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- EMDB-37564: Cryo- EM structure of Mycobacterium smegmatis 30S ribosomal subun... -

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Basic information

Entry
Database: EMDB / ID: EMD-37564
TitleCryo- EM structure of Mycobacterium smegmatis 30S ribosomal subunit (body 2) of 70S ribosome, E- tRNA and RafH.
Map dataMycobacterium smegmatis 30S ribosome (body 2) of 70S ribosome, E t-RNA and RafH.
Sample
  • Complex: 70S Ribosome + RafH protein
    • Complex: 70S RibosomeRibosome
      • RNA: x 2 types
      • Protein or peptide: x 20 types
    • Complex: RafH
      • Protein or peptide: x 1 types
KeywordsRibosome / protein synthesis / Mycobacterium smegmatis / hibernation promotion factor / HPF / RafH / hypoxia stress / Cryo- EM / Single particle reconstruction
Function / homology
Function and homology information


primary metabolic process / regulation of translation / small ribosomal subunit / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / ribonucleoprotein complex / translation / mRNA binding ...primary metabolic process / regulation of translation / small ribosomal subunit / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / ribonucleoprotein complex / translation / mRNA binding / metal ion binding / cytoplasm
Similarity search - Function
Sigma 54 modulation/S30EA ribosomal protein, C-terminal / Sigma 54 modulation/S30EA ribosomal protein, C-terminal domain superfamily / Sigma 54 modulation/S30EA ribosomal protein C terminus / Ribosome hibernation promoting factor/RaiA / Ribosome hibernation promotion factor-like / Sigma 54 modulation protein / S30EA ribosomal protein / Mitochondrial mRNA-processing protein COX24, C-terminal / Mitochondrial mRNA-processing protein COX24, C-terminal / Mitochondrial domain of unknown function (DUF1713) / Ribosomal protein S14, bacterial/plastid ...Sigma 54 modulation/S30EA ribosomal protein, C-terminal / Sigma 54 modulation/S30EA ribosomal protein, C-terminal domain superfamily / Sigma 54 modulation/S30EA ribosomal protein C terminus / Ribosome hibernation promoting factor/RaiA / Ribosome hibernation promotion factor-like / Sigma 54 modulation protein / S30EA ribosomal protein / Mitochondrial mRNA-processing protein COX24, C-terminal / Mitochondrial mRNA-processing protein COX24, C-terminal / Mitochondrial domain of unknown function (DUF1713) / Ribosomal protein S14, bacterial/plastid / Ribosomal protein L31 type A / Ribosomal protein S16, conserved site / Ribosomal protein S16 signature. / Ribosomal protein L31 signature. / Ribosomal protein L31 / Ribosomal protein L31 superfamily / Ribosomal protein L31 / 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 S11, bacterial-type / Ribosomal protein S13, bacterial-type / Ribosomal protein S20 / 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 S18, conserved site / Ribosomal protein S18 signature. / L28p-like / 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 S10, conserved site / : / K Homology domain, type 2 / Ribosomal protein S3, C-terminal / Ribosomal protein S3, C-terminal domain superfamily / Ribosomal protein S15/S19, conserved site / KH domain / Ribosomal protein S19/S15 / Ribosomal protein S19/S15, superfamily / Ribosomal protein S10 / Ribosomal protein S3, C-terminal domain / Ribosomal protein S3 signature. / Ribosomal protein S10 signature. / Ribosomal protein S7, conserved site / Ribosomal protein S17, conserved site / K homology domain superfamily, prokaryotic type / Ribosomal protein S19 / Ribosomal protein S13, conserved site / Ribosomal protein S13 / 30s ribosomal protein S13, C-terminal / Ribosomal protein S4/S9 N-terminal domain / Ribosomal protein S14 / Ribosomal protein S4/S9, N-terminal / Ribosomal protein S4, conserved site / Type-2 KH domain profile. / Ribosomal protein S4/S9 N-terminal domain / Ribosomal protein S13/S18 / Ribosomal protein S4/S9 / Ribosomal protein S19 signature. / K homology domain-like, alpha/beta / Ribosomal protein S14p/S29e / Ribosomal protein S8 / Ribosomal protein S8 superfamily / Ribosomal protein S5, N-terminal, conserved site / Ribosomal protein S5 signature. / Ribosomal protein S5 / Ribosomal protein S5, N-terminal / Ribosomal S11, conserved site / Ribosomal protein S7 signature. / Ribosomal protein S10p/S20e / Ribosomal protein S13-like, H2TH / S5 double stranded RNA-binding domain profile. / Ribosomal protein S5, C-terminal / Ribosomal protein S9, conserved site / Ribosomal protein S5, N-terminal domain / Ribosomal protein S8 / Ribosomal protein S10 domain / Ribosomal protein S10 domain superfamily / Ribosomal protein S17 signature. / Ribosomal protein S5, C-terminal domain
Similarity search - Domain/homology
Small ribosomal subunit protein bS6 / Conserved domain protein / Small ribosomal subunit protein uS12 / Small ribosomal subunit protein uS7 / Small ribosomal subunit protein uS10 / Small ribosomal subunit protein uS19 / Small ribosomal subunit protein uS3 / Small ribosomal subunit protein uS17 / Small ribosomal subunit protein uS8 / Small ribosomal subunit protein uS5 ...Small ribosomal subunit protein bS6 / Conserved domain protein / Small ribosomal subunit protein uS12 / Small ribosomal subunit protein uS7 / Small ribosomal subunit protein uS10 / Small ribosomal subunit protein uS19 / Small ribosomal subunit protein uS3 / Small ribosomal subunit protein uS17 / Small ribosomal subunit protein uS8 / Small ribosomal subunit protein uS5 / Small ribosomal subunit protein uS13 / Small ribosomal subunit protein uS11 / Small ribosomal subunit protein uS4 / Small ribosomal subunit protein uS9 / Small ribosomal subunit protein bS16 / Small ribosomal subunit protein uS15 / Sigma 54 modulation/S30EA ribosomal protein C-terminal domain-containing protein / Small ribosomal subunit protein bS20 / Large ribosomal subunit protein bL31 / Small ribosomal subunit protein uS14A / Small ribosomal subunit protein bS18B
Similarity search - Component
Biological speciesMycolicibacterium smegmatis MC2 155 (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.5 Å
AuthorsKumar N / Sharma S / Kaushal PS
Funding support India, 1 items
OrganizationGrant numberCountry
Science and Engineering Research Board (SERB) India
CitationJournal: Nat Commun / Year: 2024
Title: Cryo- EM structure of the mycobacterial 70S ribosome in complex with ribosome hibernation promotion factor RafH.
Authors: Niraj Kumar / Shivani Sharma / Prem S Kaushal /
Abstract: Ribosome hibernation is a key survival strategy bacteria adopt under environmental stress, where a protein, hibernation promotion factor (HPF), transitorily inactivates the ribosome. Mycobacterium ...Ribosome hibernation is a key survival strategy bacteria adopt under environmental stress, where a protein, hibernation promotion factor (HPF), transitorily inactivates the ribosome. Mycobacterium tuberculosis encounters hypoxia (low oxygen) as a major stress in the host macrophages, and upregulates the expression of RafH protein, which is crucial for its survival. The RafH, a dual domain HPF, an orthologue of bacterial long HPF (HPF), hibernates ribosome in 70S monosome form, whereas in other bacteria, the HPF induces 70S ribosome dimerization and hibernates its ribosome in 100S disome form. Here, we report the cryo- EM structure of M. smegmatis, a close homolog of M. tuberculosis, 70S ribosome in complex with the RafH factor at an overall 2.8 Å resolution. The N- terminus domain (NTD) of RafH binds to the decoding center, similarly to HPF NTD. In contrast, the C- terminus domain (CTD) of RafH, which is larger than the HPF CTD, binds to a distinct site at the platform binding center of the ribosomal small subunit. The two domain-connecting linker regions, which remain mostly disordered in earlier reported HPF structures, interact mainly with the anti-Shine Dalgarno sequence of the 16S rRNA.
History
DepositionSep 24, 2023-
Header (metadata) releaseFeb 28, 2024-
Map releaseFeb 28, 2024-
UpdateFeb 28, 2024-
Current statusFeb 28, 2024Processing site: PDBj / Status: Released

