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- EMDB-10212: Cryo-EM structure of 50S-RsfS complex from Staphylococcus aureus -

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Basic information

Entry
Database: EMDB / ID: EMD-10212
TitleCryo-EM structure of 50S-RsfS complex from Staphylococcus aureus
Map dataCryo-EM map of 50S-RsfS complex from Staphylococcus aureus
Sample
  • Complex: 50S ribosomal subunit complex with stress induced ribosome silencing factor RsfS from Staphylococcus aureus
    • Complex: Ribosomal silencing factor RsfS
      • Protein or peptide: x 1 types
    • Organelle or cellular component: 50S ribosomal subunitProkaryotic large ribosomal subunit
      • RNA: x 2 types
      • Protein or peptide: x 23 types
  • Ligand: x 2 types
Function / homology
Function and homology information


negative regulation of ribosome biogenesis / ribosomal large subunit binding / cytosolic ribosome assembly / large ribosomal subunit rRNA binding / large ribosomal subunit / cytoplasmic translation / 5S rRNA binding / cytosolic large ribosomal subunit / transferase activity / tRNA binding ...negative regulation of ribosome biogenesis / ribosomal large subunit binding / cytosolic ribosome assembly / large ribosomal subunit rRNA binding / large ribosomal subunit / cytoplasmic translation / 5S rRNA binding / cytosolic large ribosomal subunit / transferase activity / tRNA binding / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / ribonucleoprotein complex / translation / mRNA binding / RNA binding / cytoplasm
Similarity search - Function
Protein Iojap/ribosomal silencing factor RsfS / Ribosomal protein L25, long-form / Ribosomal protein L25, beta domain / Ribosomal protein L25, C-terminal / Ribosomal protein TL5, C-terminal domain / Nucleotidyltransferase superfamily / : / Ribosomal protein L16 signature 1. / Ribosomal protein L16, conserved site / Ribosomal protein L16 signature 2. ...Protein Iojap/ribosomal silencing factor RsfS / Ribosomal protein L25, long-form / Ribosomal protein L25, beta domain / Ribosomal protein L25, C-terminal / Ribosomal protein TL5, C-terminal domain / Nucleotidyltransferase superfamily / : / Ribosomal protein L16 signature 1. / Ribosomal protein L16, conserved site / Ribosomal protein L16 signature 2. / Ribosomal protein L17 signature. / Ribosomal L25p family / Ribosomal protein L25 / Ribosomal protein L28/L24 superfamily / Ribosomal protein L25/Gln-tRNA synthetase, N-terminal / Ribosomal protein L32p, bacterial type / Ribosomal protein L25/Gln-tRNA synthetase, anti-codon-binding domain superfamily / Ribosomal protein L28 / Ribosomal protein L35, conserved site / Ribosomal protein L35 signature. / Ribosomal protein L35, non-mitochondrial / Ribosomal protein L18, bacterial-type / Ribosomal protein L19, conserved site / Ribosomal protein L19 signature. / Ribosomal protein L20 signature. / Ribosomal protein L27, conserved site / Ribosomal protein L27 signature. / Ribosomal protein L14P, bacterial-type / Ribosomal protein L34, conserved site / Ribosomal protein L34 signature. / Ribosomal protein L22, bacterial/chloroplast-type / Ribosomal protein L35 / Ribosomal protein L35 superfamily / Ribosomal protein L2, bacterial/organellar-type / Ribosomal protein L35 / Ribosomal L28 family / Ribosomal protein L28/L24 / : / Ribosomal protein L30, bacterial-type / Ribosomal protein L16 / Ribosomal protein L18 / Ribosomal L18 of archaea, bacteria, mitoch. and chloroplast / L28p-like / Ribosomal protein L20 / Ribosomal protein L20 / Ribosomal protein L20, C-terminal / Ribosomal protein L21 / Ribosomal protein L27 / Ribosomal L27 protein / Ribosomal protein L19 / Ribosomal protein L19 superfamily / Ribosomal protein L19 / Ribosomal proteins 50S L24/mitochondrial 39S L24 / Ribosomal protein L17 / Ribosomal protein L17 superfamily / Ribosomal protein L17 / Ribosomal protein L21-like / L21-like superfamily / Ribosomal prokaryotic L21 protein / Ribosomal L32p protein family / Ribosomal protein L24 / Ribosomal protein L32p / Ribosomal protein L34 / Ribosomal protein L34 / Ribosomal protein L13, bacterial-type / Ribosomal protein L3, bacterial/organelle-type / Ribosomal protein L15, bacterial-type / 50S ribosomal protein uL4 / : / Ribosomal protein L15, conserved site / Ribosomal protein L2, conserved site / Ribosomal protein L2 signature. / Ribosomal protein L10e/L16 / Ribosomal protein L10e/L16 superfamily / Ribosomal protein L16p/L10e / Ribosomal protein L2, domain 3 / Ribosomal protein L14P, conserved site / Ribosomal protein L15 signature. / Ribosomal protein L22/L17, conserved site / Ribosomal protein L29/L35 / Ribosomal protein L29/L35 superfamily / Ribosomal L29 protein / Ribosomal protein L13, conserved site / Ribosomal protein L13 signature. / Ribosomal protein L24/L26, conserved site / KOW (Kyprides, Ouzounis, Woese) motif. / Ribosomal Proteins L2, C-terminal domain / Ribosomal protein L2, C-terminal / Ribosomal Proteins L2, C-terminal domain / Ribosomal Proteins L2, RNA binding domain / Ribosomal Proteins L2, RNA binding domain / Ribosomal protein L2 / Ribosomal protein L14 signature. / Ribosomal protein L15 / Ribosomal protein L22 signature. / Ribosomal Proteins L2, RNA binding domain / Ribosomal protein L30, ferredoxin-like fold domain / Ribosomal protein L25/L23 / Ribosomal protein L30, ferredoxin-like fold domain superfamily / Ribosomal protein L14p/L23e
