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- EMDB-4154: Mechanism of microtubule minus-end recognition and protection by ... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-4154 | |||||||||||||||||||||
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Title | Mechanism of microtubule minus-end recognition and protection by CAMSAP proteins | |||||||||||||||||||||
![]() | CAMSAP3 CKK decorated 13pf microtubule asymmetric unit | |||||||||||||||||||||
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![]() | CAMSAP CKK Microtubule Tubulin / Structural protein | |||||||||||||||||||||
Function / homology | ![]() regulation of organelle organization / zonula adherens maintenance / microtubule minus-end / protein transport along microtubule / microtubule anchoring / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly / Intraflagellar transport / Carboxyterminal post-translational modifications of tubulin / Sealing of the nuclear envelope (NE) by ESCRT-III ...regulation of organelle organization / zonula adherens maintenance / microtubule minus-end / protein transport along microtubule / microtubule anchoring / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly / Intraflagellar transport / Carboxyterminal post-translational modifications of tubulin / Sealing of the nuclear envelope (NE) by ESCRT-III / Kinesins / Resolution of Sister Chromatid Cohesion / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / COPI-dependent Golgi-to-ER retrograde traffic / RHO GTPases activate IQGAPs / COPI-independent Golgi-to-ER retrograde traffic / COPI-mediated anterograde transport / microtubule minus-end binding / RHO GTPases Activate Formins / regulation of Golgi organization / establishment or maintenance of microtubule cytoskeleton polarity / MHC class II antigen presentation / cilium movement / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / Aggrephagy / epithelial cell-cell adhesion / The role of GTSE1 in G2/M progression after G2 checkpoint / zonula adherens / Separation of Sister Chromatids / Loss of Nlp from mitotic centrosomes / Recruitment of mitotic centrosome proteins and complexes / Loss of proteins required for interphase microtubule organization from the centrosome / Anchoring of the basal body to the plasma membrane / AURKA Activation by TPX2 / Recruitment of NuMA to mitotic centrosomes / Regulation of PLK1 Activity at G2/M Transition / Hedgehog 'off' state / negative regulation of microtubule depolymerization / establishment of epithelial cell apical/basal polarity / motile cilium / positive regulation of axon guidance / embryo development ending in birth or egg hatching / regulation of focal adhesion assembly / spectrin binding / regulation of microtubule polymerization / axoneme / microtubule-based process / regulation of microtubule cytoskeleton organization / cytoplasmic microtubule organization / regulation of cell migration / ciliary basal body / Hydrolases; Acting on acid anhydrides; Acting on GTP to facilitate cellular and subcellular movement / structural constituent of cytoskeleton / microtubule cytoskeleton organization / microtubule cytoskeleton / neuron projection development / actin filament binding / mitotic cell cycle / nervous system development / in utero embryonic development / microtubule / calmodulin binding / hydrolase activity / protein heterodimerization activity / GTPase activity / centrosome / GTP binding / metal ion binding / cytosol / cytoplasm Similarity search - Function | |||||||||||||||||||||
Biological species | ![]() ![]() ![]() ![]() | |||||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 5.3 Å | |||||||||||||||||||||
![]() | Akhmanova A / Moores CA | |||||||||||||||||||||
Funding support | ![]() ![]() ![]() ![]()
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![]() | ![]() Title: A structural model for microtubule minus-end recognition and protection by CAMSAP proteins. Authors: Joseph Atherton / Kai Jiang / Marcel M Stangier / Yanzhang Luo / Shasha Hua / Klaartje Houben / Jolien J E van Hooff / Agnel-Praveen Joseph / Guido Scarabelli / Barry J Grant / Anthony J ...Authors: Joseph Atherton / Kai Jiang / Marcel M Stangier / Yanzhang Luo / Shasha Hua / Klaartje Houben / Jolien J E van Hooff / Agnel-Praveen Joseph / Guido Scarabelli / Barry J Grant / Anthony J Roberts / Maya Topf / Michel O Steinmetz / Marc Baldus / Carolyn A Moores / Anna Akhmanova / ![]() ![]() ![]() ![]() Abstract: CAMSAP and Patronin family members regulate microtubule minus-end stability and localization and thus organize noncentrosomal microtubule networks, which are essential for cell division, polarization ...CAMSAP and Patronin family members regulate microtubule minus-end stability and localization and thus organize noncentrosomal microtubule networks, which are essential for cell division, polarization and differentiation. Here, we found that the CAMSAP C-terminal CKK domain is widely present among eukaryotes and autonomously recognizes microtubule minus ends. Through a combination of structural approaches, we uncovered how mammalian CKK binds between two tubulin dimers at the interprotofilament interface on the outer microtubule surface. In vitro reconstitution assays combined with high-resolution fluorescence microscopy and cryo-electron tomography suggested that CKK preferentially associates with the transition zone between curved protofilaments and the regular microtubule lattice. We propose that minus-end-specific features of the interprotofilament interface at this site serve as the basis for CKK's minus-end preference. The steric clash between microtubule-bound CKK and kinesin motors explains how CKK protects microtubule minus ends against kinesin-13-induced depolymerization and thus controls the stability of free microtubule minus ends. | |||||||||||||||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 715.7 KB | ![]() | |
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Header (meta data) | ![]() ![]() | 17.4 KB 17.4 KB | Display Display | ![]() |
Images | ![]() | 269.5 KB | ||
Filedesc metadata | ![]() | 6.8 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 5m50MC ![]() 3444C ![]() 4156C ![]() 5lznC ![]() 5m54C ![]() 5m5cC 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
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | CAMSAP3 CKK decorated 13pf microtubule asymmetric unit | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.534 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 : Complex of two tubulin dimers with bound CAMSAP3-CKK domain
+Supramolecule #1: Complex of two tubulin dimers with bound CAMSAP3-CKK domain
+Supramolecule #2: tubulin
+Supramolecule #3: CAMSAP3-CKK domain
+Macromolecule #1: Tubulin alpha chain
+Macromolecule #2: Tubulin beta-2B chain
+Macromolecule #3: Calmodulin-regulated spectrin-associated protein 3
+Macromolecule #4: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #5: MAGNESIUM ION
+Macromolecule #6: GUANOSINE-5'-DIPHOSPHATE
+Macromolecule #7: TAXOL
+Macromolecule #8: water
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | filament |
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Sample preparation
Buffer | pH: 6.8 / Details: BRB80 |
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Grid | Model: C-flat-2/2 / Material: COPPER / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE / Instrument: FEI VITROBOT MARK III |
Details | 13pf Microtubules |
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Electron microscopy
Microscope | FEI POLARA 300 |
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Image recording | Film or detector model: DIRECT ELECTRON DE-20 (5k x 3k) / Detector mode: INTEGRATING / Average electron dose: 25.0 e/Å2 / Details: 25e-/A2 used in final reconstuctions |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | ![]() Model: Tecnai Polara / Image courtesy: FEI Company |
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Image processing
Startup model | Type of model: OTHER / Details: Kinesin decorated microtubule |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 5.3 Å / Resolution method: FSC 0.143 CUT-OFF Details: Gold-standard noise substitution test used to asses for over fitting (Chen et al., 2013) Number images used: 6530 |
Initial angle assignment | Type: PROJECTION MATCHING / Software - Name: SPIDER |
Final angle assignment | Type: OTHER / Software - Name: FREALIGN / Details: Projection matching in Fourier space |
-Atomic model buiding 1
Refinement | Protocol: OTHER |
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Output model | ![]() PDB-5m50: |