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- EMDB-47150: Mitochondrial fission site in neurons at the constriction stage -

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Basic information

Entry
Database: EMDB / ID: EMD-47150
TitleMitochondrial fission site in neurons at the constriction stage
Map data
Sample
  • Cell: Rat hippocampal neurons
KeywordsFMRP-RNA granule / local translation / neuron / mitochondrial fission / RIBOSOME
Biological speciesRattus norvegicus (Norway rat)
Methodelectron tomography / cryo EM
AuthorsPeng R / Chang Y-W
Funding support United States, 1 items
OrganizationGrant numberCountry
David and Lucile Packard Foundation2019-69645 United States
CitationJournal: Nat Cell Biol / Year: 2024
Title: FMRP regulates MFF translation to locally direct mitochondrial fission in neurons.
Authors: Adam R Fenton / Ruchao Peng / Charles Bond / Siewert Hugelier / Melike Lakadamyali / Yi-Wei Chang / Erika L F Holzbaur / Thomas A Jongens /
Abstract: Fragile X messenger ribonucleoprotein (FMRP) is a critical regulator of translation, whose dysfunction causes fragile X syndrome. FMRP dysfunction disrupts mitochondrial health in neurons, but it is ...Fragile X messenger ribonucleoprotein (FMRP) is a critical regulator of translation, whose dysfunction causes fragile X syndrome. FMRP dysfunction disrupts mitochondrial health in neurons, but it is unclear how FMRP supports mitochondrial homoeostasis. Here we demonstrate that FMRP granules are recruited to the mitochondrial midzone, where they mark mitochondrial fission sites in axons and dendrites. Endolysosomal vesicles contribute to FMRP granule positioning around mitochondria and facilitate FMRP-associated fission via Rab7 GTP hydrolysis. Cryo-electron tomography and real-time translation imaging reveal that mitochondria-associated FMRP granules are ribosome-rich structures that serve as sites of local protein synthesis. Specifically, FMRP promotes local translation of mitochondrial fission factor (MFF), selectively enabling replicative fission at the mitochondrial midzone. Disrupting FMRP function dysregulates mitochondria-associated MFF translation and perturbs fission dynamics, resulting in increased peripheral fission and an irregular distribution of mitochondrial nucleoids. Thus, FMRP regulates local translation of MFF in neurons, enabling precise control of mitochondrial fission.
History
DepositionSep 30, 2024-
Header (metadata) releaseOct 16, 2024-
Map releaseOct 16, 2024-
UpdateJan 22, 2025-
Current statusJan 22, 2025Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_47150.png

Downloads & links

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Map

FileDownload / File: emd_47150.map.gz / Format: CCP4 / Size: 1.3 GB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
10.6 Å/pix.
x 240 pix.
= 2544. Å		10.6 Å/pix.
x 1440 pix.
= 15264.001 Å		10.6 Å/pix.
x 1022 pix.
= 10833.2 Å

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

generated in cubic-lattice coordinate

Voxel sizeX=Y=Z: 10.6 Å
Density

Histogram

Histogram (log scale)
Minimum - Maximum-1637858360000000000.0 - 1098834190000000000.0
Average (Standard dev.)-646772080000000.0 (±61967810000000000.0)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin00313
Dimensions14401022240
Spacing10221440240
CellA: 10833.2 Å / B: 15264.001 Å / C: 2544.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Rat hippocampal neurons

EntireName: Rat hippocampal neurons
Components
  • Cell: Rat hippocampal neurons

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Supramolecule #1: Rat hippocampal neurons

SupramoleculeName: Rat hippocampal neurons / type: cell / ID: 1 / Parent: 0
Details: Rat hippocampal neurons were cultured on cryo-EM grids and transfected with plasmids encoding EGFP-FMRP.
Source (natural)Organism: Rattus norvegicus (Norway rat)

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Experimental details

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Structure determination

Methodcryo EM
Processingelectron tomography
Aggregation statecell

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Sample preparation

BufferpH: 7.4
GridModel: Quantifoil R2/2 / Material: GOLD / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec. / Pretreatment - Atmosphere: AIR
VitrificationCryogen name: ETHANE-PROPANE / Chamber humidity: 95 % / Chamber temperature: 310 K / Instrument: LEICA EM GP
SectioningOther: NO SECTIONING
Fiducial markerManufacturer: BBI Solutions / Diameter: 10 nm

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Electron microscopy

MicroscopeTFS KRIOS
Specialist opticsPhase plate: VOLTA PHASE PLATE / Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 2.65 e/Å2 / Details: total dose per tilt series = 105 e/A2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.0 µm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionAlgorithm: BACK PROJECTION / Software - Name: IMOD (ver. 4.11) / Number images used: 41

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