Similarity search - Domain/homology
Large ribosomal subunit protein uL15 / Large ribosomal subunit protein uL30 / Large ribosomal subunit protein uL2 / Large ribosomal subunit protein bL34 / Large ribosomal subunit protein uL3 / Large ribosomal subunit protein uL4 / Large ribosomal subunit protein uL23 / Large ribosomal subunit protein uL22 / Large ribosomal subunit protein uL16 / Large ribosomal subunit protein uL29 ...Large ribosomal subunit protein uL15 / Large ribosomal subunit protein uL30 / Large ribosomal subunit protein uL2 / Large ribosomal subunit protein bL34 / Large ribosomal subunit protein uL3 / Large ribosomal subunit protein uL4 / Large ribosomal subunit protein uL23 / Large ribosomal subunit protein uL22 / Large ribosomal subunit protein uL16 / Large ribosomal subunit protein uL29 / Large ribosomal subunit protein uL14 / Large ribosomal subunit protein uL24 / Large ribosomal subunit protein uL18 / Large ribosomal subunit protein bL17 / Large ribosomal subunit protein uL13 / Large ribosomal subunit protein bL35 / Large ribosomal subunit protein bL20 / Large ribosomal subunit protein bL21 / Large ribosomal subunit protein bL27 / Large ribosomal subunit protein bL19 / Large ribosomal subunit protein bL28 / Large ribosomal subunit protein bL32 / Large ribosomal subunit protein bL25 / Ribosomal silencing factor RsfS
Similarity search - Component
Biological speciesStaphylococcus aureus subsp. aureus NCTC 8325 (bacteria) / Staphylococcus aureus (strain NCTC 8325) (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.23 Å
AuthorsKhusainov I / Pellegrino S / Yusupova G / Yusupov M / Fatkhullin B
Funding support France, 3 items
OrganizationGrant numberCountry
French National Research AgencyANR-15-CE11-0021-01 France
French National Research AgencyLabex: ANR-10-LABX- 0036_NETRNA France
French National Research AgencyANR-16-CE11-0007-01 France
CitationJournal: Nat Commun / Year: 2020
Title: Mechanism of ribosome shutdown by RsfS in Staphylococcus aureus revealed by integrative structural biology approach.
Authors: Iskander Khusainov / Bulat Fatkhullin / Simone Pellegrino / Aydar Bikmullin / Wen-Ti Liu / Azat Gabdulkhakov / Amr Al Shebel / Alexander Golubev / Denis Zeyer / Natalie Trachtmann / Georg A ...Authors: Iskander Khusainov / Bulat Fatkhullin / Simone Pellegrino / Aydar Bikmullin / Wen-Ti Liu / Azat Gabdulkhakov / Amr Al Shebel / Alexander Golubev / Denis Zeyer / Natalie Trachtmann / Georg A Sprenger / Shamil Validov / Konstantin Usachev / Gulnara Yusupova / Marat Yusupov /
Abstract: For the sake of energy preservation, bacteria, upon transition to stationary phase, tone down their protein synthesis. This process is favored by the reversible binding of small stress-induced ...For the sake of energy preservation, bacteria, upon transition to stationary phase, tone down their protein synthesis. This process is favored by the reversible binding of small stress-induced proteins to the ribosome to prevent unnecessary translation. One example is the conserved bacterial ribosome silencing factor (RsfS) that binds to uL14 protein onto the large ribosomal subunit and prevents its association with the small subunit. Here we describe the binding mode of Staphylococcus aureus RsfS to the large ribosomal subunit and present a 3.2 Å resolution cryo-EM reconstruction of the 50S-RsfS complex together with the crystal structure of uL14-RsfS complex solved at 2.3 Å resolution. The understanding of the detailed landscape of RsfS-uL14 interactions within the ribosome shed light on the mechanism of ribosome shutdown in the human pathogen S. aureus and might deliver a novel target for pharmacological drug development and treatment of bacterial infections.
History
DepositionAug 12, 2019-
Header (metadata) releaseApr 8, 2020-
Map releaseApr 8, 2020-
UpdateJul 8, 2020-
Current statusJul 8, 2020Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.027
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by height
  • Surface level: 0.027
  • Imaged by UCSF Chimera
  • Download
  • Surface view with fitted model
  • Atomic models: PDB-6sj6
  • Surface level: 0.027
  • Imaged by UCSF Chimera
  • Download
Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_10212.png

Downloads & links

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Map

FileDownload / File: emd_10212.map.gz / Format: CCP4 / Size: 163.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationCryo-EM map of 50S-RsfS complex from Staphylococcus aureus
Voxel sizeX=Y=Z: 0.858 Å
Density
Contour LevelBy AUTHOR: 0.027 / Movie #1: 0.027
Minimum - Maximum-0.10070784 - 0.16990301
Average (Standard dev.)0.0009123183 (±0.006633047)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions350350350
Spacing350350350
CellA=B=C: 300.3 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z0.8580.8580.858
M x/y/z350350350
origin x/y/z0.0000.0000.000
length x/y/z300.300300.300300.300
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS350350350
D min/max/mean-0.1010.1700.001

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Supplemental data

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Sample components

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Entire : 50S ribosomal subunit complex with stress induced ribosome silenc...

EntireName: 50S ribosomal subunit complex with stress induced ribosome silencing factor RsfS from Staphylococcus aureus
Components
  • Complex: 50S ribosomal subunit complex with stress induced ribosome silencing factor RsfS from Staphylococcus aureus
    • Complex: Ribosomal silencing factor RsfS
      • Protein or peptide: Ribosomal silencing factor RsfS
    • Organelle or cellular component: 50S ribosomal subunitProkaryotic large ribosomal subunit
      • RNA: 23S ribosomal RNA
      • RNA: 5S ribosomal RNA
      • Protein or peptide: 50S ribosomal protein L2
      • Protein or peptide: 50S ribosomal protein L3
      • Protein or peptide: 50S ribosomal protein L4
      • Protein or peptide: 50S ribosomal protein L13
      • Protein or peptide: 50S ribosomal protein L14
      • Protein or peptide: 50S ribosomal protein L15
      • Protein or peptide: 50S ribosomal protein L16
      • Protein or peptide: 50S ribosomal protein L17
      • Protein or peptide: 50S ribosomal protein L18
      • Protein or peptide: 50S ribosomal protein L19
      • Protein or peptide: 50S ribosomal protein L20
      • Protein or peptide: 50S ribosomal protein L21
      • Protein or peptide: 50S ribosomal protein L22
      • Protein or peptide: 50S ribosomal protein L23
      • Protein or peptide: 50S ribosomal protein L24
      • Protein or peptide: 50S ribosomal protein L25
      • Protein or peptide: 50S ribosomal protein L27
      • Protein or peptide: 50S ribosomal protein L28
      • Protein or peptide: 50S ribosomal protein L29
      • Protein or peptide: 50S ribosomal protein L30
      • Protein or peptide: 50S ribosomal protein L32
      • Protein or peptide: 50S ribosomal protein L34
      • Protein or peptide: 50S ribosomal protein L35
  • Ligand: MAGNESIUM ION
  • Ligand: POTASSIUM IONPotassium

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Supramolecule #1: 50S ribosomal subunit complex with stress induced ribosome silenc...

