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Open data
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Basic information
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| Title | In-situ cryo-EM structure of porinI of the Dot/Icm machine | |||||||||
Map data | structure of porinI of the Dot/Icm machine | |||||||||
Sample |
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Keywords | Type IVB Dot/Icm Secretion Machine / PROTEIN TRANSPORT | |||||||||
| Function / homology | Function and homology information | |||||||||
| Biological species | Legionella pneumophila subsp. pneumophila (bacteria) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.56 Å | |||||||||
Authors | Yue J / Liu J | |||||||||
| Funding support | United States, 1 items
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Citation | Journal: bioRxiv / Year: 2025 Title: structures of the Dot/Icm T4SS identify the DotA-IcmX complex as the gatekeeper for effector translocation. Authors: Jian Yue / Samira Heydari / Donghyun Park / David Chetrit / Shoichi Tachiyama / Wangbiao Guo / Jack M Botting / Shenping Wu / Craig R Roy / Jun Liu Abstract: The Dot/Icm machine in is one of the most versatile type IV secretion systems (T4SSs), with a remarkable capacity to translocate over 330 different effector proteins across the bacterial envelope ...The Dot/Icm machine in is one of the most versatile type IV secretion systems (T4SSs), with a remarkable capacity to translocate over 330 different effector proteins across the bacterial envelope into host cells. At least 27 Dot and Icm proteins are required for assembly and function of the system, yet the architecture and activation mechanism remain poorly understood at the molecular level. Here, we deploy cryo-electron microscopy to reveal structures of the Dot/Icm machine at near-atomic resolution. Importantly, two proteins essential for effector translocation, DotA and IcmX, form a pentameric protochannel at an inactive state. Upon activation, the DotA-IcmX protochannel undergoes extensive rearrangements to form an extended transenvelope passage capable of transporting effector proteins from the bacterial cytoplasm into host cells as revealed by cryo-electron tomography. Collectively, our findings suggest that the DotA-IcmX complex functions as the gatekeeper for effector translocation of the Dot/Icm T4SS. | |||||||||
| History |
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Structure visualization
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_49399.map.gz | 28.5 MB | EMDB map data format | |
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| Header (meta data) | emd-49399-v30.xml emd-49399.xml | 18.4 KB 18.4 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_49399_fsc.xml | 6.6 KB | Display | FSC data file |
| Images | emd_49399.png | 85 KB | ||
| Filedesc metadata | emd-49399.cif.gz | 5.9 KB | ||
| Others | emd_49399_half_map_1.map.gz emd_49399_half_map_2.map.gz | 27.5 MB 27.5 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-49399 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-49399 | HTTPS FTP |
-Validation report
| Summary document | emd_49399_validation.pdf.gz | 916.2 KB | Display | EMDB validaton report |
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| Full document | emd_49399_full_validation.pdf.gz | 916 KB | Display | |
| Data in XML | emd_49399_validation.xml.gz | 14 KB | Display | |
| Data in CIF | emd_49399_validation.cif.gz | 18.1 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-49399 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-49399 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9nh1MC ![]() 9nguC ![]() 9ngvC ![]() 9ngwC ![]() 9ngyC ![]() 9nh0C ![]() 9nh2C M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_49399.map.gz / Format: CCP4 / Size: 30.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | structure of porinI of the Dot/Icm machine | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.068 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Half map: Half Map A
| File | emd_49399_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_49399_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 : porinI
| Entire | Name: porinI |
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| Components |
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-Supramolecule #1: porinI
| Supramolecule | Name: porinI / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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| Source (natural) | Organism: Legionella pneumophila subsp. pneumophila (bacteria) |
-Macromolecule #1: Major outer membrane protein
| Macromolecule | Name: Major outer membrane protein / type: protein_or_peptide / ID: 1 / Number of copies: 3 / Enantiomer: LEVO |
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| Source (natural) | Organism: Legionella pneumophila subsp. pneumophila (bacteria) |
| Molecular weight | Theoretical: 32.155463 KDa |
| Sequence | String: MFSLKKTAVA VLALGSGAVF AGTMGPVCTP GNVTVPCERT AWDIGITALY LQPIYDADWG YNGFTQVGGW RHWHDVDHEW DWGFKLEGS YHFNTGNDIN VNWYHFDNDS DHWFDFANWH NYNNKWDAVN AELGQFVDFS ANKKMRFHGG VQYARIEADV N RYFNNFAF ...String: MFSLKKTAVA VLALGSGAVF AGTMGPVCTP GNVTVPCERT AWDIGITALY LQPIYDADWG YNGFTQVGGW RHWHDVDHEW DWGFKLEGS YHFNTGNDIN VNWYHFDNDS DHWFDFANWH NYNNKWDAVN AELGQFVDFS ANKKMRFHGG VQYARIEADV N RYFNNFAF NGFNSKFNGF GPRTGLDMNY VFGNGFGIYA KGAAAILVGT SDFYDGIGFV TGSKNAIVPE LEAKLGADYT YA MAQGDLT LDVGYMWFNY FNAMHNTATN GLETDFAASG PYIGLKYVGN V UniProtKB: Major outer membrane protein |
-Macromolecule #2: Outer membrane protein, OmpA family protein
| Macromolecule | Name: Outer membrane protein, OmpA family protein / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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| Source (natural) | Organism: Legionella pneumophila subsp. pneumophila (bacteria) |
| Molecular weight | Theoretical: 27.582047 KDa |
| Sequence | String: MRNLMRCLIM IKSLIKGVDM SRKLAKTRIL GYGLMICFLA GCFHPPYNNF QPDRRAVKRV GVDTGIGAVA GAIASGTASG TLIGAAAGG TVGLVASIYR DSKRKIIRDL QKQDIQYVEY GDTRTLIIPT DKYFMFSSPR LNEICYPGLN NVIRLLNFYP Q STIYVAGF ...String: MRNLMRCLIM IKSLIKGVDM SRKLAKTRIL GYGLMICFLA GCFHPPYNNF QPDRRAVKRV GVDTGIGAVA GAIASGTASG TLIGAAAGG TVGLVASIYR DSKRKIIRDL QKQDIQYVEY GDTRTLIIPT DKYFMFSSPR LNEICYPGLN NVIRLLNFYP Q STIYVAGF TDNVGSRSHK RKLSQAQAET MMTFLWANGI AAKRLKAEGY GDKNAISDNA IIHGSAQNRR IEIQWFTSPA QP PQPQMAY VK UniProtKB: Outer membrane protein, OmpA family protein |
-Macromolecule #3: LphA (DotK)
| Macromolecule | Name: LphA (DotK) / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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| Source (natural) | Organism: Legionella pneumophila subsp. pneumophila (bacteria) |
| Molecular weight | Theoretical: 21.096492 KDa |
| Sequence | String: MRSLRTNYIY VLFKTTGLLF LLLLSACNRS GYIPENEVPK LPCRVDGACD ATIIKMMTDL NKKGIKVASV GQNYLISIPA SALFADQSP RLNWASYSLL NEIAAFLKQF RKIAITVTSY SSKYVSVKRE RALTLARSRV VSEYLWSQGV DSRIIFTQGL G SDKPITSY TLGGDRSPNA RVEITFRRAV A UniProtKB: LphA (DotK) |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | cell |
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Sample preparation
| Buffer | pH: 6.7 |
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| Vitrification | Cryogen name: ETHANE |
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Electron microscopy
| Microscope | FEI TECNAI 12 |
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| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 73.0 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: 2.0 µm / Nominal defocus min: 0.8 µm |
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Keywords
Legionella pneumophila subsp. pneumophila (bacteria)
Authors
United States, 1 items
Citation
















Z (Sec.)
Y (Row.)
X (Col.)




































Processing
FIELD EMISSION GUN
