[English] 日本語
![](img/lk-miru.gif)
- EMDB-29001: Cryo-EM structure of Cascade-PAM complex in type I-B CAST system -
+
Open data
-
Basic information
Entry | ![]() | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Cryo-EM structure of Cascade-PAM complex in type I-B CAST system | |||||||||
![]() | ||||||||||
![]() |
| |||||||||
![]() | ![]() ![]() ![]() | |||||||||
Function / homology | ![]() | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | ![]() ![]() | |||||||||
![]() | Chang L / Wang S | |||||||||
Funding support | ![]()
| |||||||||
![]() | ![]() 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 |
|
-
Structure visualization
Supplemental images |
---|
-
Downloads & links
-EMDB archive
Map data | ![]() | 58.9 MB | ![]() | |
---|---|---|---|---|
Header (meta data) | ![]() ![]() | 20.9 KB 20.9 KB | Display Display | ![]() |
Images | ![]() | 115.5 KB | ||
Filedesc metadata | ![]() | 6.5 KB | ||
Others | ![]() ![]() | 59.4 MB 59.4 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8fd3MC ![]() 8fcjC ![]() 8fcuC ![]() 8fcvC ![]() 8fcwC ![]() 8fcxC ![]() 8fd2C ![]() 8ff4C ![]() 8ff5C M: atomic model generated by this map C: citing same article ( |
---|---|
Similar structure data | Similarity search - Function & homology ![]() |
-
Links
EMDB pages | ![]() ![]() |
---|---|
Related items in Molecule of the Month |
-
Map
File | ![]() | ||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Voxel size | X=Y=Z: 1.054 Å | ||||||||||||||||||||
Density |
| ||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
|
-Supplemental data
-Half map: #2
File | emd_29001_half_map_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: #1
File | emd_29001_half_map_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-
Sample components
-Entire : Ternary complex of Cascade proteins, crRNA and PAM only DNA
Entire | Name: Ternary complex of Cascade proteins, crRNA and PAM only DNA |
---|---|
Components |
|
-Supramolecule #1: Ternary complex of Cascade proteins, crRNA and PAM only DNA
Supramolecule | Name: Ternary complex of Cascade proteins, crRNA and PAM only DNA type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
---|---|
Source (natural) | Organism: ![]() |
-Macromolecule #1: Type I-B CRISPR-associated protein Cas5
Macromolecule | Name: Type I-B CRISPR-associated protein Cas5 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 24.852906 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MIAPLILYLD VPFTTFRESH AREMGKTYPV PPPATVYGML LSLVGETNVY RHCGVELAIA MLSSPKKSRI LRQMRRFKNA DFSHPENVI PCYQEILSNL KCLIWVRSDE EKIQPSLRER IQLAFDHPEL VRRFGCLFLG ESDQLIKTIK LAREDYLEGV R QWAIRDNR ...String: MIAPLILYLD VPFTTFRESH AREMGKTYPV PPPATVYGML LSLVGETNVY RHCGVELAIA MLSSPKKSRI LRQMRRFKNA DFSHPENVI PCYQEILSNL KCLIWVRSDE EKIQPSLRER IQLAFDHPEL VRRFGCLFLG ESDQLIKTIK LAREDYLEGV R QWAIRDNR GRLTLPYWVD HVGSRNTRFL RYRIEEMDRL SPPDLAWTMV QSPI UniProtKB: Type I-MYXAN CRISPR-associated protein Cas5/Cmx5/DevS |
-Macromolecule #2: Type I-B CRISPR-associated protein Cas6
Macromolecule | Name: Type I-B CRISPR-associated protein Cas6 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 24.945744 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MTAILIQSEE YVDLTFKLRG APIPLDNGYL TYAALSRICP PLHELKSIGI HPIAGIPTRN NLLELTAQSR LKIRIYHQQI PLIYPYLAG QAFHIGQNFY QLDIPDYKPL ISSESVYSRL VIIKGFQDST NFIEAVQRQM DNLGIQGKIE LLTRQDGTPQ R RQLTINKE ...String: MTAILIQSEE YVDLTFKLRG APIPLDNGYL TYAALSRICP PLHELKSIGI HPIAGIPTRN NLLELTAQSR LKIRIYHQQI PLIYPYLAG QAFHIGQNFY QLDIPDYKPL ISSESVYSRL VIIKGFQDST NFIEAVQRQM DNLGIQGKIE LLTRQDGTPQ R RQLTINKE GKQFKVRGFG VKISELNPED SLTLQEQGIG GKRKMMCGIF VPATRSKEEE ET UniProtKB: Type I-MYXAN CRISPR-associated protein Cas6/Cmx6 |
-Macromolecule #3: Type I-B CRISPR-associated protein Cas7
