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Open data
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Basic information
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Title | Tetrahymena Polymerase alpha-Primase | ||||||||||||
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![]() | CST / polymerase / primase / REPLICATION | ||||||||||||
Function / homology | ![]() alpha DNA polymerase:primase complex / DNA primase activity / lagging strand elongation / DNA replication, synthesis of primer / mitotic DNA replication initiation / leading strand elongation / DNA replication origin binding / Transferases; Transferring phosphorus-containing groups; Nucleotidyltransferases / single-stranded DNA binding / 4 iron, 4 sulfur cluster binding ...alpha DNA polymerase:primase complex / DNA primase activity / lagging strand elongation / DNA replication, synthesis of primer / mitotic DNA replication initiation / leading strand elongation / DNA replication origin binding / Transferases; Transferring phosphorus-containing groups; Nucleotidyltransferases / single-stranded DNA binding / 4 iron, 4 sulfur cluster binding / DNA replication / DNA-directed DNA polymerase / DNA-directed DNA polymerase activity / nucleotide binding / chromatin binding / DNA binding / nucleus / metal ion binding Similarity search - Function | ||||||||||||
Biological species | ![]() ![]() | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.5 Å | ||||||||||||
![]() | He Y / Song H / Chan H / Wang Y / Liu B / Susac L / Zhou ZH / Feigon J | ||||||||||||
Funding support | ![]()
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![]() | ![]() 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
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 49.6 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 23.9 KB 23.9 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 9.1 KB | Display | ![]() |
Images | ![]() | 50.7 KB | ||
Masks | ![]() | 64 MB | ![]() | |
Filedesc metadata | ![]() | 7.4 KB | ||
Others | ![]() ![]() ![]() | 49.1 MB 49.7 MB 49.7 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 667.3 KB | Display | ![]() |
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Full document | ![]() | 666.9 KB | Display | |
Data in XML | ![]() | 16.1 KB | Display | |
Data in CIF | ![]() | 21.1 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7uy8MC ![]() 7uy5C ![]() 7uy6C ![]() 7uy7C C: citing same article ( M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homology ![]() |
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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 | full map | ||||||||||||||||||||
Voxel size | X=Y=Z: 1.1 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
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-Additional map: composite map generated using two focused refined maps, sharpened
File | emd_26867_additional_1.map | ||||||||||||
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Annotation | composite map generated using two focused refined maps, sharpened | ||||||||||||
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-Half map: half1
File | emd_26867_half_map_1.map | ||||||||||||
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Annotation | half1 | ||||||||||||
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-Half map: half2
File | emd_26867_half_map_2.map | ||||||||||||
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Annotation | half2 | ||||||||||||
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Sample components
-Entire : Tetrahymena PolaPrim
Entire | Name: Tetrahymena PolaPrim |
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Components |
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-Supramolecule #1: Tetrahymena PolaPrim
Supramolecule | Name: Tetrahymena PolaPrim / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#4 |
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Source (natural) | Organism: ![]() ![]() |
-Macromolecule #1: DNA polymerase alpha subunit B
Macromolecule | Name: DNA polymerase alpha subunit B / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 68.773758 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MEEFEKILEE IDSANLSQNA VQQLKQLIIS NGILDDEQLM EKWLFQLEAI IYNSKQKFLE AQLMDFKKNV LNQDIKKPQN DNFKKIKES EVQKNIDIIK GDADLFTKAS SAKKQTSTQQ ANNAAIDCEE QLRQQLGKES MLDPKRFSYN SNLPQVNFEK N QHQNSNNM ...String: MEEFEKILEE IDSANLSQNA VQQLKQLIIS NGILDDEQLM EKWLFQLEAI IYNSKQKFLE AQLMDFKKNV LNQDIKKPQN DNFKKIKES EVQKNIDIIK GDADLFTKAS SAKKQTSTQQ ANNAAIDCEE QLRQQLGKES MLDPKRFSYN SNLPQVNFEK N QHQNSNNM TIVDLFEQYD PNKIPNYLTN NIDKIRKHLE QRILTFKIQN YSQFIVNGEQ LINDVSSYSK TEEVKMVGRL IS TENGFLN TTNLRIELNR NQYFDLVFEN DFDFGNNVLF PNQIVMVKGI INEKEELVVS ELITDHIKEI TDEEHPKIDN LNQ DESQLI MVAAGPFSTV MSTQYTSFDN ILHVAKTKKV STLILLGPFL DIKNEILKDG SIIINSKEYT FEQLQNSLFE KAVK ELEGK TQIIIVPSTR DILSTDSIPQ MSLEIKVSEH KNSIHSYSNP AYIDIDKLRV YIANSDVGMI TLQNSLLDKK IPYMQ QSRL TFKALMNQQN LYSIYPMRNP QDSQQILETV KLPNINDFDI PFDYQLEQYP HIIISPSSLP KFATKIYNTV VINPNY VIR DGSQAGNFAI ITLFKDSDIP IHERTRVDLY CL UniProtKB: DNA polymerase alpha subunit B |
-Macromolecule #2: DNA polymerase
Macromolecule | Name: DNA polymerase / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed DNA polymerase |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 161.986984 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MSDKLTRLER LNKEVKKQNK LKQHSKNNRF DDDMDIEAYE DDEQIEEDDF IDDTQEDKKY KKKYREIEDE FDQEIEEEEE LNKKKKTKN TILNYTNTTA VTNNKKKAIS KQIPDIDIEE IMKLTERKKK IEQEEAQLLQ EEQELLEQEK REEEEKKRIS Q EAKSILRE ...String: MSDKLTRLER LNKEVKKQNK LKQHSKNNRF DDDMDIEAYE DDEQIEEDDF IDDTQEDKKY KKKYREIEDE FDQEIEEEEE LNKKKKTKN TILNYTNTTA VTNNKKKAIS KQIPDIDIEE IMKLTERKKK IEQEEAQLLQ EEQELLEQEK REEEEKKRIS Q EAKSILRE DCASSKTANS SKGKVDQNIL NAINRDFSDD SNTVDSISEF QKLKSLAQKA NLANESLKQS KVSNTEINLT NL SISQVKK INDYKNEDGS VDAYLYDYFY DAQVKPDKIY AFAKVQNKQT NAFDTCVIQI DTIIRNLFFY PSSDTVTEQQ IKN EIAELL KKEQTSRKNV EFLGAFVDKN YAFELPIPRG KSRWYQVVMS YEYEVISPDT KGQYFSYCVG STYSALETFL ITKK ITGPS WVRFQNVKDT TSCITNRKLE FRVDYTNQSN IQVLQKQLPT PPLSVVCISL KTSQQIVLSQ KKKEYKKEIF NLNMK YHEG INIDNSNKDE LNQFKSISFI THIDPTKKQD SITKKGTLPE TTKFCLNELN LLEQFLVHFN EIDPDIVVAH DLYSTV FEI ILTRIREKGI RKWNLLSKLI NIGSSDIPKY GSSTFKTKMA MKGRLLVDTL LSSQEFVNCV EYTLEALAQK LFKIEIP RI DAKAYQQKFA TYKLLNSLVD DTYQDIDYAL RIMYHLQIVP LTKQLTSICG NIWMGSLQNQ RAERNEMLLL HKFNQLNY V YPDNFKNLPE SYKKKHKNAQ IRKQYEEDED QAQGNKNPKK KENKYKGGQV FEPEKGLYNE YIVLLDFNSL YPSIIQEFN VCFTTCVRDP IPLEMQMAPF LGNKKAAIQY SKNQNTKENK MQDEDEEDNE NEQIVQTHDV LPTIEVIKGI APLPSILQYL VEQRKVVKN QIKGQKDPQV IETLDIKQKA FKLVANSMYG CLGFSSSRFY AMPLASFITA KGRHILFDSK KIVEDMGYSV I YGDTDSLM IKPGTNEFLE AVKTGLSIKI KVNSKYKKLQ LDIDGVFKNM LLLKKKKYAT LKVANWEEVK NTNAPEKLEK EI KGIDVVR RDWCQLSRDA GNKILEIILE SKSSENMLDD IKKYLIQLND DINQKNIKNS NYYITKRLTK RVDQYGEKNL PHV AVAQRS IQEKGIDPQT YVNQIISYII CKNEQSSRLV DKAYSPQEFI TQSKSLEIDL QYYKRFQLFE PIKRMLEVIE GINL QEIAS ILEVHYSVQH VSQNNELNAE NVLNLKSKRN QFLTSIPRVL VDCKKCDQTF LFLGILEENA DAASILKCKC GNDIY IQLK NKIALVVKEL IRNFEENAIQ IDNEEFEYTH QISLVGKAKQ QKMSSFTLNQ KLLSIQAMFD ITKEEQENTQ KVTIEK IKT IKKTLDDLLS KSQYNNLNLS NIFTSFGLLK UniProtKB: DNA polymerase |
-Macromolecule #3: Eukaryotic-type DNA primase, large subunit
Macromolecule | Name: Eukaryotic-type DNA primase, large subunit / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 64.386645 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MINNKQIQKI QQISKFQLKE YIEMQMQQHS LNKQIKEEER NYILLENAID NSNIQGFQTI QEIVQLAMQR MKMIKEIIQE EPRFNLIDP LSKPNYNRIR VANKFITVAI PKSDPEYELK NKQQYAKEVL SHFLCKMAYA FYAEPETWLE FARAEAFILM D KLKSGQHH ...String: MINNKQIQKI QQISKFQLKE YIEMQMQQHS LNKQIKEEER NYILLENAID NSNIQGFQTI QEIVQLAMQR MKMIKEIIQE EPRFNLIDP LSKPNYNRIR VANKFITVAI PKSDPEYELK NKQQYAKEVL SHFLCKMAYA FYAEPETWLE FARAEAFILM D KLKSGQHH ANFFSDENLK IKTISDELFK QAFPKIEATF SSIKIKKNDS EVEQINIKKD NFKMFKFIDH PSMLSNNDVV LH KGYIIIY KESTSKIVQN IFIERLLDEM RLLQLKFQNN GSKLDDDRLS FLKDLHKAEI FNDSTQFNST QIHHYELDRL AKR DMPACM TYLMYGLNQK LHLKHFGRLQ LGLFLKGAGL SLNEALTFWQ KKFSKTSADD FKKKYDYNIR HNYGKLGKQL DYTP MSCQK IIGFQPLKDE FHGCPYKTME SQQLKDFLKL SYNITDEQFV QVNIFKNQKQ YQLACKEVFK VLNDTRDKTK EQFYP YFDK VGNHPNAYFE QSLRMHEPQR FQKQDEEKKQ NKQNRNQNFS ANKQSTNKNN QMDLEF UniProtKB: Eukaryotic-type DNA primase, large subunit |
-Macromolecule #4: DNA primase
Macromolecule | Name: DNA primase / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO EC number: Transferases; Transferring phosphorus-containing groups; Nucleotidyltransferases |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 47.172855 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MEIETVEVAP QQEEEFKLDY DLLEEYYRSY FPVSQMVQWL SYPQDGDDTY FTRREFSFTL QNEVYLRYQM YNSEREFKNA LLQKVPEKI DIGAVYDRPG KKGDDIKAKE KEFVIDIDMT DYDHIRTCCS KAKICEKCWK FMRVACDLIS KSLDEDFGFQ H VLWVYSGR ...String: MEIETVEVAP QQEEEFKLDY DLLEEYYRSY FPVSQMVQWL SYPQDGDDTY FTRREFSFTL QNEVYLRYQM YNSEREFKNA LLQKVPEKI DIGAVYDRPG KKGDDIKAKE KEFVIDIDMT DYDHIRTCCS KAKICEKCWK FMRVACDLIS KSLDEDFGFQ H VLWVYSGR RGIHAWVCDK EIRKANDYTR ASIIDYLNIL VDNSIGSSYV KPSLLKMEKS HLIERNAMKQ LNQKNDDLKA EK ESEKVFV EIVLREQNLF MKKPEIILEF LAKRSENLSK EVEKEWKTLK TSEQRYEALK ELVSSEDKKK THYLLEELRI WLL YPRLDV NVSKSTNHLL KSPFCIHPKT GNVCVPFTTE EISTFDPFSV PNISTLTTEE GSSKMKNSLK IFNKFLENLK KDV UniProtKB: DNA primase |
-Macromolecule #5: ZINC ION
Macromolecule | Name: ZINC ION / type: ligand / ID: 5 / Number of copies: 1 / Formula: ZN |
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Molecular weight | Theoretical: 65.409 Da |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
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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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 55.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 4.0 µm / Nominal defocus min: 0.8 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |