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- EMDB-4075: Structure of bacterial 30S-IF1-IF3-mRNA translation pre-initiatio... -

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Basic information

Entry
Database: EMDB / ID: EMD-4075
TitleStructure of bacterial 30S-IF1-IF3-mRNA translation pre-initiation complex (state-1C)
Map dataFor optimal visualization of IF3 N-terminal domain, gauss-filter de map by 1.1 and display it at 0.035 contour
Sample
  • Complex: 30S-IF1-IF3-mRNA pre-initiation complex (state-1C)
    • RNA: x 2 types
    • Protein or peptide: x 22 types
  • Ligand: x 2 types
Function / homology
Function and homology information


translation initiation factor activity / ribosome binding / small ribosomal subunit / tRNA binding / ribosome / rRNA binding / structural constituent of ribosome / translation / ribonucleoprotein complex / mRNA binding ...translation initiation factor activity / ribosome binding / small ribosomal subunit / tRNA binding / ribosome / rRNA binding / structural constituent of ribosome / translation / ribonucleoprotein complex / mRNA binding / zinc ion binding / metal ion binding / cytoplasm
Similarity search - Function
Translation initiation factor IF-1 / Translation initiation factor 3, C-terminal / Translation initiation factor IF-3, C-terminal domain / Translation initiation factor 3 / Translation initiation factor 3, N-terminal / Translation initiation factor 3 (IF-3), N-terminal domain superfamily / Translation initiation factor 3 (IF-3), C-terminal domain superfamily / Translation initiation factor IF-3, N-terminal domain / RNA-binding domain, S1, IF1 type / Translation initiation factor 1A / IF-1 ...Translation initiation factor IF-1 / Translation initiation factor 3, C-terminal / Translation initiation factor IF-3, C-terminal domain / Translation initiation factor 3 / Translation initiation factor 3, N-terminal / Translation initiation factor 3 (IF-3), N-terminal domain superfamily / Translation initiation factor 3 (IF-3), C-terminal domain superfamily / Translation initiation factor IF-3, N-terminal domain / RNA-binding domain, S1, IF1 type / Translation initiation factor 1A / IF-1 / S1 domain IF1 type profile. / 30S ribosomal protein Thx / 30S ribosomal protein Thx / 30S ribosomal protein / Ribosomal protein S14, type Z / Ribosomal protein S14/S29 / Ribosomal protein S3, bacterial-type / Ribosomal protein S6, conserved site / Ribosomal protein S6 signature. / Ribosomal protein S19, bacterial-type / Ribosomal protein S7, bacterial/organellar-type / Ribosomal protein S13, bacterial-type / Ribosomal protein S20 / Ribosomal protein S11, bacterial-type / Ribosomal protein S20 superfamily / Ribosomal protein S20 / Ribosomal protein S9, bacterial/plastid / Ribosomal protein S4, bacterial-type / 30S ribosomal protein S17 / Ribosomal protein S5, bacterial-type / Ribosomal protein S6, plastid/chloroplast / Ribosomal protein S2, bacteria/mitochondria/plastid / Ribosomal protein S18, conserved site / Ribosomal protein S18 signature. / Ribosomal protein S16 / Ribosomal protein S16 / Ribosomal protein S16 domain superfamily / Ribosomal protein S15, bacterial-type / Ribosomal protein S6 / Ribosomal protein S6 / Ribosomal protein S6 superfamily / Ribosomal protein S12, bacterial-type / Translation elongation factor EF1B/ribosomal protein S6 / Ribosomal protein S18 / Ribosomal protein S18 / Ribosomal protein S18 superfamily / K Homology domain / K homology RNA-binding domain / Ribosomal protein S3, conserved site / Ribosomal protein S3 signature. / Ribosomal protein S10, conserved site / Ribosomal protein S10 signature. / Ribosomal protein S14, conserved site / Ribosomal protein S14 signature. / Ribosomal protein S2 signature 1. / KH domain / : / Type-2 KH domain profile. / K Homology domain, type 2 / Ribosomal protein S3, C-terminal / Ribosomal protein S3, C-terminal domain / Ribosomal protein S3, C-terminal domain superfamily / Ribosomal protein S15/S19, conserved site / Ribosomal protein S19 signature. / Ribosomal protein S10 / Ribosomal protein S2, conserved site / Ribosomal protein S19/S15 / Ribosomal protein S19/S15, superfamily / Ribosomal protein S19 / Ribosomal protein S5, N-terminal, conserved site / Ribosomal protein S5 signature. / Ribosomal protein S2 / Ribosomal protein S2, flavodoxin-like domain superfamily / Ribosomal protein S2 / Ribosomal protein S7, conserved site / Ribosomal protein S7 signature. / K homology domain superfamily, prokaryotic type / Ribosomal protein S17, conserved site / Ribosomal protein S17 signature. / Ribosomal protein S13, conserved site / Ribosomal protein S13 signature. / Ribosomal protein S4/S9 N-terminal domain / Ribosomal protein S5 / Ribosomal protein S5, N-terminal / Ribosomal protein S5, N-terminal domain / S5 double stranded RNA-binding domain profile. / Ribosomal protein S13 / 30s ribosomal protein S13, C-terminal / Ribosomal protein S13/S18 / Ribosomal protein S13 family profile. / Ribosomal protein S8 signature. / Ribosomal protein S5, C-terminal / Ribosomal protein S5, C-terminal domain / Ribosomal protein S14 / Ribosomal protein S14p/S29e / Ribosomal protein S4/S9 N-terminal domain / Ribosomal protein S4/S9, N-terminal / Ribosomal protein S4, conserved site / Ribosomal protein S4 signature. / Ribosomal protein S15 signature.
Similarity search - Domain/homology
Small ribosomal subunit protein uS14 / Small ribosomal subunit protein uS17 / Small ribosomal subunit protein uS8 / Small ribosomal subunit protein uS7 / Small ribosomal subunit protein uS2 / Small ribosomal subunit protein uS3 / Small ribosomal subunit protein uS4 / Small ribosomal subunit protein uS9 / Small ribosomal subunit protein uS11 / Small ribosomal subunit protein uS13 ...Small ribosomal subunit protein uS14 / Small ribosomal subunit protein uS17 / Small ribosomal subunit protein uS8 / Small ribosomal subunit protein uS7 / Small ribosomal subunit protein uS2 / Small ribosomal subunit protein uS3 / Small ribosomal subunit protein uS4 / Small ribosomal subunit protein uS9 / Small ribosomal subunit protein uS11 / Small ribosomal subunit protein uS13 / Small ribosomal subunit protein bS20 / Small ribosomal subunit protein uS12 / Small ribosomal subunit protein uS10 / Small ribosomal subunit protein uS19 / 30S ribosomal protein S17 / 30S ribosomal protein S14 type Z / 30S ribosomal protein S8 / Small ribosomal subunit protein uS5 / Translation initiation factor IF-1 / Small ribosomal subunit protein bTHX / Small ribosomal subunit protein uS15 / Small ribosomal subunit protein bS16 / Translation initiation factor IF-3 / Small ribosomal subunit protein bS6 / Small ribosomal subunit protein bS18
Similarity search - Component
Biological speciesThermus thermophilus HB8 (bacteria) / Thermus thermophilus (bacteria) / Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 5.35 Å
AuthorsHussain T / Llacer JL / Wimberly BT / Ramakrishnan V
CitationJournal: Cell / Year: 2016
Title: Large-Scale Movements of IF3 and tRNA during Bacterial Translation Initiation.
Authors: Tanweer Hussain / Jose L Llácer / Brian T Wimberly / Jeffrey S Kieft / V Ramakrishnan /
Abstract: In bacterial translational initiation, three initiation factors (IFs 1-3) enable the selection of initiator tRNA and the start codon in the P site of the 30S ribosomal subunit. Here, we report 11 ...In bacterial translational initiation, three initiation factors (IFs 1-3) enable the selection of initiator tRNA and the start codon in the P site of the 30S ribosomal subunit. Here, we report 11 single-particle cryo-electron microscopy (cryoEM) reconstructions of the complex of bacterial 30S subunit with initiator tRNA, mRNA, and IFs 1-3, representing different steps along the initiation pathway. IF1 provides key anchoring points for IF2 and IF3, thereby enhancing their activities. IF2 positions a domain in an extended conformation appropriate for capturing the formylmethionyl moiety charged on tRNA. IF3 and tRNA undergo large conformational changes to facilitate the accommodation of the formylmethionyl-tRNA (fMet-tRNA(fMet)) into the P site for start codon recognition.
History
DepositionAug 1, 2016-
Header (metadata) releaseAug 17, 2016-
Map releaseOct 5, 2016-
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: 0.05
  • Imaged by UCSF Chimera
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  • Surface view colored by height
  • Surface level: 0.05
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-5lmp
  • Surface level: 0.05
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

