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- EMDB-15690: Cryo-EM structure of the Tb ADAT2/3 deaminase in complex with tRNA -
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Open data
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Basic information
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Title | Cryo-EM structure of the Tb ADAT2/3 deaminase in complex with tRNA | |||||||||
![]() | Final non b factor sharpened cryosparc map | |||||||||
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![]() | ADAT / inosine / tRNA modification / deaminase / cryo-EM structure / Trypanosoma brucei / RNA BINDING PROTEIN | |||||||||
Function / homology | ![]() tRNA wobble adenosine to inosine editing / tRNA-specific adenosine-34 deaminase activity / Hydrolases; Acting on carbon-nitrogen bonds, other than peptide bonds; In cyclic amidines / hydrolase activity / zinc ion binding / nucleoplasm / cytoplasm Similarity search - Function | |||||||||
Biological species | ![]() ![]() ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||
![]() | Dolce LG / Tengo L / Weis F / Kowalinski E | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural basis for sequence-independent substrate selection by eukaryotic wobble base tRNA deaminase ADAT2/3. Authors: Luciano G Dolce / Aubree A Zimmer / Laura Tengo / Félix Weis / Mary Anne T Rubio / Juan D Alfonzo / Eva Kowalinski / ![]() ![]() ![]() Abstract: The essential deamination of adenosine A to inosine at the wobble base is the individual tRNA modification with the greatest effects on mRNA decoding, empowering a single tRNA to translate three ...The essential deamination of adenosine A to inosine at the wobble base is the individual tRNA modification with the greatest effects on mRNA decoding, empowering a single tRNA to translate three different codons. To date, many aspects of how eukaryotic deaminases specifically select their multiple substrates remain unclear. Here, using cryo-EM, we present the structure of a eukaryotic ADAT2/3 deaminase bound to a full-length tRNA, revealing that the enzyme distorts the anticodon loop, but in contrast to the bacterial enzymes, selects its substrate via sequence-independent contacts of eukaryote-acquired flexible or intrinsically unfolded motifs distal from the conserved catalytic core. A gating mechanism for substrate entry to the active site is identified. Our multi-step tRNA recognition model yields insights into how RNA editing by A deamination evolved, shaped the genetic code, and directly impacts the eukaryotic proteome. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 50.9 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 18.7 KB 18.7 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 13.8 KB | Display | ![]() |
Images | ![]() | 51.3 KB | ||
Filedesc metadata | ![]() | 6.2 KB | ||
Others | ![]() ![]() ![]() | 90.3 MB 95.7 MB 95.7 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8aw3MC 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
File | ![]() | ||||||||||||||||||||
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Annotation | Final non b factor sharpened cryosparc map | ||||||||||||||||||||
Voxel size | X=Y=Z: 0.81 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: deepEMhancer post processed map
File | emd_15690_additional_1.map | ||||||||||||
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Annotation | deepEMhancer post processed map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: half map A
File | emd_15690_half_map_1.map | ||||||||||||
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Annotation | half map A | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: half map B
File | emd_15690_half_map_2.map | ||||||||||||
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Annotation | half map B | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : ADAT2/3 in complex with tRNA
Entire | Name: ADAT2/3 in complex with tRNA |
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Components |
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-Supramolecule #1: ADAT2/3 in complex with tRNA
Supramolecule | Name: ADAT2/3 in complex with tRNA / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#3 |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 90 KDa |
-Macromolecule #1: RNA (75-MER)
Macromolecule | Name: RNA (75-MER) / type: rna / ID: 1 / Number of copies: 1 |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 24.132311 KDa |
Sequence | String: GGCCGCUUAG CACAGUGGCA GUGCACCACU CUCGUAAAGU GGGGGUCGCG AGUUCGAUUC UCGCAGUGGC CUCCA GENBANK: GENBANK: AC091702.3 |
-Macromolecule #2: Deaminase, putative
Macromolecule | Name: Deaminase, putative / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO EC number: Hydrolases; Acting on carbon-nitrogen bonds, other than peptide bonds; In cyclic amidines |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 23.547535 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MVQDTGKDTN LKGTAEANES VVYCDVFMQA ALKEATCALE EGEVPVGCVL VKADSSTAAQ AQAGDDLALQ KLIVARGRNA TNRKGHGLA HAEFVAVEEL LRQATAGTSE NIGGGGNSGA VSQDLADYVL YVVVEPCIMC AAMLLYNRVR KVYFGCTNPR F GGNGTVLS ...String: MVQDTGKDTN LKGTAEANES VVYCDVFMQA ALKEATCALE EGEVPVGCVL VKADSSTAAQ AQAGDDLALQ KLIVARGRNA TNRKGHGLA HAEFVAVEEL LRQATAGTSE NIGGGGNSGA VSQDLADYVL YVVVEPCIMC AAMLLYNRVR KVYFGCTNPR F GGNGTVLS VHNSYKGCSG EDAALIGYES CGGYRAEEAV VLLQQFYRRE NTNAPLGKRK RKD UniProtKB: Deaminase, putative |
-Macromolecule #3: Deaminase, putative
Macromolecule | Name: Deaminase, putative / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 40.600254 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: GPDSMEEVVV PEEPPKLVSA LATYVQQERL CTMFLSIANK LLPLKPHACH LKRIRRSSAT RVATAPMDGF AGGVICDKRD SSVATSTIS DGCERNSAAL GTPAAEKSHV LELLLSVGGP VDSSALKELE SAADTTVAVH RVWVPDRAPR SSAEEWTKWC Q IWPFATPK ...String: GPDSMEEVVV PEEPPKLVSA LATYVQQERL CTMFLSIANK LLPLKPHACH LKRIRRSSAT RVATAPMDGF AGGVICDKRD SSVATSTIS DGCERNSAAL GTPAAEKSHV LELLLSVGGP VDSSALKELE SAADTTVAVH RVWVPDRAPR SSAEEWTKWC Q IWPFATPK PRVPTQLSEC EVGSIQRIFR TVVMPLAKRL RTDETLGIAA VLVDPSDGYR VLVSSGEEHA LKRGNSAACL GY VSNSGCR KSNRVVLDHP VTFVLKEVTR KQCKDREVEG DASYLANGMD MFVSHEPCVM CSMALVHSRV RRVFYCFPNP VHG GLGSTV SIHAIQELNH HFRVFRCDSR WLSDPEGVSS DHDNPYWEDL TVP UniProtKB: Deaminase, putative |
-Macromolecule #4: ZINC ION
Macromolecule | Name: ZINC ION / type: ligand / ID: 4 / Number of copies: 2 / 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
Concentration | 0.07 mg/mL |
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Buffer | pH: 7.5 |
Grid | Model: UltrAuFoil R1.2/1.3 / Material: GOLD |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 55.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.8 µm / Nominal defocus min: 1.0 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |