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- EMDB-63833: Cryo-EM structure of spMETTL16 in complex with U6 snRNA_delta17 -
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Open data
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Basic information
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Title | Cryo-EM structure of spMETTL16 in complex with U6 snRNA_delta17 | |||||||||
![]() | Processed by DeepEMhancer. | |||||||||
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![]() | m6A methyltransferase / U6 snRNA / protein-RNA complex / splicing / TRANSFERASE / TRANSFERASE-RNA complex | |||||||||
Function / homology | ![]() snRNA (adenine-N6)-methylation / U6 snRNA m6A methyltransferase / U6 snRNA (adenine-(43)-N(6))-methyltransferase activity / rRNA base methylation / mRNA splicing, via spliceosome / nucleus / cytosol Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.4 Å | |||||||||
![]() | Ju J / Tomita K | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structures and mechanisms of U6 snRNA mA modification by METTL16. Authors: Jue Ju / Kozo Tomita / ![]() Abstract: The N-methyladenosine (mA) modification in U6 snRNA, catalyzed by METTL16 using S-adenosylmethionine (SAM) as the methyl donor, is required for efficient and accurate pre-mRNA splicing. However, the ...The N-methyladenosine (mA) modification in U6 snRNA, catalyzed by METTL16 using S-adenosylmethionine (SAM) as the methyl donor, is required for efficient and accurate pre-mRNA splicing. However, the mechanism by which METTL16 modifies U6 snRNA with mA remains elusive. Here, we present cryo-EM structures of METTL16 in complex with U6 snRNA, providing insights into the METTL16-mediated modification of U6 snRNA with mA. The structures reveal that U6 snRNA is recruited to METTL16 through specific interactions between the C-terminal kinase-associated 1 (KA-1) domain of METTL16 and the internal stem-loop (ISL) of U6 snRNA. Upon SAM binding to the catalytic pocket within the N-terminal methyltransferase domain (MTD), U6 snRNA undergoes a structural rearrangement that positions the target adenine-containing motif at the catalytic site. This conformational change is followed by an additional structural adjustment of U6 snRNA into a productive conformation, bringing the target adenosine closer to SAM within the catalytic pocket and thereby ensuring efficient mA modification. The KA-1 domain functions as a scaffold for initial substrate recognition and facilitates the subsequent dynamic methylation process within the MTD, highlighting the cooperative roles of METTL16 domains for U6 snRNA modification. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 81.1 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 21.1 KB 21.1 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 9.5 KB | Display | ![]() |
Images | ![]() | 43.8 KB | ||
Masks | ![]() | 91.1 MB | ![]() | |
Filedesc metadata | ![]() | 7 KB | ||
Others | ![]() ![]() | 84.5 MB 84.5 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 662.3 KB | Display | ![]() |
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Full document | ![]() | 661.9 KB | Display | |
Data in XML | ![]() | 17.6 KB | Display | |
Data in CIF | ![]() | 22.9 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 9u47MC ![]() 9m86C ![]() 9u48C 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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Annotation | Processed by DeepEMhancer. | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.83 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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Density Histograms |
-Half map: #2
File | emd_63833_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
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Density Histograms |
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Sample components
-Entire : Schizosaccharomyces pombe N6-methyladenosine methyltransferase ME...
Entire | Name: Schizosaccharomyces pombe N6-methyladenosine methyltransferase METTL16 in complex with U6 snRNA_delta17 |
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Components |
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-Supramolecule #1: Schizosaccharomyces pombe N6-methyladenosine methyltransferase ME...
Supramolecule | Name: Schizosaccharomyces pombe N6-methyladenosine methyltransferase METTL16 in complex with U6 snRNA_delta17 type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() ![]() |
-Macromolecule #1: U6 small nuclear RNA
Macromolecule | Name: U6 small nuclear RNA / type: rna / ID: 1 / Number of copies: 1 |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 26.476871 KDa |
Sequence | String: GGUCAAAUUG AAACGAUACA GAGAAGAUUA GCAUGGCCCC UGCACAAGGA UGACACUGCG ACAUUGAGAG AAAACCCAUU UU GENBANK: GENBANK: 3JB9_N |
-Macromolecule #2: U6 small nuclear RNA (adenine-(43)-N(6))-methyltransferase
Macromolecule | Name: U6 small nuclear RNA (adenine-(43)-N(6))-methyltransferase type: protein_or_peptide / ID: 2 Details: Residues MET A -22 to SER A 0 are expression tags. MET A 1 to PHE A 23 are part of the protein sequence, based on publication (DOI: 10.1038/s41467-021-23457-6). Number of copies: 1 / Enantiomer: LEVO / EC number: U6 snRNA m6A methyltransferase |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 48.442223 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MGSSHHHHHH SSGLVPRGSH MASMTDKTDI INFDSLARDY PDLRSFVKNG RIDFWNEDAI RTLGKAILDR DYSLRVEFPE NRLCPMVPN RATYIRYIHD LLSSTSGQKD KKRIIGLDIG TGASCIYPLL GCRMYSYDFV GTEIDKFSFE TAKSNILQNN M ESQIKIVL ...String: MGSSHHHHHH SSGLVPRGSH MASMTDKTDI INFDSLARDY PDLRSFVKNG RIDFWNEDAI RTLGKAILDR DYSLRVEFPE NRLCPMVPN RATYIRYIHD LLSSTSGQKD KKRIIGLDIG TGASCIYPLL GCRMYSYDFV GTEIDKFSFE TAKSNILQNN M ESQIKIVL RSKQDCLLPD TEGMEEFTFV MCNPPFYEHE EDFINFKQNP PSGVCTGVYH EMVTEGGEVG FANKILTESK KR KGIQWYT CMFGKKSSVP AVVDKLREQN ISNYGIYELA LGKTKRWIIC WSFQAMRPHN ELIRPSSTSL SKYFPHKVLQ NWT LDPELC AQIDDILQKF LDDNKIPWSK KGSVLEISTK SITWSRKARR ISKSQTSVSS LEGQMKCELN VIDNQLQCKW IEGY DYNVY ESFCSALARA LRDNKK UniProtKB: U6 small nuclear RNA (adenine-(43)-N(6))-methyltransferase |
-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 |
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Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Digitization - Dimensions - Width: 4092 pixel / Digitization - Dimensions - Height: 5760 pixel / Number grids imaged: 1 / Number real images: 6010 / Average exposure time: 1.0 sec. / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 1.8 µm / Nominal defocus min: 0.8 µm / Nominal magnification: 105000 |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Initial model | Chain - Source name: AlphaFold / Chain - Initial model type: in silico model |
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Output model | ![]() PDB-9u47: |