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- EMDB-4267: Arp2/3 complex in open conformation -

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Basic information

Entry
Database: EMDB / ID: EMD-4267
TitleArp2/3 complex in open conformation
Map data
SampleArp2/3 complex with bound Abp1 dimer
Biological speciesSaccharomyces cerevisiae (baker's yeast)
Methodsingle particle reconstruction / Resolution: 18 Å
AuthorsSokolova OS / Goode BL
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of HealthR01-GM063691 United States
CitationJournal: Nat Commun / Year: 2018
Title: Abp1 promotes Arp2/3 complex-dependent actin nucleation and stabilizes branch junctions by antagonizing GMF.
Authors: Siyang Guo / Olga S Sokolova / Johnson Chung / Shae Padrick / Jeff Gelles / Bruce L Goode /
Abstract: Formation and turnover of branched actin networks underlies cell migration and other essential force-driven processes. Type I nucleation-promoting factors (NPFs) such as WASP recruit actin monomers ...Formation and turnover of branched actin networks underlies cell migration and other essential force-driven processes. Type I nucleation-promoting factors (NPFs) such as WASP recruit actin monomers to Arp2/3 complex to stimulate nucleation. In contrast, mechanisms of type II NPFs such as Abp1 (also known as HIP55 and Drebrin-like protein) are less well understood. Here, we use single-molecule analysis to investigate yeast Abp1 effects on Arp2/3 complex, and find that Abp1 strongly enhances Arp2/3-dependent branch nucleation by stabilizing Arp2/3 on sides of mother filaments. Abp1 binds dynamically to filament sides, with sub-second lifetimes, yet associates stably with branch junctions. Further, we uncover a role for Abp1 in protecting filament junctions from GMF-induced debranching by competing with GMF for Arp2/3 binding. These data, combined with EM structures of Abp1 dimers bound to Arp2/3 complex in two different conformations, expand our mechanistic understanding of type II NPFs.
History
DepositionJan 23, 2018-
Header (metadata) releaseMar 14, 2018-
Map releaseMar 20, 2019-
UpdateOct 2, 2019-
Current statusOct 2, 2019Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.18
  • Imaged by UCSF Chimera
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  • Surface view colored by radius
  • Surface level: 0.18
  • Imaged by UCSF Chimera
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Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

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Map

FileDownload / File: emd_4267.map.gz / Format: CCP4 / Size: 3.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
2.5 Å/pix.
x 100 pix.
= 250. Å
2.5 Å/pix.
x 100 pix.
= 250. Å
2.5 Å/pix.
x 100 pix.
= 250. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 2.5 Å
Density
Contour LevelBy AUTHOR: 0.18 / Movie #1: 0.18
Minimum - Maximum-0.6249865 - 1.9891132
Average (Standard dev.)0.017773548 (±0.12210158)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin-50-50-50
Dimensions100100100
Spacing100100100
CellA=B=C: 250.0 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z2.52.52.5
M x/y/z100100100
origin x/y/z0.0000.0000.000
length x/y/z250.000250.000250.000
α/β/γ90.00090.00090.000
start NX/NY/NZ000
NX/NY/NZ320320320
MAP C/R/S123
start NC/NR/NS-50-50-50
NC/NR/NS100100100
D min/max/mean-0.6251.9890.018

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Supplemental data

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Sample components

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Entire Arp2/3 complex with bound Abp1 dimer

EntireName: Arp2/3 complex with bound Abp1 dimer / Number of components: 1

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Component #1: cellular-component, Arp2/3 complex with bound Abp1 dimer

Cellular-componentName: Arp2/3 complex with bound Abp1 dimer / Recombinant expression: No
SourceSpecies: Saccharomyces cerevisiae (baker's yeast)

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Experimental details

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Sample preparation

SpecimenSpecimen state: Particle
Sample solutionpH: 7.5
VitrificationCryogen name: NONE

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Electron microscopy imaging

ImagingMicroscope: JEOL 2100
Electron gunElectron source: LAB6 / Accelerating voltage: 200 kV / Electron dose: 13 e/Å2 / Illumination mode: FLOOD BEAM
LensImaging mode: BRIGHT FIELD
Specimen HolderModel: OTHER
CameraDetector: GATAN ULTRASCAN 1000 (2k x 2k)

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Image processing

ProcessingMethod: single particle reconstruction / Number of projections: 2500
3D reconstructionResolution: 18 Å / Resolution method: FSC 0.5 CUT-OFF

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