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- EMDB-11566: Hoxa9 IRES P4 stem-loop bound to the 40S ribosomal subunit - 40S head -

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Basic information

Entry
Database: EMDB / ID: EMD-11566
TitleHoxa9 IRES P4 stem-loop bound to the 40S ribosomal subunit - 40S head
Map data
Sample
  • Complex: Hoxa9 IRES P4 stem-loop bound to the 40S ribosomal subunit
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.1 Å
AuthorsLenarcic T / Boehringer D / Leppek K / Quade N / Fujii K / Susanto TT / Xue S / Genuth NR / Barna M / Ban N
Funding support United States, Switzerland, 3 items
OrganizationGrant numberCountry
National Institutes of Health/National Heart, Lung, and Blood Institute (NIH/NHLBI)R01HD086634 United States
European Molecular Biology Organization (EMBO)ALTF 539-2015
Swiss National Science Foundation310030B_163478 Switzerland
CitationJournal: Mol Cell / Year: 2020
Title: Gene- and Species-Specific Hox mRNA Translation by Ribosome Expansion Segments.
Authors: Kathrin Leppek / Kotaro Fujii / Nick Quade / Teodorus Theo Susanto / Daniel Boehringer / Tea Lenarčič / Shifeng Xue / Naomi R Genuth / Nenad Ban / Maria Barna /
Abstract: Ribosomes have been suggested to directly control gene regulation, but regulatory roles for ribosomal RNA (rRNA) remain largely unexplored. Expansion segments (ESs) consist of multitudes of tentacle- ...Ribosomes have been suggested to directly control gene regulation, but regulatory roles for ribosomal RNA (rRNA) remain largely unexplored. Expansion segments (ESs) consist of multitudes of tentacle-like rRNA structures extending from the core ribosome in eukaryotes. ESs are remarkably variable in sequence and size across eukaryotic evolution with largely unknown functions. In characterizing ribosome binding to a regulatory element within a Homeobox (Hox) 5' UTR, we identify a modular stem-loop within this element that binds to a single ES, ES9S. Engineering chimeric, "humanized" yeast ribosomes for ES9S reveals that an evolutionary change in the sequence of ES9S endows species-specific binding of Hoxa9 mRNA to the ribosome. Genome editing to site-specifically disrupt the Hoxa9-ES9S interaction demonstrates the functional importance for such selective mRNA-rRNA binding in translation control. Together, these studies unravel unexpected gene regulation directly mediated by rRNA and how ribosome evolution drives translation of critical developmental regulators.
History
DepositionJul 31, 2020-
Header (metadata) releaseDec 2, 2020-
Map releaseDec 2, 2020-
UpdateDec 30, 2020-
Current statusDec 30, 2020Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.016
  • Imaged by UCSF Chimera
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  • Surface view colored by height
  • Surface level: 0.016
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_11566.png

Downloads & links

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Map

FileDownload / File: emd_11566.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 1.376 Å
Density
Contour LevelBy AUTHOR: 0.016 / Movie #1: 0.016
Minimum - Maximum-0.059588276 - 0.09677203
Average (Standard dev.)4.4067423e-05 (±0.0026159587)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions256256256
Spacing256256256
CellA=B=C: 352.256 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.3761.3761.376
M x/y/z256256256
origin x/y/z0.0000.0000.000
length x/y/z352.256352.256352.256
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS256256256
D min/max/mean-0.0600.0970.000

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Supplemental data

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Mask #1

Fileemd_11566_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
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Histogram

Histogram (log scale)

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Additional map: #1

Fileemd_11566_additional_1.map
Projections & Slices
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Half map: #2

Fileemd_11566_half_map_1.map
Projections & Slices
AxesZYX

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Histogram

Histogram (log scale)

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Half map: #1

Fileemd_11566_half_map_2.map
Projections & Slices
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Sample components

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Entire : Hoxa9 IRES P4 stem-loop bound to the 40S ribosomal subunit

EntireName: Hoxa9 IRES P4 stem-loop bound to the 40S ribosomal subunit
Components
  • Complex: Hoxa9 IRES P4 stem-loop bound to the 40S ribosomal subunit

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Supramolecule #1: Hoxa9 IRES P4 stem-loop bound to the 40S ribosomal subunit

SupramoleculeName: Hoxa9 IRES P4 stem-loop bound to the 40S ribosomal subunit
type: complex / ID: 1 / Parent: 0
Details: This map contains the 40S ribosomal subunit head. (P4 stem loop RNA was obtained by oligonucleotide synthesis)
Source (natural)Organism: Homo sapiens (human) / Strain: HEK293-6E cells
Molecular weightTheoretical: 1.3 MDa

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 7.6
GridModel: Quantifoil R2/2 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS
VitrificationCryogen name: ETHANE-PROPANE / Chamber humidity: 95 % / Chamber temperature: 278 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average exposure time: 2.5 sec. / Average electron dose: 76.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 1664622
CTF correctionSoftware - Name: Gctf
Startup modelType of model: NONE
Details: 40S structure described in Quade, N., Boehringer, D., Leibundgut, M., van den Heuvel, J., and Ban, N., 2015, Cryo-EM structure of Hepatitis C virus IRES bound to the human ribosome at 3.9 A ...Details: 40S structure described in Quade, N., Boehringer, D., Leibundgut, M., van den Heuvel, J., and Ban, N., 2015, Cryo-EM structure of Hepatitis C virus IRES bound to the human ribosome at 3.9 A resolution. Nat. Commun. 6, 7646:1
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION
Final 3D classificationSoftware - Name: RELION
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION
Final reconstructionResolution.type: BY AUTHOR / Resolution: 4.1 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION / Details: This map contains the 40S ribosomal subunit head. / Number images used: 13530
FSC plot (resolution estimation)

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