+Open data
-Basic information
Entry | Database: PDB / ID: 7uy7 | ||||||||||||
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Title | Tetrahymena CST with Polymerase alpha-Primase | ||||||||||||
Components |
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Keywords | REPLICATION / telomerase / RNP / CST / polymerase | ||||||||||||
Function / homology | Function and homology information alpha DNA polymerase:primase complex / lagging strand elongation / mitotic DNA replication initiation / telomerase holoenzyme complex / leading strand elongation / DNA replication origin binding / telomere maintenance via telomerase / single-stranded DNA binding / chromosome, telomeric region / DNA-directed DNA polymerase ...alpha DNA polymerase:primase complex / lagging strand elongation / mitotic DNA replication initiation / telomerase holoenzyme complex / leading strand elongation / DNA replication origin binding / telomere maintenance via telomerase / single-stranded DNA binding / chromosome, telomeric region / DNA-directed DNA polymerase / DNA-directed DNA polymerase activity / nucleotide binding / chromatin binding / metal ion binding Similarity search - Function | ||||||||||||
Biological species | Tetrahymena thermophila (eukaryote) | ||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 4.2 Å | ||||||||||||
Authors | He, Y. / Song, H. / Chan, H. / Wang, Y. / Liu, B. / Susac, L. / Zhou, Z.H. / Feigon, J. | ||||||||||||
Funding support | United States, 3items
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Citation | Journal: Nature / Year: 2022 Title: Structure of Tetrahymena telomerase-bound CST with polymerase α-primase. Authors: Yao He / He Song / Henry Chan / Baocheng Liu / Yaqiang Wang / Lukas Sušac / Z Hong Zhou / Juli Feigon / Abstract: Telomeres are the physical ends of linear chromosomes. They are composed of short repeating sequences (such as TTGGGG in the G-strand for Tetrahymena thermophila) of double-stranded DNA with a single- ...Telomeres are the physical ends of linear chromosomes. They are composed of short repeating sequences (such as TTGGGG in the G-strand for Tetrahymena thermophila) of double-stranded DNA with a single-strand 3' overhang of the G-strand and, in humans, the six shelterin proteins: TPP1, POT1, TRF1, TRF2, RAP1 and TIN2. TPP1 and POT1 associate with the 3' overhang, with POT1 binding the G-strand and TPP1 (in complex with TIN2) recruiting telomerase via interaction with telomerase reverse transcriptase (TERT). The telomere DNA ends are replicated and maintained by telomerase, for the G-strand, and subsequently DNA polymerase α-primase (PolαPrim), for the C-strand. PolαPrim activity is stimulated by the heterotrimeric complex CTC1-STN1-TEN1 (CST), but the structural basis of the recruitment of PolαPrim and CST to telomere ends remains unknown. Here we report cryo-electron microscopy (cryo-EM) structures of Tetrahymena CST in the context of the telomerase holoenzyme, in both the absence and the presence of PolαPrim, and of PolαPrim alone. Tetrahymena Ctc1 binds telomerase subunit p50, a TPP1 orthologue, on a flexible Ctc1 binding motif revealed by cryo-EM and NMR spectroscopy. The PolαPrim polymerase subunit POLA1 binds Ctc1 and Stn1, and its interface with Ctc1 forms an entry port for G-strand DNA to the POLA1 active site. We thus provide a snapshot of four key components that are required for telomeric DNA synthesis in a single active complex-telomerase-core ribonucleoprotein, p50, CST and PolαPrim-that provides insights into the recruitment of CST and PolαPrim and the handoff between G-strand and C-strand synthesis. | ||||||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 7uy7.cif.gz | 369.2 KB | Display | PDBx/mmCIF format |
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PDB format | pdb7uy7.ent.gz | 280 KB | Display | PDB format |
PDBx/mmJSON format | 7uy7.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7uy7_validation.pdf.gz | 1.2 MB | Display | wwPDB validaton report |
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Full document | 7uy7_full_validation.pdf.gz | 1.2 MB | Display | |
Data in XML | 7uy7_validation.xml.gz | 67.9 KB | Display | |
Data in CIF | 7uy7_validation.cif.gz | 100.4 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/uy/7uy7 ftp://data.pdbj.org/pub/pdb/validation_reports/uy/7uy7 | HTTPS FTP |
-Related structure data
Related structure data | 26866MC 7uy5C 7uy6C 7uy8C M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
-Telomerase-associated protein of ... , 3 types, 3 molecules ABC
#1: Protein | Mass: 73899.602 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Tetrahymena thermophila (eukaryote) / References: UniProt: A0PGB2 |
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#2: Protein | Mass: 43682.938 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Tetrahymena thermophila (eukaryote) / References: UniProt: Q6JXI5 |
#3: Protein | Mass: 19498.049 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Tetrahymena thermophila (eukaryote) / References: UniProt: D2CVN7 |
-Protein , 2 types, 2 molecules DE
#4: Protein | Mass: 50049.688 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Tetrahymena thermophila (eukaryote) / References: UniProt: D2CVN8 |
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#5: Protein | Mass: 161986.984 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Tetrahymena thermophila (eukaryote) / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: Q23AJ0, DNA-directed DNA polymerase |
-DNA chain / Non-polymers , 2 types, 2 molecules F
#6: DNA chain | Mass: 19207.121 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) Tetrahymena thermophila (eukaryote) |
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#7: Chemical | ChemComp-ZN / |
-Details
Has ligand of interest | Y |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component |
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Source (natural) |
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Source (recombinant) | Organism: Spodoptera frugiperda (fall armyworm) | ||||||||||||||||||||||||
Buffer solution | pH: 8 | ||||||||||||||||||||||||
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | ||||||||||||||||||||||||
Vitrification | Cryogen name: ETHANE |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 4000 nm / Nominal defocus min: 800 nm |
Image recording | Electron dose: 55 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
-Processing
Software | Name: PHENIX / Version: 1.19.2_4158: / Classification: refinement | ||||||||||||||||||||||||
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
3D reconstruction | Resolution: 4.2 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 142912 / Symmetry type: POINT | ||||||||||||||||||||||||
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