SupramoleculeName: 50S ribosomal subunit complex with stress induced ribosome silencing factor RsfS from Staphylococcus aureus
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#26
Molecular weightExperimental: 1.35 MDa

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Supramolecule #3: Ribosomal silencing factor RsfS

SupramoleculeName: Ribosomal silencing factor RsfS / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #26
Source (natural)Organism: Staphylococcus aureus subsp. aureus NCTC 8325 (bacteria)
Recombinant expressionOrganism: Escherichia coli BL21(DE3) (bacteria)

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Supramolecule #2: 50S ribosomal subunit

SupramoleculeName: 50S ribosomal subunit / type: organelle_or_cellular_component / ID: 2 / Parent: 1 / Macromolecule list: #1-#25
Source (natural)Organism: Staphylococcus aureus subsp. aureus NCTC 8325 (bacteria)

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Macromolecule #1: 23S ribosomal RNA

MacromoleculeName: 23S ribosomal RNA / type: rna / ID: 1 / Number of copies: 1
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 946.696625 KDa
SequenceString: GAUUAAGUUA UUAAGGGCGC ACGGUGGAUG CCUUGGCACU AGAAGCCGAU GAAGGACGUU ACUAACGACG AUAUGCUUUG GGGAGCUGU AAGUAAGCUU UGAUCCAGAG AUUUCCGAAU GGGGAAACCC AGCAUGAGUU AUGUCAUGUU AUCGAUAUGU G AAUACAUA ...String:
GAUUAAGUUA UUAAGGGCGC ACGGUGGAUG CCUUGGCACU AGAAGCCGAU GAAGGACGUU ACUAACGACG AUAUGCUUUG GGGAGCUGU AAGUAAGCUU UGAUCCAGAG AUUUCCGAAU GGGGAAACCC AGCAUGAGUU AUGUCAUGUU AUCGAUAUGU G AAUACAUA GCAUAUCAGA AGGCACACCC GGAGAACUGA AACAUCUUAG UACCCGGAGG AAGAGAAAGA AAAUUCGAUU CC CUUAGUA GCGGCGAGCG AAACGGGAAG AGCCCAAACC AACAAGCUUG CUUGUUGGGG UUGUAGGACA CUCUAUACGG AGU UACAAA GGACGACAUU AGACGAAUCA UCUGGAAAGA UGAAUCAAAG AAGGUAAUAA UCCUGUAGUC GAAAAUGUUG UCUC UCUUG AGUGGAUCCU GAGUACGACG GAGCACGUGA AAUUCCGUCG GAAUCUGGGA GGACCAUCUC CUAAGGCUAA AUACU CUCU AGUGACCGAU AGUGAACCAG UACCGUGAGG GAAAGGUGAA AAGCACCCCG GAAGGGGAGU GAAAUAGAAC CUGAAA CCG UGUGCUUACA AGUAGUCAGA GCCCGUUAAU GGGUGAUGGC GUGCCUUUUG UAGAAUGAAC CGGCGAGUUA CGAUUUG AU GCAAGGUUAA GCAGUAAAUG UGGAGCCGUA GCGAAAGCGA GUCUGAAUAG GGCGUUUAGU AUUUGGUCGU AGACCCGA A ACCAGGUGAU CUACCCUUGG UCAGGUUGAA GUUCAGGUAA CACUGAAUGG AGGACCGAAC CGACUUACGU UGAAAAGUG AGCGGAUGAA CUGAGGGUAG CGGAGAAAUU CCAAUCGAAC CUGGAGAUAG CUGGUUCUCU CCGAAAUAGC UUUAGGGCUA GCCUCAAGU GAUGAUUAUU GGAGGUAGAG CACUGUUUGG ACGAGGGGCC CCUCUCGGGU UACCGAAUUC AGACAAACUC C GAAUGCCA AUUAAUUUAA CUUGGGAGUC AGAACAUGGG UGAUAAGGUC CGUGUUCGAA AGGGAAACAG CCCAGACCAC CA GCUAAGG UCCCAAAAUA UAUGUUAAGU GGAAAAGGAU GUGGCGUUGC CCAGACAACU AGGAUGUUGG CUUAGAAGCA GCC AUCAUU UAAAGAGUGC GUAAUAGCUC ACUAGUCGAG UGACACUGCG CCGAAAAUGU ACCGGGGCUA AACAUAUUAC CGAA GCUGU GGAUUGUCCU UUGGACAAUG GUAGGAGAGC GUUCUAAGGG CGUUGAAGCA UGAUCGUAAG GACAUGUGGA GCGCU UAGA AGUGAGAAUG CCGGUGUGAG UAGCGAAAGA CGGGUGAGAA UCCCGUCCAC CGAUUGACUA AGGUUUCCAG AGGAAG GCU CGUCCGCUCU GGGUUAGUCG GGUCCUAAGC UGAGGCCGAC AGGCGUAGGC GAUGGAUAAC AGGUUGAUAU UCCUGUA CC ACCUAUAAUC GUUUUAAUCG AUGGGGGGAC GCAGUAGGAU AGGCGAAGCG UGCGAUUGGA UUGCACGUCU AAGCAGUA A GGCUGAGUAU UAGGCAAAUC CGGUACUCGU UAAGGCUGAG CUGUGAUGGG GAGAAGACAU UGUGUCUUCG AGUCGUUGA UUUCACACUG CCGAGAAAAG CCUCUAGAUA GAAAAUAGGU GCCCGUACCG CAAACCGACA CAGGUAGUCA AGAUGAGAAU UCUAAGGUG AGCGAGCGAA CUCUCGUUAA GGAACUCGGC AAAAUGACCC CGUAACUUCG GGAGAAGGGG UGCUCUUUAG G GUUAACGC CCAGAAGAGC CGCAGUGAAU AGGCCCAAGC GACUGUUUAU CAAAAACACA GGUCUCUGCU AAACCGUAAG GU GAUGUAU AGGGGCUGAC GCCUGCCCGG UGCUGGAAGG UUAAGAGGAG UGGUUAGCUU CUGCGAAGCU ACGAAUCGAA GCC CCAGUA AACGGCGGCC GUAACUAUAA CGGUCCUAAG GUAGCGAAAU UCCUUGUCGG GUAAGUUCCG ACCCGCACGA AAGG CGUAA CGAUUUGGGC ACUGUCUCAA CGAGAGACUC GGUGAAAUCA UAGUACCUGU GAAGAUGCAG GUUACCCGCG ACAGG ACGG AAAGACCCCG UGGAGCUUUA CUGUAGCCUG AUAUUGAAAU UCGGCACAGC UUGUACAGGA UAGGUAGGAG CCUUUG AAA CGUGAGCGCU AGCUUACGUG GAGGCGCUGG UGGGAUACUA CCCUAGCUGU GUUGGCUUUC UAACCCGCAC CACUUAU CG UGGUGGGAGA CAGUGUCAGG CGGGCAGUUU GACUGGGGCG GUCGCCUCCU AAAAGGUAAC GGAGGCGCUC AAAGGUUC C CUCAGAAUGG UUGGAAAUCA UUCAUAGAGU GUAAAGGCAU AAGGGAGCUU GACUGCGAGA CCUACAAGUC GAGCAGGGU CGAAAGACGG ACUUAGUGAU CCGGUGGUUC CGCAUGGAAG GGCCAUCGCU CAACGGAUAA AAGCUACCCC GGGGAUAACA GGCUUAUCU CCCCCAAGAG UUCACAUCGA CGGGGAGGUU UGGCACCUCG AUGUCGGCUC AUCGCAUCCU GGGGCUGUAG U CGGUCCCA AGGGUUGGGC UGUUCGCCCA UUAAAGCGGU ACGCGAGCUG GGUUCAGAAC GUCGUGAGAC AGUUCGGUCC CU AUCCGUC GUGGGCGUAG GAAAUUUGAG AGGAGCUGUC CUUAGUACGA GAGGACCGGG AUGGACAUAC CUCUGGUGUA CCA GUUGUC GUGCCAACGG CAUAGCUGGG UAGCUAUGUG UGGACGGGAU AAGUGCUGAA AGCAUCUAAG CAUGAAGCCC CCCU CAAGA UGAGAUUUCC CAACUUCGGU UAUAAGAUCC CUCAAAGAUG AUGAGGUUAA UAGGUUCGAG GUGGAAGCAU GGUGA CAUG UGGAGCUGAC GAAUACUAAU CGAUCGAAGA CUUAAUCAA

