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- EMDB-10760: Mammalian 48S late-stage initiation complex with beta-globin mRNA -

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Basic information

Entry
Database: EMDB / ID: EMD-10760
TitleMammalian 48S late-stage initiation complex with beta-globin mRNA
Map data
SampleLS48S IC with beta-globin mRNA
  • LS48S IC
  • beta-globin mRNA
  • (nucleic-acidNucleic acid) x 3
  • 60s ribosomal protein ul41
  • (40S Ribosomal protein ...) x 32
  • ribosomal protein RACK1
  • (Eukaryotic translation initiation factor 2 subunit ...) x 2
  • eukaryotic translation initiation factor 1A
  • ATP-binding cassette sub-family E member 1 (ABCE1)
  • (ligand) x 3
Function / homology
Function and homology information


laminin receptor activity / mammalian oogenesis stage / activation-induced cell death of T cells / DNA-(apurinic or apyrimidinic site) lyase / class I DNA-(apurinic or apyrimidinic site) endonuclease activity / erythrocyte development / TOR signaling / regulation of translational fidelity / T cell proliferation involved in immune response / mitotic cell cycle checkpoint ...laminin receptor activity / mammalian oogenesis stage / activation-induced cell death of T cells / DNA-(apurinic or apyrimidinic site) lyase / class I DNA-(apurinic or apyrimidinic site) endonuclease activity / erythrocyte development / TOR signaling / regulation of translational fidelity / T cell proliferation involved in immune response / mitotic cell cycle checkpoint / ribosomal small subunit biogenesis / gastrulation / polysome / laminin binding / translation initiation factor activity / polysomal ribosome / ribosomal small subunit assembly / cytoplasmic ribonucleoprotein granule / rough endoplasmic reticulum / placenta development / rRNA processing / cytoplasmic stress granule / spindle / cytoplasmic translation / G1/S transition of mitotic cell cycle / cell body / cytosolic small ribosomal subunit / regulation of translation / small ribosomal subunit / glucose homeostasis / T cell differentiation in thymus / mitochondrial inner membrane / rRNA binding / ribosome / structural constituent of ribosome / translation / cell differentiation / cell cycle / ATPase activity / cell division / mRNA binding / DNA repair / positive regulation of apoptotic process / dendrite / apoptotic process / nucleolus / positive regulation of cell population proliferation / protein kinase binding / negative regulation of apoptotic process / perinuclear region of cytoplasm / endoplasmic reticulum / RNA binding / zinc ion binding / ATP binding / plasma membrane / metal ion binding / nucleus / cytosol / cytoplasm
Ribosomal protein L23/L15e core domain superfamily / Ribosomal protein S3, eukaryotic/archaeal / Ribosomal protein L7Ae/L30e/S12e/Gadd45 / K Homology domain, type 2 / Ribosomal protein S25 / Ribosomal protein S5, C-terminal / Plectin/S10, N-terminal / Ribosomal protein S23, eukaryotic/archaeal / Ribosomal protein S2, eukaryotic/archaeal / ABC transporter-like ...Ribosomal protein L23/L15e core domain superfamily / Ribosomal protein S3, eukaryotic/archaeal / Ribosomal protein L7Ae/L30e/S12e/Gadd45 / K Homology domain, type 2 / Ribosomal protein S25 / Ribosomal protein S5, C-terminal / Plectin/S10, N-terminal / Ribosomal protein S23, eukaryotic/archaeal / Ribosomal protein S2, eukaryotic/archaeal / ABC transporter-like / Ribosomal protein S4/S9, eukaryotic/archaeal / Ribosomal protein S5, eukaryotic/archaeal / Ribosomal protein S19A/S15e / Ribosomal protein S5/S7, eukaryotic/archaeal / Ribosomal protein S10, eukaryotic/archaeal / KOW / AAA+ ATPase domain / S1 domain / RNA-binding domain, S1, IF1 type / Ribosomal protein S2 / Ribosomal protein S19e / Ribosomal protein S3, C-terminal / Ribosomal protein S6e / Ribosomal protein S3Ae / WD40 repeat / Ribosomal protein S10 / Ribosomal protein S13 / RNA-binding S4 domain / Ribosomal protein S4/S9, N-terminal / Ribosomal protein S21e / Ribosomal protein S11 / Ribosomal protein S24e / Ribosomal protein S19/S15 / Ribosomal protein S27a / Ribosomal protein S12/S23 / Ribosomal protein S30 / Ribosomal protein S17e / Translation initiation factor 1A (eIF-1A), conserved site / 4Fe-4S ferredoxin-type, iron-sulphur binding domain / 4Fe-4S ferredoxin, iron-sulphur binding, conserved site / WD40-repeat-containing domain / Ribosomal protein S4, conserved site / Ribosomal S24e conserved site / Ribosomal S11, conserved site / Ribosomal protein S2, conserved site / K homology domain-like, alpha/beta / Ribosomal protein S4e, N-terminal, conserved site / Ribosomal protein S10, conserved site / Ribosomal protein S13, conserved site / Ribosomal protein S14, conserved site / Ribosomal protein S17e, conserved site / Ribosomal protein S19e, conserved site / ABC transporter, conserved site / WD40/YVTN repeat-like-containing domain superfamily / RNase L inhibitor RLI-like, possible metal-binding domain / Nucleic acid-binding, OB-fold / Ribosomal protein L41 / K homology domain superfamily, prokaryotic type / S15/NS1, RNA-binding / Ribosomal protein S13-like, H2TH / Zinc-binding ribosomal protein / Translation initiation factor 2, alpha subunit / Ribosomal protein S13/S15, N-terminal / Ribosomal protein L2, domain 2 / Ribosomal protein S3Ae, conserved site / RLI1 / Ribosomal protein S5, N-terminal / Ribosomal protein S4e, N-terminal / Ribosomal protein S4e, central region / Ribosomal protein S6, eukaryotic / Ribosomal protein S5 domain 2-type fold, subgroup / Translation initiation factor 1A (eIF-1A) / Ribosomal protein S14 / Ribosomal protein S3, conserved site / Ubiquitin-like domain superfamily / WD40-repeat-containing domain superfamily / Ribosomal protein S8 superfamily / RLI, domain 1 / 40S ribosomal protein S11, N-terminal / 40S ribosomal protein SA, C-terminal domain / 40S ribosomal protein S4, C-terminal domain / 50S ribosomal protein L30e-like / Winged helix DNA-binding domain superfamily / Ribosomal protein S28e conserved site / Ribosomal protein S17, archaeal/eukaryotic / 40S ribosomal protein SA / 40S ribosomal protein S1/3, eukaryotes / Ribosomal protein S2, eukaryotic / Ribosomal protein S10 domain / Winged helix-like DNA-binding domain superfamily / Ribosomal protein S17e-like superfamily / Ribosomal protein S8e / Ribosomal protein S26e superfamily / Ribosomal protein S14/S29 / Ribosomal protein S8e subdomain, eukaryotes / Ribosomal protein S4, KOW domain / 40S ribosomal protein S29/30S ribosomal protein S14 type Z / S27a-like superfamily / Ribosomal protein S21e superfamily / Ribosomal protein S4e, central domain superfamily
40S ribosomal protein S3 / 40S ribosomal protein S4 / 40S ribosomal protein SA / 40S ribosomal protein S6 / 40S ribosomal protein S21 / 40S ribosomal protein S19 / 40S ribosomal protein S3a / 40S ribosomal protein S11 / 40S ribosomal protein S18 / S10_plectin domain-containing protein ...40S ribosomal protein S3 / 40S ribosomal protein S4 / 40S ribosomal protein SA / 40S ribosomal protein S6 / 40S ribosomal protein S21 / 40S ribosomal protein S19 / 40S ribosomal protein S3a / 40S ribosomal protein S11 / 40S ribosomal protein S18 / S10_plectin domain-containing protein / 40S ribosomal protein S28 / 40S ribosomal protein S24 / 40S ribosomal protein S17 / 40S ribosomal protein S2 / 40S ribosomal protein S27 / 40S ribosomal protein S15 / 40S ribosomal protein S29 / 40S ribosomal protein S8 / 40S ribosomal protein S25 / 40S ribosomal protein S15a / WD_REPEATS_REGION domain-containing protein / 60S ribosomal protein L41 / 40S ribosomal protein S6 / 40S ribosomal protein S9 / 40S ribosomal protein S12 / Uncharacterized protein / Uncharacterized protein / 40S ribosomal protein S20 / 40S ribosomal protein S27a / 40S ribosomal protein S26 / 40S ribosomal protein S13 / 40S ribosomal protein S7 / Eukaryotic translation initiation factor 4C / 40S ribosomal protein S23 / Uncharacterized protein / Eukaryotic translation initiation factor 2 subunit 1 / 40S ribosomal protein S30 / Ribosomal protein S5
Biological speciesOryctolagus cuniculus (rabbit) / Homo sapiens (human) / Rabbit (rabbit)
Methodsingle particle reconstruction / cryo EM / Resolution: 3 Å
AuthorsBochler A / Simonetti A / Guca E / Hashem Y
Funding supportEuropean Union, France, 2 items
OrganizationGrant numberCountry
European Research Council (ERC)759120European Union
French National Research AgencyANR-14-ACHN-0024 France
CitationJournal: Cell Rep / Year: 2020
Title: Structural Insights into the Mammalian Late-Stage Initiation Complexes.
Authors: Angelita Simonetti / Ewelina Guca / Anthony Bochler / Lauriane Kuhn / Yaser Hashem /
Abstract: In higher eukaryotes, the mRNA sequence in the direct vicinity of the start codon, called the Kozak sequence (CRCCaugG, where R is a purine), is known to influence the rate of the initiation process. ...In higher eukaryotes, the mRNA sequence in the direct vicinity of the start codon, called the Kozak sequence (CRCCaugG, where R is a purine), is known to influence the rate of the initiation process. However, the molecular basis underlying its role remains poorly understood. Here, we present the cryoelectron microscopy (cryo-EM) structures of mammalian late-stage 48S initiation complexes (LS48S ICs) in the presence of two different native mRNA sequences, β-globin and histone 4, at overall resolution of 3 and 3.5 Å, respectively. Our high-resolution structures unravel key interactions from the mRNA to eukaryotic initiation factors (eIFs): 1A, 2, 3, 18S rRNA, and several 40S ribosomal proteins. In addition, we are able to study the structural role of ABCE1 in the formation of native 48S ICs. Our results reveal a comprehensive map of ribosome/eIF-mRNA and ribosome/eIF-tRNA interactions and suggest the impact of mRNA sequence on the structure of the LS48S IC.
Validation ReportSummary, Full report, XML, About validation report
History
DepositionMar 12, 2020-
Header (metadata) releaseApr 8, 2020-
Map releaseApr 8, 2020-
UpdateApr 22, 2020-
Current statusApr 22, 2020Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.02
  • Imaged by UCSF Chimera
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  • Surface view colored by height
  • Surface level: 0.02
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-6yal
  • Surface level: 0.02
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

FileDownload / File: emd_10760.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.1 Å/pix.
x 384 pix.
= 422.4 Å
1.1 Å/pix.
x 384 pix.
= 422.4 Å
1.1 Å/pix.
x 384 pix.
= 422.4 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.1 Å
Density
Contour LevelBy AUTHOR: 0.02 / Movie #1: 0.02
Minimum - Maximum-0.07795042 - 0.1473876
Average (Standard dev.)0.00010984154 (±0.0057947584)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions384384384
Spacing384384384
CellA=B=C: 422.40002 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.11.11.1
M x/y/z384384384
origin x/y/z0.0000.0000.000
length x/y/z422.400422.400422.400
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS384384384
D min/max/mean-0.0780.1470.000

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Supplemental data

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Sample components

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Entire LS48S IC with beta-globin mRNA

EntireName: LS48S IC with beta-globin mRNA / Number of components: 47

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Component #1: protein, LS48S IC with beta-globin mRNA

ProteinName: LS48S IC with beta-globin mRNA / Recombinant expression: No

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Component #2: protein, LS48S IC

ProteinName: LS48S IC / Recombinant expression: No
SourceSpecies: Oryctolagus cuniculus (rabbit)

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Component #3: protein, beta-globin mRNA

ProteinName: beta-globin mRNA / Recombinant expression: No
SourceSpecies: Homo sapiens (human)
Source (engineered)Expression System: synthetic construct (others)

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Component #4: nucleic-acid, initiator methionylated tRNA

nucleic acidName: initiator methionylated tRNA / Class: RNA
Details: A at position 37 is modified (threonyl-carbamoyl-adenosine (t6A))
Structure: OTHER / Synthetic: No
Sequence:
AGCAGAGUGG CGCAGCGGAA GCGUGCUGGG CCCAU(T6A)ACCC AGAGGUCGAU GGAUCGAAAC CAUCCUCUGC UACCA
MassTheoretical: 24.376625 kDa
SourceSpecies: Rabbit (rabbit)

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Component #5: protein, 60s ribosomal protein ul41

ProteinName: 60s ribosomal protein ul41 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 3.473451 kDa
SourceSpecies: Rabbit (rabbit)

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Component #6: protein, 40S ribosomal protein uS2

ProteinName: 40S ribosomal protein uS2 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 23.491988 kDa
SourceSpecies: Rabbit (rabbit)

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Component #7: protein, 40S ribosomal protein eS1

ProteinName: 40S ribosomal protein eS1 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 29.928967 kDa
SourceSpecies: Rabbit (rabbit)

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Component #8: protein, 40S ribosomal protein uS5

ProteinName: 40S ribosomal protein uS5 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 24.814285 kDa
SourceSpecies: Rabbit (rabbit)

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Component #9: protein, 40S Ribosomal protein uS3

ProteinName: 40S Ribosomal protein uS3 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 26.715344 kDa
SourceSpecies: Rabbit (rabbit)

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Component #10: protein, 40S ribosomal protein eS4

ProteinName: 40S ribosomal protein eS4 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 29.65892 kDa
SourceSpecies: Rabbit (rabbit)

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Component #11: protein, 40S Ribosomal protein uS7

ProteinName: 40S Ribosomal protein uS7 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 21.525941 kDa
SourceSpecies: Rabbit (rabbit)

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Component #12: protein, 40S ribosomal protein eS6

ProteinName: 40S ribosomal protein eS6 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 27.471535 kDa
SourceSpecies: Rabbit (rabbit)

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Component #13: protein, 40S ribosomal protein eS7

ProteinName: 40S ribosomal protein eS7 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 21.890545 kDa
SourceSpecies: Rabbit (rabbit)

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Component #14: protein, 40S ribosomal protein eS8

ProteinName: 40S ribosomal protein eS8 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 23.902871 kDa
SourceSpecies: Rabbit (rabbit)

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Component #15: protein, 40S ribosomal protein uS4

ProteinName: 40S ribosomal protein uS4 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 21.266166 kDa
SourceSpecies: Rabbit (rabbit)

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Component #16: protein, 40S ribosomal protein eS10

ProteinName: 40S ribosomal protein eS10 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 11.773953 kDa
SourceSpecies: Rabbit (rabbit)

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Component #17: protein, 40S ribosomal protein uS17

ProteinName: 40S ribosomal protein uS17 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 18.468826 kDa
SourceSpecies: Rabbit (rabbit)

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Component #18: protein, 40S ribosomal protein eS12

ProteinName: 40S ribosomal protein eS12 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 14.538987 kDa
SourceSpecies: Rabbit (rabbit)

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Component #19: protein, 40S ribosomal protein uS15

ProteinName: 40S ribosomal protein uS15 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 17.128191 kDa
SourceSpecies: Rabbit (rabbit)

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Component #20: protein, 40S ribosomal protein uS11

ProteinName: 40S ribosomal protein uS11 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 16.302772 kDa
SourceSpecies: Rabbit (rabbit)

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Component #21: protein, 40S ribosomal protein uS9

ProteinName: 40S ribosomal protein uS9 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 15.975753 kDa
SourceSpecies: Rabbit (rabbit)

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Component #22: protein, 40S ribosomal protein eS17

ProteinName: 40S ribosomal protein eS17 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 15.552119 kDa
SourceSpecies: Rabbit (rabbit)

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Component #23: protein, 40S ribosomal protein eS19

ProteinName: 40S ribosomal protein eS19 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 15.812286 kDa
SourceSpecies: Rabbit (rabbit)

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Component #24: protein, 40S ribosomal protein uS10

ProteinName: 40S ribosomal protein uS10 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 13.398763 kDa
SourceSpecies: Rabbit (rabbit)

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Component #25: protein, 40S ribosomal protein eS21

ProteinName: 40S ribosomal protein eS21 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 8.896102 kDa
SourceSpecies: Rabbit (rabbit)

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Component #26: protein, 40S ribosomal protein uS8

ProteinName: 40S ribosomal protein uS8 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 14.865555 kDa
SourceSpecies: Rabbit (rabbit)

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Component #27: protein, 40S ribosomal protein uS12

ProteinName: 40S ribosomal protein uS12 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 15.844666 kDa
SourceSpecies: Rabbit (rabbit)

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Component #28: protein, 40S ribosomal protein eS24

ProteinName: 40S ribosomal protein eS24 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 15.434267 kDa
SourceSpecies: Rabbit (rabbit)

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Component #29: protein, 40S ribosomal protein eS26

ProteinName: 40S ribosomal protein eS26 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 12.961455 kDa
SourceSpecies: Rabbit (rabbit)

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Component #30: protein, 40S ribosomal protein eS27

ProteinName: 40S ribosomal protein eS27 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 9.480186 kDa
SourceSpecies: Rabbit (rabbit)

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Component #31: protein, 40S ribosomal protein eS28

ProteinName: 40S ribosomal protein eS28 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 7.855052 kDa
SourceSpecies: Rabbit (rabbit)

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Component #32: protein, 40S ribosomal protein uS14

ProteinName: 40S ribosomal protein uS14 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 6.690821 kDa
SourceSpecies: Rabbit (rabbit)

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Component #33: protein, 40S ribosomal protein eS31

ProteinName: 40S ribosomal protein eS31 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 8.358903 kDa
SourceSpecies: Rabbit (rabbit)

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Component #34: protein, ribosomal protein RACK1

ProteinName: ribosomal protein RACK1 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 34.669113 kDa
SourceSpecies: Rabbit (rabbit)

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Component #35: protein, 40S ribosomal protein eS25

ProteinName: 40S ribosomal protein eS25 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 13.645028 kDa
SourceSpecies: Rabbit (rabbit)

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Component #36: protein, 40S ribosomal protein eS30

ProteinName: 40S ribosomal protein eS30 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 14.498884 kDa
SourceSpecies: Rabbit (rabbit)

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Component #37: nucleic-acid, 18S ribosomal RNA

nucleic acidName: 18S ribosomal RNA / Class: RNA / Details: U at position 1244 is modified / Structure: OTHER / Synthetic: No
Sequence: UACCUGGUUG AUCCUGCCAG UAGCAUAUGC UUGUCUCAAA GAUUAAGCCA UGCAUGUCUA AGUACGCACG GCCGGUACAG UGAAACUGCG AAUGGCUCAU UAAAUCAGUU AUGGUUCCUU UGGUCGCUCG CUCCUCUCCU ACUUGGAUAA CUGUGGUAAU UCUAGAGCUA ...Sequence:
UACCUGGUUG AUCCUGCCAG UAGCAUAUGC UUGUCUCAAA GAUUAAGCCA UGCAUGUCUA AGUACGCACG GCCGGUACAG UGAAACUGCG AAUGGCUCAU UAAAUCAGUU AUGGUUCCUU UGGUCGCUCG CUCCUCUCCU ACUUGGAUAA CUGUGGUAAU UCUAGAGCUA AUACAUGCCG ACGGCGCUGA CUCCCUUUGU GUGGGAUGCG UGCAUUUAUC AGAUCAAAAC CAACCCGGUC AGCCUCCCCG CCGGCCGGGG GGGUGGGGCG GCGGCUUUGG UGACUCUAGA UAACCUCGGG CCGAUCGCAG CCCUCCGUGG CGGCGACGAC CCAUUCGAAC GUCUGCCCUA UCAACUUUCG AUGGUAGUCG CCGUGCCUAC CAUGGUGACC ACGGGUGACG GGGAAUCAGG GUUCGAUUCC GGAGAGGGAG CCUGAGAAAC GGCUACCACA UCCAAGGAAG GCAGCAGGCG CGCAAAUUAC CCACUCCCGA CCCGGGGAGG UAGUGACGAA AAAUAACAAU ACAGGACUCU UUCGAGGCCC UGUAAUUGGA AUGAGUCCAC UUUAAAUCCU UUAACGAGGA UCCAUUGGAG GGCAAGUCUG GUCGCAGCAG CCGCGGUAAU UCCAGCUCCA AUAGCGUAUA UUAAAGUUGC UGCAGUUAAA AAGCUCGUAG UUGGAUCUUG UGGAGGGUGC GUAGCGGGCG GUCCGCCGCG AGGCGAGCCA CCGCCCGUCC CCGCCCCUUG CCUCUCGGCG CCCCCUCGAU GCUCUUAGCU GAGUGUCCCG CGGGGCCCGA AGCGUUUACU UUGAAAAAAU UAGAGUGUUC AAAGCAGGCC CGAGCCGCCU GGAUACCGCA GCUAGGAAUA AUGGAAUAGG ACCGCGGUUC UAUUUUGUUG GUUUUCGGAA CUGAGGCCAU GAUUAAGAGG GACGGCCGGG GGCAUUCGUA UUGCGCCGCU AGAGGUGAAA UUCUUGGACC GGCGCAAGAC GGACCAGAGC GAAAGCAUUU GCCAAGAAUG UUUUCAUUAA UCAAGAACGA AAGUCGGAGG UUCGAAGACG AUCAGAUACC GUCGUAGUUC CGACCAUAAA CGAUGCCGAC UGGCGAUGCG GCGGCGUUAU UCCCAUGACC CGCCGGGCAG CUUCCGGGAA ACCAAAGUCU UUGGGUUCCG GGGGGAGUAU GGUUGCAAAG CUGAAACUUA AAGGAAUUGA CGGAAGGGCA CCACCAGGAG UGGAGCCUGC GGCUUAAUUU GAC(C4J)CAACAC GGGAAACCUC ACCCGGCCCG GACACGGACA GGAUUGACAG AUUGAUAGCU CUUUCUCGAU UCUGUGGGUG GUGGUGCAUG GCCGUUCUUA GUUGGUGGAG CGAUUUGUCU GGUUAAUUCC GAUAACGAAC GAGACUCUGG CAUGCUAACU AGUUACGCGA CCCCCGAGCG GUCGGCGUCC CCCAACUUCU UAGAGGGACA AGUGGCGUUC AGCCACCCGA GAUUGAGCAA UAACAGGUCU GUGAUGCCCU UAGAUGUCGG GGCUGCACGC GCGCUACACU GACUGGCUCA GCGUGUGCCU ACCCUACGCC GGCAGGCGCG GGUAACCCGU UGAACCCCAU UCGUGAUGGG GAUCGGGGAU UGCAAUUAUU CCCCAUGAAC GAGGAAUUCC CAGUAAGUGC GGGUCAUAAG CUUGCGUUGA UUAAGUCCCU GCCCUUUGUA CACACCGCCC GUCGCUACUA CCGAUUGGAU GGUUUAGUGA GGCCCUCGGA UCGGCCCGCC GGGGUCGGCC CACGGCCCUG GCGGAGCGCU GAGAAGACGG UCGAACUUGA CUAUCUAGAG GAAGUAAAAG UCGUAACAAG GUUUCCGUAG GUGAACCUGC GGAAGGAUCA UUA
MassTheoretical: 601.015688 kDa
SourceSpecies: Rabbit (rabbit)

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Component #38: protein, Eukaryotic translation initiation factor 2 subunit alpha

ProteinName: Eukaryotic translation initiation factor 2 subunit alpha
Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 32.792867 kDa
SourceSpecies: Rabbit (rabbit)

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Component #39: protein, Eukaryotic translation initiation factor 2 subunit gamma

ProteinName: Eukaryotic translation initiation factor 2 subunit gamma
Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 45.862441 kDa
SourceSpecies: Rabbit (rabbit)

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Component #40: protein, eukaryotic translation initiation factor 1A

ProteinName: eukaryotic translation initiation factor 1A / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 12.788764 kDa
SourceSpecies: Rabbit (rabbit)

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Component #41: protein, ATP-binding cassette sub-family E member 1 (ABCE1)

ProteinName: ATP-binding cassette sub-family E member 1 (ABCE1) / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 66.986812 kDa
SourceSpecies: Rabbit (rabbit)

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Component #42: protein, 40S ribosomal protein uS13

ProteinName: 40S ribosomal protein uS13 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 17.759777 kDa
SourceSpecies: Rabbit (rabbit)

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Component #43: protein, 40S ribosomal protein uS19

ProteinName: 40S ribosomal protein uS19 / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 17.049182 kDa
SourceSpecies: Rabbit (rabbit)

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Component #44: nucleic-acid, beta-globin mRNA

nucleic acidName: beta-globin mRNA / Class: RNA / Structure: OTHER / Synthetic: No
Sequence:
ACCUCAAACA GACACCAUGG UGCACCUGAC UCCUGAGGAG AA
MassTheoretical: 13.475132 kDa
SourceSpecies: Homo sapiens (human)

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Component #45: ligand, IRON/SULFUR CLUSTER

LigandName: IRON/SULFUR CLUSTERIron–sulfur cluster / Number of Copies: 2 / Recombinant expression: No
MassTheoretical: 0.35164 kDa

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Component #46: ligand, MAGNESIUM ION

LigandName: MAGNESIUM ION / Number of Copies: 1 / Recombinant expression: No
MassTheoretical: 2.430505 MDa

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Component #47: ligand, PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER

LigandName: PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER / Number of Copies: 2 / Recombinant expression: No
MassTheoretical: 0.522196 kDa

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Experimental details

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Sample preparation

SpecimenSpecimen state: Particle / Method: cryo EM
Sample solutionpH: 7.4
VitrificationCryogen name: ETHANE / Humidity: 100 %

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
ImagingMicroscope: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 26 e/Å2 / Illumination mode: FLOOD BEAM
LensImaging mode: BRIGHT FIELD
Specimen HolderModel: OTHER
CameraDetector: GATAN K2 SUMMIT (4k x 4k)

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Image processing

ProcessingMethod: single particle reconstruction / Number of projections: 252000
3D reconstructionResolution: 3 Å / Resolution method: FSC 0.143 CUT-OFF

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Atomic model buiding

Modeling #1Refinement protocol: flexible
Output model

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