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- EMDB-60568: Flagellar central pair apparatus of Chlamydomonas reinhardtii wit... -

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Entry
Database: EMDB / ID: EMD-60568
TitleFlagellar central pair apparatus of Chlamydomonas reinhardtii with GFP-tag at the N-terminus of FAP47 protein.
Map dataFlagellar central pair apparatus of Chlamydomonas reinhardii with GFP-tag at the N-terminus of FAP47 protein.
Sample
  • Organelle or cellular component: Flagellar central pair apparatus of Chlamydomonas reinhardtii wild type
Keywordscilia/flagella / Chlamydomonas / MOTOR PROTEIN
Biological speciesChlamydomonas reinhardtii (plant)
Methodsubtomogram averaging / cryo EM / Resolution: 40.2 Å
AuthorsTani Y / Yanagisawa H / Kikkawa M
Funding support Japan, 2 items
OrganizationGrant numberCountry
Japan Society for the Promotion of Science (JSPS)JP21H05248 Japan
Japan Agency for Medical Research and Development (AMED)JP23ama121002 Japan
CitationJournal: Cytoskeleton (Hoboken) / Year: 2024
Title: Structure and function of FAP47 in the central pair apparatus of Chlamydomonas flagella.
Authors: Yuma Tani / Haruaki Yanagisawa / Toshiki Yagi / Masahide Kikkawa /
Abstract: Motile cilia have a so-called "9 + 2" structure, which consists of nine doublet microtubules and a central pair apparatus. The central pair apparatus (CA) is thought to interact mechanically with ...Motile cilia have a so-called "9 + 2" structure, which consists of nine doublet microtubules and a central pair apparatus. The central pair apparatus (CA) is thought to interact mechanically with radial spokes and to control the flagellar beating. Recently, the components of the CA have been identified by proteomic and genomic analyses. Still, the mechanism of how the CA contributes to ciliary motility has much to be revealed. Here, we focused on one CA component with a large molecular weight: FAP47, and its relationship with two other CA components with large molecular weight: HYDIN, and CPC1. The analyses of motility of the Chlamydomonas mutants revealed that in contrast to cpc1 or hydin, which swam more slowly than the wild type, fap47 cells displayed wild-type swimming velocity and flagellar beat frequency, yet interestingly, fap47 cells have phototaxis defects and swim straighter than the wild-type cells. Furthermore, the double mutant fap47cpc1 and fap47hydin showed significantly slower swimming than cpc1 and hydin cells, and the motility defect of fap47cpc1 was rescued to the cpc1 level with GFP-tagged FAP47, indicating that the lack of FAP47 makes the motility defect of cpc1 worse. Cryo-electron tomography demonstrated that the fap47 lacks a part of the C1-C2 bridge of CA. Taken together, these observations indicate that FAP47 maintains the structural stiffness of the CA, which is important for flagellar regulation.
History
DepositionJun 17, 2024-
Header (metadata) releaseJun 26, 2024-
Map releaseJun 26, 2024-
UpdateJul 3, 2024-
Current statusJul 3, 2024Processing site: PDBj / Status: Released

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Structure visualization

Supplemental images

emd_60568.png

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Map

FileDownload / File: emd_60568.map.gz / Format: CCP4 / Size: 30.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationFlagellar central pair apparatus of Chlamydomonas reinhardii with GFP-tag at the N-terminus of FAP47 protein.
Projections & slices

Image control

Size
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AxesZ (Sec.)Y (Row.)X (Col.)
7.1 Å/pix.
x 200 pix.
= 1420. Å		7.1 Å/pix.
x 200 pix.
= 1420. Å		7.1 Å/pix.
x 200 pix.
= 1420. Å

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Images are generated by Spider.

Voxel sizeX=Y=Z: 7.1 Å
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Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 155.0
Minimum - Maximum-1073.571300000000065 - 1215.877300000000105
Average (Standard dev.)-7.1840386 (±98.236519999999999)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions200200200
Spacing200200200
CellA=B=C: 1420.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #1

Fileemd_60568_half_map_1.map
Projections & Slices
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Half map: #2

Fileemd_60568_half_map_2.map
Projections & Slices
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Histogram (log scale)

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Sample components

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Entire : Flagellar central pair apparatus of Chlamydomonas reinhardtii wil...

EntireName: Flagellar central pair apparatus of Chlamydomonas reinhardtii wild type
Components
  • Organelle or cellular component: Flagellar central pair apparatus of Chlamydomonas reinhardtii wild type

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Supramolecule #1: Flagellar central pair apparatus of Chlamydomonas reinhardtii wil...

SupramoleculeName: Flagellar central pair apparatus of Chlamydomonas reinhardtii wild type
type: organelle_or_cellular_component / ID: 1 / Parent: 0
Source (natural)Organism: Chlamydomonas reinhardtii (plant)

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Experimental details

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Structure determination

Methodcryo EM
Processingsubtomogram averaging
Aggregation statefilament

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Sample preparation

BufferpH: 7.3
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TALOS ARCTICA
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 3.2 e/Å2
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 8.0 µm / Nominal defocus min: 6.0 µm
Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company

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Image processing

Final reconstructionResolution.type: BY AUTHOR / Resolution: 40.2 Å / Resolution method: FSC 0.5 CUT-OFF / Number subtomograms used: 2400
ExtractionNumber tomograms: 52 / Number images used: 2643
Final angle assignmentType: NOT APPLICABLE
FSC plot (resolution estimation)

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