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Macromolecule #2: 5S ribosomal RNA

MacromoleculeName: 5S ribosomal RNA / type: rna / ID: 2 / Number of copies: 1
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 36.974945 KDa
SequenceString:
UCUGGUGACU AUAGCAAGGA GGUCACACCU GUUCCCAUGC CGAACACAGA AGUUAAGCUC CUUAGCGUCG AUGGUAGUCG AACUUACGU UCCGCUAGAG UAGAACGUUG CCAGGC

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Macromolecule #3: 50S ribosomal protein L2

MacromoleculeName: 50S ribosomal protein L2 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 30.217164 KDa
SequenceString: MAIKKYKPIT NGRRNMTSLD FAEITKTTPE KSLLKPLPKK AGRNNQGKLT VRHHGGGHKR QYRVIDFKRN KDGINAKVDS IQYDPNRSA NIALVVYADG EKRYIIAPKG LEVGQIVESG AEADIKVGNA LPLQNIPVGT VVHNIELKPG KGGQIARSAG A SAQVLGKE ...String:
MAIKKYKPIT NGRRNMTSLD FAEITKTTPE KSLLKPLPKK AGRNNQGKLT VRHHGGGHKR QYRVIDFKRN KDGINAKVDS IQYDPNRSA NIALVVYADG EKRYIIAPKG LEVGQIVESG AEADIKVGNA LPLQNIPVGT VVHNIELKPG KGGQIARSAG A SAQVLGKE GKYVLIRLRS GEVRMILSTC RATIGQVGNL QHELVNVGKA GRSRWKGIRP TVRGSVMNPN DHPHGGGEGR AP IGRPSPM SPWGKPTLGK KTRRGKKSSD KLIVRGRKKK