FileDownload / File: emd_4075.map.gz / Format: CCP4 / Size: 67 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationFor optimal visualization of IF3 N-terminal domain, gauss-filter de map by 1.1 and display it at 0.035 contour
Voxel sizeX=Y=Z: 1.34 Å
Density
Contour LevelBy AUTHOR: 0.05 / Movie #1: 0.05
Minimum - Maximum-0.08959811 - 0.30097285
Average (Standard dev.)-0.000092469454 (±0.017129645)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions260260260
Spacing260260260
CellA=B=C: 348.4 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.341.341.34
M x/y/z260260260
origin x/y/z0.0000.0000.000
length x/y/z348.400348.400348.400
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS260260260
D min/max/mean-0.0900.301-0.000

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Supplemental data

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Half map: #1

Fileemd_4075_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: #2

Fileemd_4075_half_map_2.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire : 30S-IF1-IF3-mRNA pre-initiation complex (state-1C)

EntireName: 30S-IF1-IF3-mRNA pre-initiation complex (state-1C)
Components
  • Complex: 30S-IF1-IF3-mRNA pre-initiation complex (state-1C)
    • RNA: 16S rRNA
    • Protein or peptide: 30S ribosomal protein S2
    • Protein or peptide: 30S ribosomal protein S3
    • Protein or peptide: 30S ribosomal protein S4
    • Protein or peptide: 30S ribosomal protein S5
    • Protein or peptide: 30S ribosomal protein S6
    • Protein or peptide: 30S ribosomal protein S7
    • Protein or peptide: 30S ribosomal protein S8
    • Protein or peptide: 30S ribosomal protein S9
    • Protein or peptide: 30S ribosomal protein S10
    • Protein or peptide: 30S ribosomal protein S11
    • Protein or peptide: 30S ribosomal protein S12
    • Protein or peptide: 30S ribosomal protein S13
    • Protein or peptide: 30S ribosomal protein S14 type ZRibosome
    • Protein or peptide: 30S ribosomal protein S15
    • Protein or peptide: 30S ribosomal protein S16
    • Protein or peptide: 30S ribosomal protein S17
    • Protein or peptide: 30S ribosomal protein S18
    • Protein or peptide: 30S ribosomal protein S19
    • Protein or peptide: 30S ribosomal protein S20
    • Protein or peptide: 30S ribosomal protein ThxRibosome
    • Protein or peptide: Translation initiation factor IF-1
    • Protein or peptide: Translation initiation factor IF-3
    • RNA: mRNAMessenger RNA
  • Ligand: MAGNESIUM ION
  • Ligand: ZINC ION

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Supramolecule #1: 30S-IF1-IF3-mRNA pre-initiation complex (state-1C)

SupramoleculeName: 30S-IF1-IF3-mRNA pre-initiation complex (state-1C) / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#24
Molecular weightTheoretical: 826.1 KDa

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Macromolecule #1: 16S rRNA

MacromoleculeName: 16S rRNA / type: rna / ID: 1 / Number of copies: 1
Source (natural)Organism: Thermus thermophilus HB8 (bacteria)
Molecular weightTheoretical: 493.958281 KDa
SequenceString: UUUGUUGGAG AGUUUGAUCC UGGCUCAGGG UGAACGCUGG CGGCGUGCCU AAGACAUGCA AGUCGUGCGG GCCGCGGGGU UUUACUCCG UGGUCAGCGG CGGACGGGUG AGUAACGCGU GGGUGACCUA CCCGGAAGAG GGGGACAACC CGGGGAAACU C GGGCUAAU ...String:
UUUGUUGGAG AGUUUGAUCC UGGCUCAGGG UGAACGCUGG CGGCGUGCCU AAGACAUGCA AGUCGUGCGG GCCGCGGGGU UUUACUCCG UGGUCAGCGG CGGACGGGUG AGUAACGCGU GGGUGACCUA CCCGGAAGAG GGGGACAACC CGGGGAAACU C GGGCUAAU CCCCCAUGUG GACCCGCCCC UUGGGGUGUG UCCAAAGGGC UUUGCCCGCU UCCGGAUGGG CCCGCGUCCC AU CAGCUAG UUGGUGGGGU AAUGGCCCAC CAAGGCGACG ACGGGUAGCC GGUCUGAGAG GAUGGCCGGC CACAGGGGCA CUG AGACAC GGGCCCCACU CCUACGGGAG GCAGCAGUUA GGAAUCUUCC GCAAUGGGCG CAAGCCUGAC GGAGCGACGC CGCU UGGAG GAAGAAGCCC UUCGGGGUGU AAACUCCUGA ACCCGGGACG AAACCCCCGA CGAGGGGACU GACGGUACCG GGGUA AUAG CGCCGGCCAA CUCCGUGCCA GCAGCCGCGG UAAUACGGAG GGCGCGAGCG UUACCCGGAU UCACUGGGCG UAAAGG GCG UGUAGGCGGC CUGGGGCGUC CCAUGUGAAA GACCACGGCU CAACCGUGGG GGAGCGUGGG AUACGCUCAG GCUAGAC GG UGGGAGAGGG UGGUGGAAUU CCCGGAGUAG CGGUGAAAUG CGCAGAUACC GGGAGGAACG CCGAUGGCGA AGGCAGCC A CCUGGUCCAC CCGUGACGCU GAGGCGCGAA AGCGUGGGGA GCAAACCGGA UUAGAUACCC GGGUAGUCCA CGCCCUAAA CGAUGCGCGC UAGGUCUCUG GGUCUCCUGG GGGCCGAAGC UAACGCGUUA AGCGCGCCGC CUGGGGAGUA CGGCCGCAAG GCUGAAACU CAAAGGAAUU GACGGGGGCC CGCACAAGCG GUGGAGCAUG UGGUUUAAUU CGAAGCAACG CGAAGAACCU U ACCAGGCC UUGACAUGCU AGGGAACCCG GGUGAAAGCC UGGGGUGCCC CGCGAGGGGA GCCCUAGCAC AGGUGCUGCA UG GCCGUCG UCAGCUCGUG CCGUGAGGUG UUGGGUUAAG UCCCGCAACG AGCGCAACCC CCGCCGUUAG UUGCCAGCGG UUC GGCCGG GCACUCUAAC GGGACUGCCC GCGAAAGCGG GAGGAAGGAG GGGACGACGU CUGGUCAGCA UGGCCCUUAC GGCC UGGGC GACACACGUG CUACAAUGCC CACUACAAAG CGAUGCCACC CGGCAACGGG GAGCUAAUCG CAAAAAGGUG GGCCC AGUU CGGAUUGGGG UCUGCAACCC GACCCCAUGA AGCCGGAAUC GCUAGUAAUC GCGGAUCAGC CAUGCCGCGG UGAAUA CGU UCCCGGGCCU UGUACACACC GCCCGUCACG CCAUGGGAGC GGGCUCUACC CGAAGUCGCC GGGAGCCUAC GGGCAGG CG CCGAGGGUAG GGCCCGUGAC UGGGGCGAAG UCGUAACAAG GUAGCUGUAC CGGAAGGUGC GGCUGGAUCA CCUCCUUU C U

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Macromolecule #24: mRNA

MacromoleculeName: mRNA / type: rna / ID: 24 / Number of copies: 1
Source (natural)Organism: Thermus thermophilus (bacteria)
Molecular weightTheoretical: 12.525466 KDa
SequenceString:
GCUCUUUUAA CAAUUUAUCA GGCAAGGAGG UAAAAAUGU

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Macromolecule #2: 30S ribosomal protein S2

MacromoleculeName: 30S ribosomal protein S2 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 29.317703 KDa
SequenceString: MPVEITVKEL LEAGVHFGHE RKRWNPKFAR YIYAERNGIH IIDLQKTMEE LERTFRFIED LAMRGGTILF VGTKKQAQDI VRMEAERAG MPYVNQRWLG GMLTNFKTIS QRVHRLEELE ALFASPEIEE RPKKEQVRLK HELERLQKYL SGFRLLKRLP D AIFVVDPT ...String:
MPVEITVKEL LEAGVHFGHE RKRWNPKFAR YIYAERNGIH IIDLQKTMEE LERTFRFIED LAMRGGTILF VGTKKQAQDI VRMEAERAG MPYVNQRWLG GMLTNFKTIS QRVHRLEELE ALFASPEIEE RPKKEQVRLK HELERLQKYL SGFRLLKRLP D AIFVVDPT KEAIAVREAR KLFIPVIALA DTDSDPDLVD YIIPGNDDAI RSIQLILSRA VDLIIQARGG VVEPSPSYAL VQ EAEATET PEGESEVEA

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Macromolecule #3: 30S ribosomal protein S3

MacromoleculeName: 30S ribosomal protein S3 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 26.751076 KDa
SequenceString: MGNKIHPIGF RLGITRDWES RWYAGKKQYR HLLLEDQRIR GLLEKELYSA GLARVDIERA ADNVAVTVHV AKPGVVIGRG GERIRVLRE ELAKLTGKNV ALNVQEVQNP NLSAPLVAQR VAEQIERRFA VRRAIKQAVQ RVMESGAKGA KVIVSGRIGG A EQARTEWA ...String:
MGNKIHPIGF RLGITRDWES RWYAGKKQYR HLLLEDQRIR GLLEKELYSA GLARVDIERA ADNVAVTVHV AKPGVVIGRG GERIRVLRE ELAKLTGKNV ALNVQEVQNP NLSAPLVAQR VAEQIERRFA VRRAIKQAVQ RVMESGAKGA KVIVSGRIGG A EQARTEWA AQGRVPLHTL RANIDYGFAL ARTTYGVLGV KAYIFLGEVI GGQKPKARPE LPKAEERPRR RRPAVRVKKE E

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Macromolecule #4: 30S ribosomal protein S4

MacromoleculeName: 30S ribosomal protein S4 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 24.373447 KDa
SequenceString: MGRYIGPVCR LCRREGVKLY LKGERCYSPK CAMERRPYPP GQHGQKRARR PSDYAVRLRE KQKLRRIYGI SERQFRNLFE EASKKKGVT GSVFLGLLES RLDNVVYRLG FAVSRRQARQ LVRHGHITVN GRRVDLPSYR VRPGDEIAVA EKSRNLELIR Q NLEAMKGR ...String:
MGRYIGPVCR LCRREGVKLY LKGERCYSPK CAMERRPYPP GQHGQKRARR PSDYAVRLRE KQKLRRIYGI SERQFRNLFE EASKKKGVT GSVFLGLLES RLDNVVYRLG FAVSRRQARQ LVRHGHITVN GRRVDLPSYR VRPGDEIAVA EKSRNLELIR Q NLEAMKGR KVGPWLSLDV EGMKGKFLRL PDREDLALPV NEQLVIEFYS R

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Macromolecule #5: 30S ribosomal protein S5

MacromoleculeName: 30S ribosomal protein S5 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 17.583416 KDa
SequenceString:
MPETDFEEKM ILIRRTARMQ AGGRRFRFGA LVVVGDRQGR VGLGFGKAPE VPLAVQKAGY YARRNMVEVP LQNGTIPHEI EVEFGASKI VLKPAAPGTG VIAGAVPRAI LELAGVTDIL TKELGSRNPI NIAYATMEAL RQLRTKADVE RLRKGEAHAQ A QG

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Macromolecule #6: 30S ribosomal protein S6

MacromoleculeName: 30S ribosomal protein S6 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 11.988753 KDa
SequenceString:
MRRYEVNIVL NPNLDQSQLA LEKEIIQRAL ENYGARVEKV EELGLRRLAY PIAKDPQGYF LWYQVEMPED RVNDLARELR IRDNVRRVM VVKSQEPFLA NA

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Macromolecule #7: 30S ribosomal protein S7

MacromoleculeName: 30S ribosomal protein S7 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 18.050973 KDa
SequenceString:
MARRRRAEVR QLQPDLVYGD VLVTAFINKI MRDGKKNLAA RIFYDACKII QEKTGQEPLK VFKQAVENVK PRMEVRSRRV GGANYQVPM EVSPRRQQSL ALRWLVQAAN QRPERRAAVR IAHELMDAAE GKGGAVKKKE DVERMAEANR AYAHYRW

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Macromolecule #8: 30S ribosomal protein S8

MacromoleculeName: 30S ribosomal protein S8 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 15.86857 KDa
SequenceString:
MLTDPIADML TRIRNATRVY KESTDVPASR FKEEILRILA REGFIKGYER VDVDGKPYLR VYLKYGPRRQ GPDPRPEQVI HHIRRISKP GRRVYVGVKE IPRVRRGLGI AILSTSKGVL TDREARKLGV GGELICEVW

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Macromolecule #9: 30S ribosomal protein S9

MacromoleculeName: 30S ribosomal protein S9 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 14.410614 KDa
SequenceString:
MEQYYGTGRR KEAVARVFLR PGNGKVTVNG QDFNEYFQGL VRAVAALEPL RAVDALGHFD AYITVRGGGK SGQIDAIKLG IARALVQYN PDYRAKLKPL GFLTRDARVV ERKKYGKHKA RRAPQYSKR

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Macromolecule #10: 30S ribosomal protein S10

MacromoleculeName: 30S ribosomal protein S10 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 11.954968 KDa
SequenceString:
MPKIRIKLRG FDHKTLDASA QKIVEAARRS GAQVSGPIPL PTRVRRFTVI RGPFKHKDSR EHFELRTHNR LVDIINPNRK TIEQLMTLD LPTGVEIEIK TVGGGR

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Macromolecule #11: 30S ribosomal protein S11

MacromoleculeName: 30S ribosomal protein S11 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 13.737868 KDa
SequenceString:
MAKKPSKKKV KRQVASGRAY IHASYNNTIV TITDPDGNPI TWSSGGVIGY KGSRKGTPYA AQLAALDAAK KAMAYGMQSV DVIVRGTGA GREQAIRALQ ASGLQVKSIV DDTPVPHNGC RPKKKFRKAS

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Macromolecule #12: 30S ribosomal protein S12

MacromoleculeName: 30S ribosomal protein S12 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 14.637384 KDa
SequenceString:
MPTINQLVRK GREKVRKKSK VPALKGAPFR RGVCTVVRTV TPKKPNSALR KVAKVRLTSG YEVTAYIPGE GHNLQEHSVV LIRGGRVKD LPGVRYHIVR GVYDAAGVKD RKKSRSKYGT KKPKEAAKTA AKK

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Macromolecule #13: 30S ribosomal protein S13

MacromoleculeName: 30S ribosomal protein S13 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 14.338861 KDa
SequenceString:
MARIAGVEIP RNKRVDVALT YIYGIGKARA KEALEKTGIN PATRVKDLTE AEVVRLREYV ENTWKLEGEL RAEVAANIKR LMDIGCYRG LRHRRGLPVR GQRTRTNART RKGPRKTVAG KKKAPRK

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Macromolecule #14: 30S ribosomal protein S14 type Z

MacromoleculeName: 30S ribosomal protein S14 type Z / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 7.158725 KDa
SequenceString:
MARKALIEKA KRTPKFKVRA YTRCVRCGRA RSVYRFFGLC RICLRELAHK GQLPGVRKAS W

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Macromolecule #15: 30S ribosomal protein S15

MacromoleculeName: 30S ribosomal protein S15 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 10.578407 KDa
SequenceString:
MPITKEEKQK VIQEFARFPG DTGSTEVQVA LLTLRINRLS EHLKVHKKDH HSHRGLLMMV GQRRRLLRYL QREDPERYRA LIEKLGIRG

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Macromolecule #16: 30S ribosomal protein S16

MacromoleculeName: 30S ribosomal protein S16 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 10.409983 KDa
SequenceString:
MVKIRLARFG SKHNPHYRIV VTDARRKRDG KYIEKIGYYD PRKTTPDWLK VDVERARYWL SVGAQPTDTA RRLLRQAGVF RQEAREGA

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Macromolecule #17: 30S ribosomal protein S17

MacromoleculeName: 30S ribosomal protein S17 / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 12.325655 KDa
SequenceString:
MPKKVLTGVV VSDKMQKTVT VLVERQFPHP LYGKVIKRSK KYLAHDPEEK YKLGDVVEII ESRPISKRKR FRVLRLVESG RMDLVEKYL IRRQNYESLS KRGGKA

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Macromolecule #18: 30S ribosomal protein S18

MacromoleculeName: 30S ribosomal protein S18 / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 10.258299 KDa
SequenceString:
MSTKNAKPKK EAQRRPSRKA KVKATLGEFD LRDYRNVEVL KRFLSETGKI LPRRRTGLSA KEQRILAKTI KRARILGLLP FTEKLVRK

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Macromolecule #19: 30S ribosomal protein S19

MacromoleculeName: 30S ribosomal protein S19 / type: protein_or_peptide / ID: 19 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 10.605464 KDa
SequenceString:
MPRSLKKGVF VDDHLLEKVL ELNAKGEKRL IKTWSRRSTI VPEMVGHTIA VYNGKQHVPV YITENMVGHK LGEFAPTRTY RGHGKEAKA TKKK

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Macromolecule #20: 30S ribosomal protein S20

MacromoleculeName: 30S ribosomal protein S20 / type: protein_or_peptide / ID: 20 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 11.736143 KDa
SequenceString:
MAQKKPKRNL SALKRHRQSL KRRLRNKAKK SAIKTLSKKA IQLAQEGKAE EALKIMRKAE SLIDKAAKGS TLHKNAAARR KSRLMRKVR QLLEAAGAPL IGGGLSA

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Macromolecule #21: 30S ribosomal protein Thx

MacromoleculeName: 30S ribosomal protein Thx / type: protein_or_peptide / ID: 21 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 3.35003 KDa
SequenceString:
MGKGDRRTRR GKIWRGTYGK YRPRKKK

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Macromolecule #22: Translation initiation factor IF-1

MacromoleculeName: Translation initiation factor IF-1 / type: protein_or_peptide / ID: 22 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 8.247664 KDa
Recombinant expressionOrganism: Escherichia coli BL21(DE3) (bacteria)
SequenceString:
MAKEKDTIRT EGVVTEALPN ATFRVKLDSG PEILAYISGK MRMHYIRILP GDRVVVEITP YDPTRGRIVY RK

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Macromolecule #23: Translation initiation factor IF-3

MacromoleculeName: Translation initiation factor IF-3 / type: protein_or_peptide / ID: 23 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579
Molecular weightTheoretical: 19.904377 KDa
Recombinant expressionOrganism: Escherichia coli BL21(DE3) (bacteria)
SequenceString:
MKEYLTNERI RAKQVRVVGP DGKQLGIMDT REALRLAQEM DLDLVLVGPN ADPPVARIMD YSKWRYEQQM AEKEARKKAK RTEVKSIKF RVKIDEHDYQ TKLGHIKRFL QEGHKVKVTI MFRGREVAHP ELGERILNRV TEDLKDLAVV EMKPEMLGRD M NMLLAPVK VSA

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Macromolecule #25: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 25 / Number of copies: 108 / Formula: MG
Molecular weightTheoretical: 24.305 Da

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Macromolecule #26: ZINC ION

MacromoleculeName: ZINC ION / type: ligand / ID: 26 / Number of copies: 2 / Formula: ZN
Molecular weightTheoretical: 65.409 Da

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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

Concentration0.08 mg/mL
BufferpH: 7.5
Component:
ConcentrationName
5.0 mMHepes-K
10.0 mMMagnessium acetate
50.0 mMPotassium chloride
10.0 mMAmmonium chloride
6.0 mMBeta mercaptoethanol
GridModel: Quantifoil / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK I

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Electron microscopy

MicroscopeFEI POLARA 300
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 50.0 µm / Calibrated magnification: 104478 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.0 mm / Nominal defocus max: 3.5 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 78000
Sample stageCooling holder cryogen: NITROGEN
TemperatureMin: 90.0 K / Max: 100.0 K
Image recordingFilm or detector model: OTHER / Number grids imaged: 5 / Number real images: 4400 / Average exposure time: 1.1 sec. / Average electron dose: 30.0 e/Å2
Details: Recorded in a FEI Falcon III direct electron detector. Images were collected in movie-mode at 32 frames per second
Experimental equipment
Model: Tecnai Polara / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 666610
CTF correctionSoftware - Name: CTFFIND (ver. 3)
Startup modelType of model: PDB ENTRY
PDB model - PDB ID:
Initial angle assignmentType: PROJECTION MATCHING / Software - Name: RELION (ver. 1.4)
Final 3D classificationSoftware - Name: RELION (ver. 1.4)
Final angle assignmentType: ANGULAR RECONSTITUTION / Software - Name: RELION (ver. 1.4)
Final reconstructionAlgorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 5.35 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 1.4) / Number images used: 18830
DetailsFEI Falcon III

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Atomic model buiding 1

RefinementSpace: RECIPROCAL / Protocol: OTHER / Target criteria: FSC
Output model

PDB-5lmp:
Structure of bacterial 30S-IF1-IF3-mRNA translation pre-initiation complex (state-1C)

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