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- EMDB-4303: Cryo-EM subtomogram average of IFT complex B and inhibited dynein... -
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Basic information
Entry | Database: EMDB / ID: EMD-4303 | |||||||||
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Title | Cryo-EM subtomogram average of IFT complex B and inhibited dynein-1b in anterograde IFT trains (Chlamydomonas reinhardtii) | |||||||||
![]() | Cryo-EM stubtomogram average of IFT complex B and inhibited dynein-1b in anterograde IFT trains (Chlamydomonas reinhardtii) | |||||||||
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Function / homology | ![]() intraciliary transport involved in cilium assembly / nitric-oxide synthase inhibitor activity / deoxyribonuclease inhibitor activity / negative regulation of DNA strand resection involved in replication fork processing / organelle organization / visual behavior / intraciliary retrograde transport / methylated-DNA-[protein]-cysteine S-methyltransferase / methylated-DNA-[protein]-cysteine S-methyltransferase activity / intraciliary transport ...intraciliary transport involved in cilium assembly / nitric-oxide synthase inhibitor activity / deoxyribonuclease inhibitor activity / negative regulation of DNA strand resection involved in replication fork processing / organelle organization / visual behavior / intraciliary retrograde transport / methylated-DNA-[protein]-cysteine S-methyltransferase / methylated-DNA-[protein]-cysteine S-methyltransferase activity / intraciliary transport / dynein light chain binding / ciliary transition zone / regulation of cilium assembly / motile cilium assembly / dynein heavy chain binding / Activation of BIM and translocation to mitochondria / embryonic skeletal system morphogenesis / ciliary tip / Intraflagellar transport / negative regulation of nitric oxide biosynthetic process / dynein complex / cell projection organization / negative regulation of phosphorylation / COPI-independent Golgi-to-ER retrograde traffic / minus-end-directed microtubule motor activity / dynein light intermediate chain binding / cytoplasmic dynein complex / ciliary plasm / motile cilium / enzyme inhibitor activity / determination of left/right symmetry / microtubule motor activity / ciliary base / dynein intermediate chain binding / Macroautophagy / pericentriolar material / microtubule-based movement / axoneme / positive regulation of insulin secretion involved in cellular response to glucose stimulus / tertiary granule membrane / ficolin-1-rich granule membrane / spermatid development / cilium assembly / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / COPI-mediated anterograde transport / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / Loss of Nlp from mitotic centrosomes / Loss of proteins required for interphase microtubule organization from the centrosome / Recruitment of mitotic centrosome proteins and complexes / axon cytoplasm / Recruitment of NuMA to mitotic centrosomes / Anchoring of the basal body to the plasma membrane / Resolution of Sister Chromatid Cohesion / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / substantia nigra development / MHC class II antigen presentation / centriole / AURKA Activation by TPX2 / ciliary basal body / filopodium / RHO GTPases Activate Formins / cilium / mitotic spindle / kinetochore / Aggrephagy / HCMV Early Events / Separation of Sister Chromatids / Regulation of PLK1 Activity at G2/M Transition / apical part of cell / site of double-strand break / scaffold protein binding / methylation / microtubule / cytoskeleton / DNA repair / centrosome / DNA damage response / Neutrophil degranulation / protein-containing complex binding / apoptotic process / Golgi apparatus / enzyme binding / ATP hydrolysis activity / mitochondrion / DNA binding / extracellular space / ATP binding / identical protein binding / membrane / nucleus / metal ion binding / plasma membrane / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 37.0 Å | |||||||||
![]() | Pigino G / Jordan MA | |||||||||
![]() | ![]() Title: The cryo-EM structure of intraflagellar transport trains reveals how dynein is inactivated to ensure unidirectional anterograde movement in cilia. Authors: Mareike A Jordan / Dennis R Diener / Ludek Stepanek / Gaia Pigino / ![]() Abstract: Movement of cargos along microtubules plays key roles in diverse cellular processes, from signalling to mitosis. In cilia, rapid movement of ciliary components along the microtubules to and from the ...Movement of cargos along microtubules plays key roles in diverse cellular processes, from signalling to mitosis. In cilia, rapid movement of ciliary components along the microtubules to and from the assembly site is essential for the assembly and disassembly of the structure itself. This bidirectional transport, known as intraflagellar transport (IFT), is driven by the anterograde motor kinesin-2 and the retrograde motor dynein-1b (dynein-2 in mammals). However, to drive retrograde transport, dynein-1b must first be delivered to the ciliary tip by anterograde IFT. Although, the presence of opposing motors in bidirectional transport processes often leads to periodic stalling and slowing of cargos, IFT is highly processive. Using cryo-electron tomography, we show that a tug-of-war between kinesin-2 and dynein-1b is prevented by loading dynein-1b onto anterograde IFT trains in an autoinhibited form and by positioning it away from the microtubule track to prevent binding. Once at the ciliary tip, dynein-1b must transition into an active form and engage microtubules to power retrograde trains. These findings provide a striking example of how coordinated structural changes mediate the behaviour of complex cellular machinery. | |||||||||
History |
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Structure visualization
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 168.2 KB | ![]() | |
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Header (meta data) | ![]() ![]() | 8.9 KB 8.9 KB | Display Display | ![]() |
Images | ![]() ![]() | 284.4 KB 188 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6sc2M ![]() 4304C M: atomic model generated by this map C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Cryo-EM stubtomogram average of IFT complex B and inhibited dynein-1b in anterograde IFT trains (Chlamydomonas reinhardtii) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 14.13 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Intraflagellar transport complex B and inhibited dynein-1b
Entire | Name: Intraflagellar transport complex B and inhibited dynein-1b |
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Components |
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-Supramolecule #1: Intraflagellar transport complex B and inhibited dynein-1b
Supramolecule | Name: Intraflagellar transport complex B and inhibited dynein-1b type: complex / ID: 1 / Parent: 0 |
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Source (natural) | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | subtomogram averaging |
Aggregation state | 2D array |
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Sample preparation
Buffer | pH: 7 |
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Grid | Model: Quantifoil R3.5/1 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE / Instrument: LEICA EM GP |
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Electron microscopy
Microscope | FEI TITAN |
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Specialist optics | Energy filter - Name: GIF Quantum LS |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Average electron dose: 2.1 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
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Image processing
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 37.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: PEET (ver. 1.11.0) / Number subtomograms used: 750 |
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Extraction | Number tomograms: 15 / Number images used: 750 / Software - Name: ![]() |
CTF correction | Software - Name: CTFPHASEFLIP |
Final angle assignment | Type: NOT APPLICABLE |