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Macromolecule #4: 50S ribosomal protein L3

MacromoleculeName: 50S ribosomal protein L3 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 23.760256 KDa
SequenceString: MTKGILGRKI GMTQVFGENG ELIPVTVVEA KENVVLQKKT VEVDGYNAIQ VGFEDKKAYK KDAKSNKYAN KPAEGHAKKA DAAPKRFIR EFRNVDVDAY EVGQEVSVDT FVAGDVIDVT GVSKGKGFQG AIKRHGQSRG PMSHGSHFHR APGSVGMASD A SRVFKGQK ...String:
MTKGILGRKI GMTQVFGENG ELIPVTVVEA KENVVLQKKT VEVDGYNAIQ VGFEDKKAYK KDAKSNKYAN KPAEGHAKKA DAAPKRFIR EFRNVDVDAY EVGQEVSVDT FVAGDVIDVT GVSKGKGFQG AIKRHGQSRG PMSHGSHFHR APGSVGMASD A SRVFKGQK MPGRMGGNTV TVQNLEVVQV DTENKVILVK GNVPGPKKGL VEIRTSIKKG NK

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Macromolecule #5: 50S ribosomal protein L4

MacromoleculeName: 50S ribosomal protein L4 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 22.495697 KDa
SequenceString: MANYDVLKLD GTKSGSIELS DAVFGIEPNN SVLFEAINLQ RASLRQGTHA VKNRSAVSGG GRKPWKQKGT GRARQGTIRA PQWRGGGIV FGPTPRSYAY KMPKKMRRLA LRSALSFKAQ ENGLTVVDAF NFEAPKTKEF KNVLSTLEQP KKVLVVTENE D VNVELSAR ...String:
MANYDVLKLD GTKSGSIELS DAVFGIEPNN SVLFEAINLQ RASLRQGTHA VKNRSAVSGG GRKPWKQKGT GRARQGTIRA PQWRGGGIV FGPTPRSYAY KMPKKMRRLA LRSALSFKAQ ENGLTVVDAF NFEAPKTKEF KNVLSTLEQP KKVLVVTENE D VNVELSAR NIPGVQVTTA QGLNVLDITN ADSLVITEAA AKKVEEVLG

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Macromolecule #6: 50S ribosomal protein L13

MacromoleculeName: 50S ribosomal protein L13 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 16.359427 KDa
SequenceString:
MRQTFMANES NIERKWYVID AEGQTLGRLS SEVASILRGK NKVTYTPHVD TGDYVIVINA SKIEFTGNKE TDKVYYRHSN HPGGIKSIT AGELRRTNPE RLIENSIKGM LPSTRLGEKQ GKKLFVYGGA EHPHAAQQPE NYELRG

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Macromolecule #7: 50S ribosomal protein L14

MacromoleculeName: 50S ribosomal protein L14 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 13.157342 KDa
SequenceString:
MIQQETRLKV ADNSGAREVL TIKVLGGSGR KTANIGDVIV CTVKNATPGG VVKKGDVVKA VIVRTKSGVR RNDGSYIKFD ENACVIIRD DKGPRGTRIF GPVARELREG NFMKIVSLAP EVL

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Macromolecule #8: 50S ribosomal protein L15

MacromoleculeName: 50S ribosomal protein L15 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 15.62889 KDa
SequenceString:
MKLHELKPAE GSRKERNRVG RGVATGNGKT SGRGHKGQKA RSGGGVRPGF EGGQLPLFRR LPKRGFTNIN RKEYAIVNLD QLNKFEDGT EVTPALLVES GVVKNEKSGI KILGNGSLDK KLTVKAHKFS ASAAEAIDAK GGAHEVI

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Macromolecule #9: 50S ribosomal protein L16

MacromoleculeName: 50S ribosomal protein L16 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 16.274049 KDa
SequenceString:
MLLPKRVKYR RQHRPKTTGR SKGGNYVTFG EFGLQATTTS WITSRQIESA RIAMTRYMKR GGKVWIKIFP HTPYTKKPLE VRMGAGKGA VEGWIAVVKP GRILFEVAGV SEEVAREALR LASHKLPVKT KFVKREELGG ETNES

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Macromolecule #10: 50S ribosomal protein L17

MacromoleculeName: 50S ribosomal protein L17 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 13.771773 KDa
SequenceString:
MGYRKLGRTS DQRKAMLRDL ATSLIISERI ETTEARAKEV RSVVEKLITL GKKGDLASRR NAAKTLRNVE ILNEDETTQT ALQKLFGEI AERYTERQGG YTRILKQGPR RGDGAESVII ELV

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Macromolecule #11: 50S ribosomal protein L18

MacromoleculeName: 50S ribosomal protein L18 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 13.124093 KDa
SequenceString:
MISKIDKNKV RLKRHARVRT NLSGTAEKPR LNVYRSNKHI YAQIIDDNKG VTLAQASSKD SDIATTATKV ELATKVGEAI AKKAADKGI KEIVFDRGGY LYHGRVKALA EAARESGLEF

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Macromolecule #12: 50S ribosomal protein L19

MacromoleculeName: 50S ribosomal protein L19 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 13.392771 KDa
SequenceString:
MTNHKLIEAV TKSQLRTDLP SFRPGDTLRV HVRIIEGTRE RIQVFEGVVI KRRGGGVSET FTVRKISSGV GVERTFPLHT PKIEKIEVK RRGKVRRAKL YYLRSLRGKA ARIQEIR

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Macromolecule #13: 50S ribosomal protein L20

MacromoleculeName: 50S ribosomal protein L20 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 13.720295 KDa
SequenceString:
MPRVKGGTVT RARRKKTIKL AKGYFGSKHT LYKVAKQQVM KSGQYAFRDR RQRKRDFRKL WITRINAAAR QHEMSYSRLM NGLKKAGID INRKMLSEIA ISDEKAFAQL VTKAKDALK

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Macromolecule #14: 50S ribosomal protein L21

MacromoleculeName: 50S ribosomal protein L21 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 11.354081 KDa
SequenceString:
MFAIIETGGK QIKVEEGQEI FVEKLDVNEG DTFTFDKVLF VGGDSVKVGA PTVEGATVTA TVNKQGRGKK ITVFTYKRRK NSKRKKGHR QPYTKLTIDK INA

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Macromolecule #15: 50S ribosomal protein L22

MacromoleculeName: 50S ribosomal protein L22 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 12.857922 KDa
SequenceString:
MEAKAVARTI RIAPRKVRLV LDLIRGKNAA EAIAILKLTN KASSPVIEKV LMSALANAEH NYDMNTDELV VKEAYANEGP TLKRFRPRA QGRASAINKR TSHITIVVSD GKEEAKEA

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Macromolecule #16: 50S ribosomal protein L23

MacromoleculeName: 50S ribosomal protein L23 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 10.625398 KDa
SequenceString:
MEARDILKRP VITEKSSEAM AEDKYTFDVD TRVNKTQVKM AVEEIFNVKV ASVNIMNYKP KKKRMGRYQG YTNKRRKAIV TLKEGSIDL FN

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Macromolecule #17: 50S ribosomal protein L24

MacromoleculeName: 50S ribosomal protein L24 / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 11.561504 KDa
SequenceString:
MHIKKGDNVK VIAGKDKGKE GKVIATLPKK DRVVVEGVNI MKKHQKPTQL NPEGGILETE AAIHVSNVQL LDPKTNEPTR VGYKFVDGK KVRIAKKSGE EIKSNN

+
Macromolecule #18: 50S ribosomal protein L25

MacromoleculeName: 50S ribosomal protein L25 / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 23.810609 KDa
SequenceString: MASLKSIIRQ GKQTRSDLKQ LRKSGKVPAV VYGYGTKNVS VKVDEVEFIK VIREVGRNGV IELGVGSKTI KVMVADYQFD PLKNQITHI DFLAINMSEE RTVEVPVQLV GEAVGAKEGG VVEQPLFNLE VTATPDNIPE AIEVDITELN INDSLTVADV K VTGDFKIE ...String:
MASLKSIIRQ GKQTRSDLKQ LRKSGKVPAV VYGYGTKNVS VKVDEVEFIK VIREVGRNGV IELGVGSKTI KVMVADYQFD PLKNQITHI DFLAINMSEE RTVEVPVQLV GEAVGAKEGG VVEQPLFNLE VTATPDNIPE AIEVDITELN INDSLTVADV K VTGDFKIE NDSAESVVTV VAPTEEPTEE EIEAMEGEQQ TEEPEVVGES KEDEEKTEE

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Macromolecule #19: 50S ribosomal protein L27

MacromoleculeName: 50S ribosomal protein L27 / type: protein_or_peptide / ID: 19 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 10.334798 KDa
SequenceString:
MLKLNLQFFA SKKGVSSTKN GRDSESKRLG AKRADGQFVT GGSILYRQRG TKIYPGENVG RGGDDTLFAK IDGVVKFERK GRDKKQVSV YAVAE

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Macromolecule #20: 50S ribosomal protein L28

MacromoleculeName: 50S ribosomal protein L28 / type: protein_or_peptide / ID: 20 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 6.995289 KDa
SequenceString:
MGKQCFVTGR KASTGNRRSH ALNSTKRRWN ANLQKVRILV DGKPKKVWVS ARALKSGKVT RV

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Macromolecule #21: 50S ribosomal protein L29

MacromoleculeName: 50S ribosomal protein L29 / type: protein_or_peptide / ID: 21 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 8.105266 KDa
SequenceString:
MKAKEIRDLT TSEIEEQIKS SKEELFNLRF QLATGQLEET ARIRTVRKTI ARLKTVARER EIEQSKANQ

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Macromolecule #22: 50S ribosomal protein L30

MacromoleculeName: 50S ribosomal protein L30 / type: protein_or_peptide / ID: 22 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 6.565683 KDa
SequenceString:
MAKLQITLTR SVIGRPETQR KTVEALGLKK TNSSVVVEDN PAIRGQINKV KHLVTVEEK

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Macromolecule #23: 50S ribosomal protein L32

MacromoleculeName: 50S ribosomal protein L32 / type: protein_or_peptide / ID: 23 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 6.657803 KDa
SequenceString:
MAVPKRRTSK TRKNKRRTHF KISVPGMTEC PNCGREYKLS HRVCKNCGSY NGEEVAAK

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Macromolecule #24: 50S ribosomal protein L34

MacromoleculeName: 50S ribosomal protein L34 / type: protein_or_peptide / ID: 24 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 5.454642 KDa
SequenceString:
MVKRTYQPNK RKHSKVHGFR KRMSTKNGRK VLARRRRKGR KVLSA

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Macromolecule #25: 50S ribosomal protein L35

MacromoleculeName: 50S ribosomal protein L35 / type: protein_or_peptide / ID: 25 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Molecular weightTheoretical: 7.722368 KDa
SequenceString:
MPKMKTHRGA AKRVKRTASG QLKRSRAFTS HLFANKSTKQ KRQLRKARLV SKSDMKRVKQ LLAYKK

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Macromolecule #26: Ribosomal silencing factor RsfS

MacromoleculeName: Ribosomal silencing factor RsfS / type: protein_or_peptide / ID: 26 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Staphylococcus aureus (strain NCTC 8325) (bacteria)
Strain: NCTC 8325
Molecular weightTheoretical: 13.497068 KDa
Recombinant expressionOrganism: Escherichia coli BL21(DE3) (bacteria)
SequenceString:
MNSQELLAIA VDAIDNKKGE DTISLEMKGI SDMTDYFVVT HGNNERQVQA IARAVKEVAN EQNIEVKRME GYNEARWILI DLADVVVHV FHKDERNYYN IEKLYQDAPL ESYSQVAY

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Macromolecule #27: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 27 / Number of copies: 56 / Formula: MG
Molecular weightTheoretical: 24.305 Da

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Macromolecule #28: POTASSIUM ION

MacromoleculeName: POTASSIUM ION / type: ligand / ID: 28 / Number of copies: 3 / Formula: K
Molecular weightTheoretical: 39.098 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

Concentration0.7 mg/mL
BufferpH: 7.5
Component:
ConcentrationFormulaName
10.0 mMC8H18N2O4SHepes
50.0 mMKClpotassium chloride
10.0 mMNH4Clammonium chloride
10.0 mMMg(CH3OO)2magnesium acetate
GridModel: Quantifoil R2/2 / Material: COPPER / Mesh: 400 / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 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 / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.2 µm / Nominal magnification: 96000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: FEI FALCON III (4k x 4k) / Number grids imaged: 2 / Number real images: 1856 / Average exposure time: 43.0 sec. / Average electron dose: 24.6 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 172729
CTF correctionSoftware - Name: Gctf
Startup modelType of model: EMDB MAP
EMDB ID:

6177

Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 2.1)
Final 3D classificationSoftware - Name: RELION (ver. 2.1)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 2.1)
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.23 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 2.1) / Number images used: 83885
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelPDB ID:

5li0

RefinementSpace: REAL / Protocol: FLEXIBLE FIT
Output model

PDB-6sj6:
Cryo-EM structure of 50S-RsfS complex from Staphylococcus aureus

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