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- EMDB-28994: Cryo-EM structure of TnsC-TniQ-DNA complex in type I-B CAST system -
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Open data
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Basic information
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Title | Cryo-EM structure of TnsC-TniQ-DNA complex in type I-B CAST system | |||||||||
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![]() | TnsC / TniQ / CRISPR / DNA BINDING PROTEIN | |||||||||
Function / homology | TniQ / TniQ / : / AAA domain / ATP hydrolysis activity / P-loop containing nucleoside triphosphate hydrolase / ORC1/DEAH AAA+ ATPase domain-containing protein / TniQ family protein![]() | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.95 Å | |||||||||
![]() | Chang L / Wang S | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Molecular mechanism for Tn7-like transposon recruitment by a type I-B CRISPR effector. Authors: Shukun Wang / Clinton Gabel / Romana Siddique / Thomas Klose / Leifu Chang / ![]() Abstract: Tn7-like transposons have co-opted CRISPR-Cas systems to facilitate the movement of their own DNA. These CRISPR-associated transposons (CASTs) are promising tools for programmable gene knockin. A key ...Tn7-like transposons have co-opted CRISPR-Cas systems to facilitate the movement of their own DNA. These CRISPR-associated transposons (CASTs) are promising tools for programmable gene knockin. A key feature of CASTs is their ability to recruit Tn7-like transposons to nuclease-deficient CRISPR effectors. However, how Tn7-like transposons are recruited by diverse CRISPR effectors remains poorly understood. Here, we present the cryo-EM structure of a recruitment complex comprising the Cascade complex, TniQ, TnsC, and the target DNA in the type I-B CAST from Peltigera membranacea cyanobiont 210A. Target DNA recognition by Cascade induces conformational changes in Cas6 and primes TniQ recruitment through its C-terminal domain. The N-terminal domain of TniQ is bound to the seam region of the TnsC spiral heptamer. Our findings provide insights into the diverse mechanisms for the recruitment of Tn7-like transposons to CRISPR effectors and will aid in the development of CASTs as gene knockin tools. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 30.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 17 KB 17 KB | Display Display | ![]() |
Images | ![]() | 122.2 KB | ||
Filedesc metadata | ![]() | 5.9 KB | ||
Others | ![]() ![]() | 59.3 MB 59.3 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8fcvMC ![]() 8fcjC ![]() 8fcuC ![]() 8fcwC ![]() 8fcxC ![]() 8fd2C ![]() 8fd3C ![]() 8ff4C ![]() 8ff5C M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Map
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Voxel size | X=Y=Z: 1.054 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: #2
File | emd_28994_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_28994_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Ternary complex of TnsC, TniQ and DNA in type I-B CAST system
Entire | Name: Ternary complex of TnsC, TniQ and DNA in type I-B CAST system |
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Components |
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-Supramolecule #1: Ternary complex of TnsC, TniQ and DNA in type I-B CAST system
Supramolecule | Name: Ternary complex of TnsC, TniQ and DNA in type I-B CAST system type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#4 |
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Source (natural) | Organism: ![]() |
-Macromolecule #1: DNA (60-MER)
Macromolecule | Name: DNA (60-MER) / type: dna / ID: 1 / Number of copies: 1 / Classification: DNA |
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Source (natural) | Organism: synthetic construct (others) |
Molecular weight | Theoretical: 18.500861 KDa |
Sequence | String: (DA)(DA)(DG)(DA)(DC)(DG)(DT)(DA)(DG)(DC) (DC)(DC)(DA)(DG)(DC)(DG)(DC)(DG)(DT)(DC) (DG)(DG)(DC)(DC)(DG)(DC)(DT)(DA)(DC) (DG)(DT)(DA)(DT)(DC)(DG)(DT)(DA)(DG)(DA) (DT) (DA)(DT)(DA)(DT)(DC)(DT) ...String: (DA)(DA)(DG)(DA)(DC)(DG)(DT)(DA)(DG)(DC) (DC)(DC)(DA)(DG)(DC)(DG)(DC)(DG)(DT)(DC) (DG)(DG)(DC)(DC)(DG)(DC)(DT)(DA)(DC) (DG)(DT)(DA)(DT)(DC)(DG)(DT)(DA)(DG)(DA) (DT) (DA)(DT)(DA)(DT)(DC)(DT)(DA)(DC) (DG)(DC)(DG)(DT)(DA)(DG)(DA)(DT)(DA)(DT) (DA)(DT) |
-Macromolecule #2: DNA (60-MER)
Macromolecule | Name: DNA (60-MER) / type: dna / ID: 2 / Number of copies: 1 / Classification: DNA |
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Source (natural) | Organism: synthetic construct (others) |
Molecular weight | Theoretical: 18.482832 KDa |
Sequence | String: (DA)(DT)(DA)(DT)(DA)(DT)(DC)(DT)(DA)(DC) (DG)(DC)(DG)(DT)(DA)(DG)(DA)(DT)(DA)(DT) (DA)(DT)(DC)(DT)(DA)(DC)(DG)(DA)(DT) (DA)(DC)(DG)(DT)(DA)(DG)(DC)(DG)(DG)(DC) (DC) (DG)(DA)(DC)(DG)(DC)(DG) ...String: (DA)(DT)(DA)(DT)(DA)(DT)(DC)(DT)(DA)(DC) (DG)(DC)(DG)(DT)(DA)(DG)(DA)(DT)(DA)(DT) (DA)(DT)(DC)(DT)(DA)(DC)(DG)(DA)(DT) (DA)(DC)(DG)(DT)(DA)(DG)(DC)(DG)(DG)(DC) (DC) (DG)(DA)(DC)(DG)(DC)(DG)(DC)(DT) (DG)(DG)(DG)(DC)(DT)(DA)(DC)(DG)(DT)(DC) (DT)(DT) |
-Macromolecule #3: TniQ
Macromolecule | Name: TniQ / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 37.591961 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MTIYSEHWSL NPLEIPQRSR LFSLEPVAVG TPYAESLSSY LHRLAQAHCL TSEKLVMGEI APLILKDEDK SELLSKNLSH LLGNSDAKP AINGMREMTE KLVTVLEELT MRQDLRFLTL LSWKGMIYDK GLFRNYRAWC PCCCEEWMQK NKTIYEPLSW S FKDVEFCL ...String: MTIYSEHWSL NPLEIPQRSR LFSLEPVAVG TPYAESLSSY LHRLAQAHCL TSEKLVMGEI APLILKDEDK SELLSKNLSH LLGNSDAKP AINGMREMTE KLVTVLEELT MRQDLRFLTL LSWKGMIYDK GLFRNYRAWC PCCCEEWMQK NKTIYEPLSW S FKDVEFCL IHKQRLIEEC SHCGARLPVM ARLSPAGFCS RCYGWLGQEI KGEEEIEKYR VNIQGISELI ALTPQLGYKP IP IELTRKL QLILLVFEQA IGKDVKLLGD LGGIMESLRI ASTTNQSQPY HLVKLIIPVC EKAKISVFQL FGSDFKELGK ILF GNFSLE LKL UniProtKB: TniQ family protein |
-Macromolecule #4: TnsC
Macromolecule | Name: TnsC / type: protein_or_peptide / ID: 4 / Number of copies: 7 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 43.521934 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MTKSTGFPLE LLTRPATERL AYFENYTVAH PRLKEVYEIL MRTIAEPAGA SFIFVYGASG VGKTTLRLRV EQKLTELALP KLESDRARV PVVGIEAIAP ESRYFNWKEY YTRALITLEE PLIDHKFDYG VRGISRDNFG KINVESKVVA PALRRALENA L IHRHPDVF ...String: MTKSTGFPLE LLTRPATERL AYFENYTVAH PRLKEVYEIL MRTIAEPAGA SFIFVYGASG VGKTTLRLRV EQKLTELALP KLESDRARV PVVGIEAIAP ESRYFNWKEY YTRALITLEE PLIDHKFDYG VRGISRDNFG KINVESKVVA PALRRALENA L IHRHPDVF FVDEAQHFGK VASGYKLQDQ LDCLKSLANM TGILHCLLGT YELLTFRNLS GQLSRRSVDI HFRRYCADSP ED VQAFKSV LLTFQQHLPL AETPNLVDHW EYFYERTLGC IGTLKDWLKR VLSDALDREA TTITLKDLQK RALSVAQCQK MFK EIQEGE RQLSETEADV QNLRSALGLG AKPIVLPEET PKTTRPPGKV GKRKPKRDPI GVQQDVS UniProtKB: ORC1/DEAH AAA+ ATPase domain-containing protein |
-Macromolecule #5: ADENOSINE-5'-TRIPHOSPHATE
Macromolecule | Name: ADENOSINE-5'-TRIPHOSPHATE / type: ligand / ID: 5 / Number of copies: 7 / Formula: ATP |
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Molecular weight | Theoretical: 507.181 Da |
Chemical component information | ![]() ChemComp-ATP: |
-Macromolecule #6: MAGNESIUM ION
Macromolecule | Name: MAGNESIUM ION / type: ligand / ID: 6 / Number of copies: 7 / Formula: MG |
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Molecular weight | Theoretical: 24.305 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 BIOQUANTUM (6k x 4k) / Average electron dose: 54.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.8 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Startup model | Type of model: INSILICO MODEL |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 2.95 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 185087 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |