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- EMDB-16877: Subtomogram averaging structure of cofilactin filament inside mic... -
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Open data
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Basic information
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Title | Subtomogram averaging structure of cofilactin filament inside microtubule lumen of Drosophila S2 cell protrusion. | |||||||||
![]() | Masked map at 16.5 A resolution. Sharpened with a B-factor of -500 A2. | |||||||||
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![]() | Cytoskeleton / Filament / Actin / Cofilactin / CONTRACTILE PROTEIN | |||||||||
Function / homology | ![]() establishment of imaginal disc-derived wing hair orientation / establishment of ommatidial planar polarity / imaginal disc-derived leg segmentation / meiotic cytokinesis / Gap junction degradation / Formation of annular gap junctions / EPHB-mediated forward signaling / EPH-ephrin mediated repulsion of cells / VEGFA-VEGFR2 Pathway / Cell-extracellular matrix interactions ...establishment of imaginal disc-derived wing hair orientation / establishment of ommatidial planar polarity / imaginal disc-derived leg segmentation / meiotic cytokinesis / Gap junction degradation / Formation of annular gap junctions / EPHB-mediated forward signaling / EPH-ephrin mediated repulsion of cells / VEGFA-VEGFR2 Pathway / Cell-extracellular matrix interactions / RHOBTB2 GTPase cycle / RHOF GTPase cycle / MAP2K and MAPK activation / Platelet degranulation / RHO GTPases Activate WASPs and WAVEs / Regulation of actin dynamics for phagocytic cup formation / compound eye morphogenesis / DNA Damage Recognition in GG-NER / mushroom body development / rhabdomere development / actomyosin contractile ring assembly / actin filament fragmentation / Clathrin-mediated endocytosis / border follicle cell migration / compound eye development / UCH proteinases / sperm individualization / establishment of planar polarity / centrosome separation / epithelial structure maintenance / maintenance of protein location in cell / brahma complex / actin filament severing / actin filament depolymerization / Ino80 complex / tube formation / regulation of lamellipodium assembly / lamellipodium assembly / female gonad development / mitotic cytokinesis / actin filament polymerization / axonogenesis / actin filament organization / positive regulation of protein secretion / Hydrolases; Acting on acid anhydrides; Acting on acid anhydrides to facilitate cellular and subcellular movement / nuclear matrix / actin filament binding / actin cytoskeleton / actin binding / cytoskeleton / hydrolase activity / chromatin remodeling / nucleoplasm / ATP binding / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 16.5 Å | |||||||||
![]() | Ventura Santos C / Carter AP | |||||||||
Funding support | ![]()
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![]() | ![]() Title: CryoET shows cofilactin filaments inside the microtubule lumen. Authors: Camilla Ventura Santos / Stephen L Rogers / Andrew P Carter / ![]() Abstract: Cytoplasmic microtubules are tubular polymers that can harbor small proteins or filaments inside their lumen. The identity of these objects and what causes their accumulation has not been ...Cytoplasmic microtubules are tubular polymers that can harbor small proteins or filaments inside their lumen. The identity of these objects and what causes their accumulation has not been conclusively established. Here, we used cryogenic electron tomography (cryoET) of S2 cell protrusions and found filaments inside the microtubule lumen, which resemble those reported recently in human HAP1 cells. The frequency of these filaments increased upon inhibition of the sarco/endoplasmic reticulum Ca ATPase (SERCA) with the small-molecule drug thapsigargin. Subtomogram averaging showed that the luminal filaments adopt a helical structure reminiscent of cofilin-bound actin (cofilactin). Consistent with this, cofilin was activated in cells under the same conditions that increased luminal filament occurrence. Furthermore, RNAi knock-down of cofilin reduced the frequency of luminal filaments with cofilactin morphology. These results suggest that cofilin activation stimulates its accumulation on actin filaments inside the microtubule lumen. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 5.1 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 19.2 KB 19.2 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 9.2 KB | Display | ![]() |
Images | ![]() | 23.4 KB | ||
Masks | ![]() | 64 MB | ![]() | |
Filedesc metadata | ![]() | 6.2 KB | ||
Others | ![]() ![]() ![]() | 59.6 MB 49.6 MB 49.6 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8oh4MC 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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Related items in Molecule of the Month |
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Map
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Annotation | Masked map at 16.5 A resolution. Sharpened with a B-factor of -500 A2. | ||||||||||||||||||||
Voxel size | X=Y=Z: 2.83 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
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Density Histograms |
-Additional map: Unmasked map at 16.5 A resolution. Sharpened with...
File | emd_16877_additional_1.map | ||||||||||||
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Annotation | Unmasked map at 16.5 A resolution. Sharpened with a B-factor of -500 A2. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 2.
File | emd_16877_half_map_1.map | ||||||||||||
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Annotation | Half map 2. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 1.
File | emd_16877_half_map_2.map | ||||||||||||
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Annotation | Half map 1. | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Cofilactin filament inside the microtubule lumen of induced Droso...
Entire | Name: Cofilactin filament inside the microtubule lumen of induced Drosophila S2 cell protrusion |
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Components |
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-Supramolecule #1: Cofilactin filament inside the microtubule lumen of induced Droso...
Supramolecule | Name: Cofilactin filament inside the microtubule lumen of induced Drosophila S2 cell protrusion type: cell / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() ![]() |
-Macromolecule #1: Actin-5C
Macromolecule | Name: Actin-5C / type: protein_or_peptide / ID: 1 / Number of copies: 8 / Enantiomer: LEVO EC number: Hydrolases; Acting on acid anhydrides; Acting on acid anhydrides to facilitate cellular and subcellular movement |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 41.16098 KDa |
Sequence | String: AALVVDNGSG MCKAGFAGDD APRAVFPSIV GRPRHQGVMV GMGQKDSYVG DEAQSKRGIL TLKYPIEHGI VTNWDDMEKI WHHTFYNEL RVAPEEHPVL LTEAPLNPKA NREKMTQIMF ETFNTPAMYV AIQAVLSLYA SGRTTGIVLD SGDGVSHTVP I YEGYALPH ...String: AALVVDNGSG MCKAGFAGDD APRAVFPSIV GRPRHQGVMV GMGQKDSYVG DEAQSKRGIL TLKYPIEHGI VTNWDDMEKI WHHTFYNEL RVAPEEHPVL LTEAPLNPKA NREKMTQIMF ETFNTPAMYV AIQAVLSLYA SGRTTGIVLD SGDGVSHTVP I YEGYALPH AILRLDLAGR DLTDYLMKIL TERGYSFTTT AEREIVRDIK EKLCYVALDF EQEMATAASS SSLEKSYELP DG QVITIGN ERFRCPEALF QPSFLGMEAC GIHETTYNSI MKCDVDIRKD LYANTVLSGG TTMYPGIADR MQKEITALAP STM KIKIIA PPERKYSVWI GGSILASLST FQQMWISKQE YDESGPSIVH RKCF UniProtKB: Actin-5C |
-Macromolecule #2: Cofilin/actin-depolymerizing factor homolog
Macromolecule | Name: Cofilin/actin-depolymerizing factor homolog / type: protein_or_peptide / ID: 2 / Number of copies: 6 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 17.180529 KDa |
Sequence | String: MASGVTVSDV CKTTYEEIKK DKKHRYVIFY IRDEKQIDVE TVADRNAEYD QFLEDIQKCG PGECRYGLFD FEYMHQCQGT SESSKKQKL FLMSWCPDTA KVKKKMLYSS SFDALKKSLV GVQKYIQATD LSEASREAVE EKLRATDRQ UniProtKB: Cofilin/actin-depolymerizing factor homolog |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | subtomogram averaging |
Aggregation state | filament |
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Sample preparation
Buffer | pH: 7 |
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Grid | Model: Quantifoil R3.5/1 / Material: GOLD / Mesh: 200 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. Details: Quantifoil R3.5/1 Au200 grids were glow discharged for 30s at 20 - 30 mA and subsequently coated with 0.25 ug/mL Concanavalin Aater for 1 - 16 h at 37 degrees Celcius. |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 198.15 K / Instrument: FEI VITROBOT MARK III |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 3.0 e/Å2 Details: Data was collected on Gatan K2 summit (2.952 A/pixel) and Gatan K3 summit (2.659 A/pixel) with 3 degree increments (3 e/A2 dose per tilt). The total dose was between 118 and 122 e/A2. |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 6.0 µm / Nominal defocus min: 2.5 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Details | Drosophila cofilin and actin structures were predicted as a complex with Alphafold 2-Multimer. Actin from the predicted cofilin-actin complex was iteratively aligned with 8 actin subunits in the chicken cofilactin PDB model 5yU8. Two cofilin moieties at the pointed end were removed. Side chains were truncated. The model was not refined. |
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Output model | ![]() PDB-8oh4: |