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Yorodumi- EMDB-50241: cryoEM structure of Asgard tubulin heterodimer AtubA/B2 with GTP -
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Open data
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Basic information
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| Title | cryoEM structure of Asgard tubulin heterodimer AtubA/B2 with GTP | |||||||||
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 Keywords | cryoEM / tubulin / Asgard archaea / microtubule / CYTOSOLIC PROTEIN | |||||||||
| Biological species |  Candidatus Lokiarchaeum ossiferum (archaea) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.5 Å | |||||||||
 Authors | Wollweber F / Xu J / Ponce-Toled RI / Rodrigues-Oliveira T / Malit JJL / Kokhanovska A / Schleper C / Pilhofer M | |||||||||
| Funding support | European Union, 1 items 
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 Citation |  Journal: Cell / Year: 2025Title: Microtubules in Asgard archaea. Authors: Florian Wollweber / Jingwei Xu / Rafael I Ponce-Toledo / Florina Marxer / Thiago Rodrigues-Oliveira / Anja Pössnecker / Zhen-Hao Luo / Jessie James Limlingan Malit / Anastasiia Kokhanovska ...Authors: Florian Wollweber / Jingwei Xu / Rafael I Ponce-Toledo / Florina Marxer / Thiago Rodrigues-Oliveira / Anja Pössnecker / Zhen-Hao Luo / Jessie James Limlingan Malit / Anastasiia Kokhanovska / Michal Wieczorek / Christa Schleper / Martin Pilhofer /   ![]() Abstract: Microtubules are a hallmark of eukaryotes. Archaeal and bacterial homologs of tubulins typically form homopolymers and non-tubular superstructures. The origin of heterodimeric tubulins assembling ...Microtubules are a hallmark of eukaryotes. Archaeal and bacterial homologs of tubulins typically form homopolymers and non-tubular superstructures. The origin of heterodimeric tubulins assembling into microtubules remains unclear. Here, we report the discovery of microtubule-forming tubulins in Asgard archaea, the closest known relatives of eukaryotes. These Asgard tubulins (AtubA/B) are closely related to eukaryotic α/β-tubulins and the enigmatic bacterial tubulins BtubA/B. Proteomics of Candidatus Lokiarchaeum ossiferum showed that AtubA/B were highly expressed. Cryoelectron microscopy structures demonstrate that AtubA/B form eukaryote-like heterodimers, which assembled into 5-protofilament bona fide microtubules in vitro. The additional paralog AtubB2 lacks a nucleotide-binding site and competitively displaced AtubB. These AtubA/B2 heterodimers polymerized into 7-protofilament non-canonical microtubules. In a sub-population of Ca. Lokiarchaeum ossiferum cells, cryo-tomography revealed tubular structures, while expansion microscopy identified AtubA/B cytoskeletal assemblies. Our findings suggest a pre-eukaryotic origin of microtubules and provide a framework for understanding the fundamental principles of microtubule assembly.  | |||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data |  emd_50241.map.gz | 21 MB |  EMDB map data format | |
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| Header (meta data) |  emd-50241-v30.xml emd-50241.xml | 19.4 KB 19.4 KB  | Display Display  |  EMDB header | 
| Images |  emd_50241.png | 25.6 KB | ||
| Masks |  emd_50241_msk_1.map | 22.2 MB |  Mask map | |
| Filedesc metadata |  emd-50241.cif.gz | 6.3 KB | ||
| Others |  emd_50241_additional_1.map.gz emd_50241_half_map_1.map.gz emd_50241_half_map_2.map.gz | 20.1 MB 20.6 MB 20.6 MB  | ||
| Archive directory |  http://ftp.pdbj.org/pub/emdb/structures/EMD-50241 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-50241 | HTTPS FTP  | 
-Validation report
| Summary document |  emd_50241_validation.pdf.gz | 806.2 KB | Display |  EMDB validaton report | 
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| Full document |  emd_50241_full_validation.pdf.gz | 805.8 KB | Display | |
| Data in XML |  emd_50241_validation.xml.gz | 10.2 KB | Display | |
| Data in CIF |  emd_50241_validation.cif.gz | 12 KB | Display | |
| Arichive directory |  https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-50241 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-50241 | HTTPS FTP  | 
-Related structure data
| Related structure data | ![]() 9f6vMC ![]() 9f6tC ![]() 9f6uC ![]() 9hxcC M: atomic model generated by this map C: citing same article (  | 
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Links
| EMDB pages |  EMDB (EBI/PDBe) /  EMDataResource | 
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Map
| File |  Download / File: emd_50241.map.gz / Format: CCP4 / Size: 22.2 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Projections & slices | Image control
 
 Images are generated by Spider.  | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.3 Å | ||||||||||||||||||||||||||||||||||||
| Density | 
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML: 
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-Supplemental data
-Mask #1
| File |  emd_50241_msk_1.map | ||||||||||||
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| Density Histograms | 
-Additional map: the map that was processed by DeepEMhancer.
| File | emd_50241_additional_1.map | ||||||||||||
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| Annotation | the map that was processed by DeepEMhancer. | ||||||||||||
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| Density Histograms | 
-Half map: #1
| File | emd_50241_half_map_1.map | ||||||||||||
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| Projections & Slices | 
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| Density Histograms | 
-Half map: #2
| File | emd_50241_half_map_2.map | ||||||||||||
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| Density Histograms | 
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Sample components
-Entire : Asgard tubulin heterodimer AtubA/B2 from Candidatus Lokiarchaeum ...
| Entire | Name: Asgard tubulin heterodimer AtubA/B2 from Candidatus Lokiarchaeum ossiferum | 
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| Components | 
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-Supramolecule #1: Asgard tubulin heterodimer AtubA/B2 from Candidatus Lokiarchaeum ...
| Supramolecule | Name: Asgard tubulin heterodimer AtubA/B2 from Candidatus Lokiarchaeum ossiferum type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2  | 
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| Source (natural) | Organism:  Candidatus Lokiarchaeum ossiferum (archaea) | 
-Macromolecule #1: Asgard tubulin A (AtubA) from Candidatus Lokiarchaeum ossiferum
| Macromolecule | Name: Asgard tubulin A (AtubA) from Candidatus Lokiarchaeum ossiferum type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO  | 
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| Source (natural) | Organism:  Candidatus Lokiarchaeum ossiferum (archaea) | 
| Molecular weight | Theoretical: 46.040281 KDa | 
| Recombinant expression | Organism: ![]()  | 
| Sequence | String: MAGEIVCIQV GQAGNQIAGA FWQKICAEHG IDPVNGKAID VVGDTDIFFN TIGDKYIPRA VVVDLEPAVV ENIREKFGTL  FDPKSIVSG ADGAGNNFAI GFNEHGAETL EKVMQVVEQR VSETESIGGF ILTHSCGGGT GSGFGSKILK TIRERYPKVP I FTFSIFPS  ...String:  MAGEIVCIQV GQAGNQIAGA FWQKICAEHG IDPVNGKAID VVGDTDIFFN TIGDKYIPRA VVVDLEPAVV ENIREKFGTL  FDPKSIVSG ADGAGNNFAI GFNEHGAETL EKVMQVVEQR VSETESIGGF ILTHSCGGGT GSGFGSKILK TIRERYPKVP I FTFSIFPS PKISETVVEP YNAIMTLSNL IKYASCSIVL DNEALFSIAE KKLEVENPSL EDLNLIIAQV LTNVTASLRF SG TLNLDLG KLVTNLVPFS NLHFLMASTA PLVLAGKESY EKMTAKELSA QVFGDEYICA ACKPTTGRYL AASVLFRGAV KTS DVNEAM ATVKEQNSFV NWIPTGFKIS KSETSPKDSA LGVIMLGNNS EIVSVFERIG ANFDRLWSRK AFAHWFTDSG FEEK DLDDA RALVQKVIDD YRKLTEDA  | 
-Macromolecule #2: Asgard tubulin B homolog (AtubB2) from Candidatus Lokiarchaeum os...
| Macromolecule | Name: Asgard tubulin B homolog (AtubB2) from Candidatus Lokiarchaeum ossiferum type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO  | 
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| Source (natural) | Organism:  Candidatus Lokiarchaeum ossiferum (archaea) | 
| Molecular weight | Theoretical: 47.882949 KDa | 
| Recombinant expression | Organism: ![]()  | 
| Sequence | String: MAREVITIHV GELGIQIAPN FWKYLCDEHN IDYKGQEKGK IRGVIDNFFE KASIGKWIPR TILVDLGPNA IRKVTKKDMK  DFFDPKRCV MGLAGDANLF AKGYYSYGTR FMEEIMDKIQ KEVDQTEHLQ GFIVVHSIGD GTGAGLAPLI MEAIKKKHPK L VMMSYSIV  ...String:  MAREVITIHV GELGIQIAPN FWKYLCDEHN IDYKGQEKGK IRGVIDNFFE KASIGKWIPR TILVDLGPNA IRKVTKKDMK  DFFDPKRCV MGLAGDANLF AKGYYSYGTR FMEEIMDKIQ KEVDQTEHLQ GFIVVHSIGD GTGAGLAPLI MEAIKKKHPK L VMMSYSIV PSQNMDCSTI LPYNAILSLD KLTSCADISM IIDNDSIYRI VATQGKENEL SESIFDQVLA KALVEITATL RF NSPLNRS MMEMSTNLVP FPRNHFLMTS MSPLETSLTS AHQKIETKEL MQDLIDQDHI LAPITVEKGV FTAFVIALRG ENP HSILQN SIKGFGDRVK FSEIFPTAIK ADSTTLTDEK LARSGITLMN HSGVANLFQF LLTQFELMYD HDAFTTWYYQ EGMQ PSEFE AAKNNIQKLI TEYKQDEY  | 
-Macromolecule #3: GUANOSINE-5'-TRIPHOSPHATE
| Macromolecule | Name: GUANOSINE-5'-TRIPHOSPHATE / type: ligand / ID: 3 / Number of copies: 1 / Formula: GTP | 
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| Molecular weight | Theoretical: 523.18 Da | 
| Chemical component information | ![]() ChemComp-GTP:   | 
-Experimental details
-Structure determination
| Method | cryo EM | 
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 Processing | single particle reconstruction | 
| Aggregation state | particle | 
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Sample preparation
| Buffer | pH: 6.8 | 
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| Grid | Model: UltrAuFoil / Material: GOLD / Support film - Material: GRAPHENE OXIDE / Support film - topology: CONTINUOUS | 
| Vitrification | Cryogen name: ETHANE-PROPANE | 
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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: 60.0 e/Å2 | 
| Electron beam | Acceleration voltage: 300 kV / Electron source:  FIELD EMISSION GUN | 
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.8000000000000003 µm / Nominal defocus min: 1.2 µm | 
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company  | 
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About Yorodumi



Keywords
Candidatus Lokiarchaeum ossiferum (archaea)
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Processing
FIELD EMISSION GUN