Macromolecule | Name: Type I-B CRISPR-associated protein Cas7 / type: protein_or_peptide / ID: 3 / Number of copies: 6 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 37.298996 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MMTQKKNDSN IPNYYLYGTV LTRYGLASLN HDIRRGNKTI LQKGYWNNGK IHSFVGSSAI RWALRFYLQK QGYLVNRVWD EEEHINRLT SEDFDPEKFY DDDIFGFALL ESAETEEDTS TTKRKKKQTK TSTPNQRMGA LGMNMAVSLT PYDGAVKLGA K SGREKDST ...String: MMTQKKNDSN IPNYYLYGTV LTRYGLASLN HDIRRGNKTI LQKGYWNNGK IHSFVGSSAI RWALRFYLQK QGYLVNRVWD EEEHINRLT SEDFDPEKFY DDDIFGFALL ESAETEEDTS TTKRKKKQTK TSTPNQRMGA LGMNMAVSLT PYDGAVKLGA K SGREKDST SLHFTEYHAT RYQYYFGIDA THLKDFSRIL PMIDGIMNLP KVGGSSNIFN YPFCPDSLVF QWTNHFASYI SY CFEYCDP KSKEAKLSQE FIDEVECGQI DPSKLWIGGT IVKDLQQLDN FESSPLNKAH IYRNRNEMIE ALKTVIKRDL GLE ESK UniProtKB: CRISPR-associated protein |
-Macromolecule #4: Type I-MYXAN CRISPR-associated Cas8a1/Cmx1
Macromolecule | Name: Type I-MYXAN CRISPR-associated Cas8a1/Cmx1 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 63.474328 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MHHHHHHHHI VSTQPKISLS LHAADTTIMH RVGMTGLYMT LKRLEKQYPL SRQRGGHISW FLTADTIELF WEGSDFIALS WLINESFQL DDTGLIHLVG LDNDRIDLRQ KIHIHEGICG VFLRLNKFYQ AGEIINTELR FEEKQVEYQY KSLTWYAHQT F AEKLCEAD ...String: MHHHHHHHHI VSTQPKISLS LHAADTTIMH RVGMTGLYMT LKRLEKQYPL SRQRGGHISW FLTADTIELF WEGSDFIALS WLINESFQL DDTGLIHLVG LDNDRIDLRQ KIHIHEGICG VFLRLNKFYQ AGEIINTELR FEEKQVEYQY KSLTWYAHQT F AEKLCEAD TQQLRHDYIQ ITSWLYLGGI VRHARTQNTT KLEEKPEYAL ALLFVPVVCH YCLLHIPSED LKERKPHRYL VV IPEIKDF EDASQRRWRL QQLETKQFHV SSLGEAGLLY YSLDDIQPEV AYYQACQVWL YEKTNKASRQ RTLMSIEEIK IDK NILITY QQVQKYFKTN YQIIKYKQIF IKVNPIRSLI ADNLVKGIHW WSNFWEKLVI EDSKEYLFNQ LFSNREGFII MAEN SEEDK QYLIFIKVFQ QAMKGNFAKI YAKTEEGKDP PIKKKVERLR AELNYCYDEL SFKEYLSDFL VRGGLNKYFN EHQEE IALL IKKSPWQEIR IWSLLAIASY KPKDKLTNRD DSSLSNNQKL EEVNDDSEEE UniProtKB: Type I-MYXAN CRISPR-associated Cas8a1/Cmx1 |
-Macromolecule #5: Type I-B CRISPR-associated protein Cas11
Macromolecule | Name: Type I-B CRISPR-associated protein Cas11 / type: protein_or_peptide / ID: 5 / Number of copies: 3 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 16.314365 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MAENSEEDKQ YLIFIKVFQQ AMKGNFAKIY AKTEEGKDPP IKKKVERLRA ELNYCYDELS FKEYLSDFLV RGGLNKYFNE HQEEIALLI KKSPWQEIRI WSLLAIASYK PKDKLTNRDD SSLSNNQKLE EVNDDSEEE UniProtKB: Type I-MYXAN CRISPR-associated Cas8a1/Cmx1 |
-Macromolecule #6: RNA
Macromolecule | Name: RNA / type: rna / ID: 6 / Number of copies: 1 |
---|---|
Source (natural) | Organism: synthetic construct (others) |
Molecular weight | Theoretical: 22.876527 KDa |
Sequence | String: UUGCUCAAGA GAAGUCAUUU AAUAAGGCCA CUGUUAAACG UAGGUGAGUC GUGGCUUUAU GCCGUUAGGC G |
-Macromolecule #7: Target DNA strand
Macromolecule | Name: Target DNA strand / type: dna / ID: 7 / Number of copies: 1 / Classification: DNA |
---|---|
Source (natural) | Organism: synthetic construct (others) |
Molecular weight | Theoretical: 3.003993 KDa |
Sequence | String: (DC)(DA)(DT)(DG)(DA)(DT)(DC)(DT)(DA)(DC) |
-Macromolecule #8: Non-target DNA strand
Macromolecule | Name: Non-target DNA strand / type: dna / ID: 8 / Number of copies: 1 / Classification: DNA |
---|---|
Source (natural) | Organism: synthetic construct (others) |
Molecular weight | Theoretical: 13.973018 KDa |
Sequence | String: (DG)(DT)(DA)(DG)(DA)(DT)(DC)(DA)(DT)(DG) (DG)(DA)(DG)(DA)(DA)(DG)(DT)(DC)(DA)(DT) (DT)(DT)(DA)(DA)(DT)(DA)(DA)(DG)(DG) (DC)(DC)(DA)(DC)(DT)(DG)(DT)(DT)(DA)(DA) (DA) (DC)(DG)(DT)(DA)(DG) |
-Experimental details
-Structure determination
Method | ![]() |
---|---|
![]() | ![]() |
Aggregation state | particle |
-
Sample preparation
Buffer | pH: 7.5 |
---|---|
Vitrification | Cryogen name: ETHANE |
-
Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
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 K3 BIOQUANTUM (6k x 4k) / Average electron dose: 54.0 e/Å2 |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
-
Image processing
Startup model | Type of model: PDB ENTRY PDB model - PDB ID: |
---|---|
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |
Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.12 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 204496 |