-directed DNA polymerase / structural constituent of chromatin / positive regulation of angiogenesis / RNA-directed DNA polymerase activity / nucleosome / positive regulation of protein binding / protein-folding chaperone binding / cellular response to hypoxia / negative regulation of neuron apoptotic process / chromosome, telomeric region / tRNA binding / nuclear body / nuclear speck / protein heterodimerization activity / negative regulation of gene expression / RNA-dependent RNA polymerase activity / nucleolus / protein homodimerization activity / DNA binding / RNA binding / nucleoplasm / identical protein binding / nucleus / metal ion binding / plasma membrane / cytosol Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.2 Å | |||||||||
Authors | Sekne Z / Ghanim GE / van Roon AMM / Nguyen THD | |||||||||
Funding support | United Kingdom, United States, 2 items
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Citation | Journal: Science / Year: 2022 Title: Structural basis of human telomerase recruitment by TPP1-POT1. Authors: Zala Sekne / George E Ghanim / Anne-Marie M van Roon / Thi Hoang Duong Nguyen / Abstract: Telomerase maintains genome stability by extending the 3' telomeric repeats at eukaryotic chromosome ends, thereby counterbalancing progressive loss caused by incomplete genome replication. In ...Telomerase maintains genome stability by extending the 3' telomeric repeats at eukaryotic chromosome ends, thereby counterbalancing progressive loss caused by incomplete genome replication. In mammals, telomerase recruitment to telomeres is mediated by TPP1, which assembles as a heterodimer with POT1. We report structures of DNA-bound telomerase in complex with TPP1 and with TPP1-POT1 at 3.2- and 3.9-angstrom resolution, respectively. Our structures define interactions between telomerase and TPP1-POT1 that are crucial for telomerase recruitment to telomeres. The presence of TPP1-POT1 stabilizes the DNA, revealing an unexpected path by which DNA exits the telomerase active site and a DNA anchor site on telomerase that is important for telomerase processivity. Our findings rationalize extensive prior genetic and biochemical findings and provide a framework for future mechanistic work on telomerase regulation. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_14198.map.gz | 65.3 MB | EMDB map data format | |
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Header (meta data) | emd-14198-v30.xml emd-14198.xml | 21.5 KB 21.5 KB | Display Display | EMDB header |
Images | emd_14198.png | 49.2 KB | ||
Filedesc metadata | emd-14198.cif.gz | 7.5 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-14198 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-14198 | HTTPS FTP |
-Validation report
Summary document | emd_14198_validation.pdf.gz | 475.9 KB | Display | EMDB validaton report |
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Full document | emd_14198_full_validation.pdf.gz | 475.5 KB | Display | |
Data in XML | emd_14198_validation.xml.gz | 6.2 KB | Display | |
Data in CIF | emd_14198_validation.cif.gz | 7.1 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-14198 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-14198 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_14198.map.gz / Format: CCP4 / Size: 83.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Refined, unsharpened map of human telomerase-TPP1-DNA | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.09 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : Complex of telomeric DNA-bound human telomerase with TPP1
Entire | Name: Complex of telomeric DNA-bound human telomerase with TPP1 |
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Components |
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-Supramolecule #1: Complex of telomeric DNA-bound human telomerase with TPP1
Supramolecule | Name: Complex of telomeric DNA-bound human telomerase with TPP1 type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Homo sapiens (human) |
-Macromolecule #1: Telomerase reverse transcriptase
Macromolecule | Name: Telomerase reverse transcriptase / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Recombinant expression | Organism: Homo sapiens (human) |
Sequence | String: MPRAPRCRAV RSLLRSHYRE VLPLATFVRR LGPQGWRLVQ RGDP AAFRA LVAQCLVCVP WDARPPPAAP SFRQVSCLKE LVARVLQRLC ERGAKNVLAF GFA LLDGAR GGPPEAFTTS VRSYLPNTVT DALRGSGAWG LLLRRVGDDV LVHLLARCAL FV LVAPSCA ...String: MPRAPRCRAV RSLLRSHYRE VLPLATFVRR LGPQGWRLVQ RGDP AAFRA LVAQCLVCVP WDARPPPAAP SFRQVSCLKE LVARVLQRLC ERGAKNVLAF GFA LLDGAR GGPPEAFTTS VRSYLPNTVT DALRGSGAWG LLLRRVGDDV LVHLLARCAL FV LVAPSCA YQVCGPPLYQ LGAATQARPP PHASGPRRRL GCERAWNHSV REAGVPLGLP A PGARRRGG SASRSLPLPK RPRRGAAPEP ERTPVGQGSW AHPGRTRGPS DRGFCVVSPA RPAEEATSL EGALSGTRHS HPSVGRQHHA GPPSTSRPPR PWDTPCPPVY AETKHFLYS SGDKEQLRPS FLLSSLRPSL TGARRLVETI FLGSRPWMPG TPRRLPRLPQ RYWQMRPL F LELLGNHAQC PYGVLLKTHC PLRAAVTPAA GVCAREKPQG SVAAPEEEDT DPRRLVQ LL RQHSSPWQVY GFVRACLRRL VPPGLWGSRH NERRFLRNTK KFISLGKHAK LSLQEL TWK MSVRDCAWLR RSPGVGCVPA AEHRLREEIL AKFLHWLMSV YVVELLRSFF YVTET TFQK NRLFFYRKSV WSKLQSIGIR QHLKRVQLRE LSEAEVRQHR EARPALLTSR LRFI PKPDG LRPIVNMDYV VGARTFRREK RAERLTSRVK ALFSVLNYER ARRPGLLGAS VLG LDDIHR AWRTFVLRVR AQDPPPELYF VKVDVTGAYD TIPQDRLTEV IASIIKPQNT YC VRRYAVV QKAAHGHVRK AFKSHVSTLT DLQPYMRQFV AHLQETSPLR DAVVIEQSSS L NEASSGLF DVFLRFMCHH AVRIRGKSYV QCQGIPQGSI LSTLLCSLCY GDMENKLFAG IRRDGLLLR LVDDFLLVTP HLTHAKTFLR TLVRGVPEYG CVVNLRKTVV NFPVEDEAL GGTAFVQMPA HGLFPWCGLL LDTRTLEVQS DYSSYARTSI RASLTFNRGF KAGRNMRR K LFGVLRLKCH SLFLDLQVNS LQTVCTNIYK ILLLQAYRFH ACVLQLPFHQ QVWKNPT FF LRVISDTASL CYSILKAKNA GMSLGAKGAA GPLPSEAVQW LCHQAFLLKL TRHRVT YVP LLGSLRTAQT QLSRKLPGTT LTALEAAANP ALPSDFKTIL D |
-Macromolecule #2: Histone H2A
Macromolecule | Name: Histone H2A / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) / Organ: Kidney / Tissue: Kidney |
Sequence | String: MSGRGKQGGK ARAKAKTRSS RAGLQFPVGR VRRLLRKGNY AERVGAGAPV YLAAVLEYLT AEILELAGN AARDNKKTRI IPRHLQLAIR NDEELNKLLG KVTIAQGGVL PNIQAVLLPK K TESHHKAK GK |
-Macromolecule #3: Histone H2B
Macromolecule | Name: Histone H2B / type: protein_or_peptide / ID: 3 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) / Organ: Kidney / Tissue: Kidney |
Sequence | String: MPDPAKSAPA PKKGSKKAVT KVQKKDGKKR KRSRKESYSV YVYKVLKQVH PDTGISSKAM GIMNSFVND IFERIAGEAS RLAHYNKRST ITSREIQTAV RLLLPGELAK HAVSEGTKAV T KYTSSNPR NLSPTKPGGS EDRQPPPSQL SAIPPFCLVL RAGIAGQV |
-Macromolecule #4: TPP1
Macromolecule | Name: TPP1 / type: protein_or_peptide / ID: 4 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Recombinant expression | Organism: Spodoptera (butterflies/moths) |
Sequence | String: MAGSGRLVLR PWIRELILGS ETPSSPRAGQ LLEVLQDAEA AVAGPSHAPD TSDVGATLLV SDGTHSVRC LVTREALDTS DWEEKEFGFR GTEGRLLLLQ DCGVHVQVAE GGAPAEFYLQ V DRFSLLPT EQPRLRVPGC NQDLDVQKKL YDCLEEHLSE STSSNAGLSL ...String: MAGSGRLVLR PWIRELILGS ETPSSPRAGQ LLEVLQDAEA AVAGPSHAPD TSDVGATLLV SDGTHSVRC LVTREALDTS DWEEKEFGFR GTEGRLLLLQ DCGVHVQVAE GGAPAEFYLQ V DRFSLLPT EQPRLRVPGC NQDLDVQKKL YDCLEEHLSE STSSNAGLSL SQLLDEMRED QE HQGALVC LAESCLTLEG PCTAPPVTHW AASRCKATGE AVYTVPSSML CISENDQLIL SSL GPCQRT QGPELPPPDP ALQDLSLTLI ASPPSSPSSS GTPALPGHMS SEESGTSISL LPAL SLAAP DPGQRSSSQP SPAICSAPAT LTPRSPHASR TPSSPLQSCT PSLSPRSHVP SPHQA LVTR PQKPSLEFKE FVGLPCKNRP PFPRTGATRG AQEPCSVWEP PKRHRDGSAF QYEYEP PCT SLCARVQAVR LPPQLMAWAL HFLMDAQPGS EPTPM |
-Macromolecule #5: human telomerase RNA
Macromolecule | Name: human telomerase RNA / type: rna / ID: 5 |
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Source (natural) | Organism: Homo sapiens (human) |
Sequence | String: GGGUUGCGGA GGGUGGGCCU GGGAGGGGUG GUGGCCAUUU UUUGUCUAAC CCUAACUGAG AAGGGCGUAG GCGCCGUGC UUUUGCUCCC CGCGCGCUGU UUUUCUCGCU GACUUUCAGC GGGCGGAAAA GCCUCGGCCU G CCGCCUUC CACCGUUCAU UCUAGAGCAA ...String: GGGUUGCGGA GGGUGGGCCU GGGAGGGGUG GUGGCCAUUU UUUGUCUAAC CCUAACUGAG AAGGGCGUAG GCGCCGUGC UUUUGCUCCC CGCGCGCUGU UUUUCUCGCU GACUUUCAGC GGGCGGAAAA GCCUCGGCCU G CCGCCUUC CACCGUUCAU UCUAGAGCAA ACAAAAAAUG UCAGCUGCUG GCCCGUUCGC CCCUCCCGGG GA CCUGCGG CGGGUCGCCU GCCCAGCCCC CGAACCCCGC CUGGAGGCCG CGGUCGGCCC GGGGCUUCUC CGG AGGCAC CCACUGCCAC CGCGAAGAGU UGGGCUCUGU CAGCCGCGGG UCUCUCGGGG GCGAGGGCGA GGUU CAGGC CUUUCAGGCC GCAGGAAGAG GAACGGAGCG AGUCCCCGCG CGCGGCGCGA UUCCCUGAGC UGUGG GACG UGCACCCAGG ACUCGGCUCA CACAUGC |
-Macromolecule #6: Telomeric DNA
Macromolecule | Name: Telomeric DNA / type: dna / ID: 6 / Classification: DNA |
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Source (natural) | Organism: Homo sapiens (human) |
Sequence | String: TTAGGGTTAG GGTTAGGGTT AGGGTTAGGG |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 8 Component:
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Grid | Model: C-flat / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 5 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 15 sec. / Pretreatment - Atmosphere: AIR | ||||||||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Quantum LS / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Digitization - Dimensions - Width: 5760 pixel / Digitization - Dimensions - Height: 4092 pixel / Number grids imaged: 3 / Number real images: 50775 / Average exposure time: 3.0 sec. / Average electron dose: 48.0 e/Å2 Details: Images were collected in movie-mode and fractionated into 48 movie frames |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 50.0 µm / Calibrated magnification: 45872 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 81000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Initial model | PDB ID: Chain - Source name: PDB / Chain - Initial model type: experimental model |
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Refinement | Space: RECIPROCAL / Protocol: AB INITIO MODEL |