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Structure visualization

Supplemental images

emd_37564.png

Downloads & links

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Map

FileDownload / File: emd_37564.map.gz / Format: CCP4 / Size: 209.3 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationMycobacterium smegmatis 30S ribosome (body 2) of 70S ribosome, E t-RNA and RafH.
Voxel sizeX=Y=Z: 1.07 Å
Density
Contour LevelBy AUTHOR: 0.045
Minimum - Maximum-0.034604132 - 0.19737574
Average (Standard dev.)0.0010772251 (±0.008260641)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions380380380
Spacing380380380
CellA=B=C: 406.6 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: Mycobacterium smegmatis 30S ribosome (body 2) of 70S...

Fileemd_37564_half_map_1.map
AnnotationMycobacterium smegmatis 30S ribosome (body 2) of 70S ribosome, E t-RNA and RafH, half 1.
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: Mycobacterium smegmatis 30S ribosome (body 2) of 70S...

Fileemd_37564_half_map_2.map
AnnotationMycobacterium smegmatis 30S ribosome (body 2) of 70S ribosome, E t-RNA and RafH, half 2.
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : 70S Ribosome + RafH protein

EntireName: 70S Ribosome + RafH protein
Components
  • Complex: 70S Ribosome + RafH protein
    • Complex: 70S RibosomeRibosome
      • RNA: 16S rRNA
      • RNA: E -tRNA
      • Protein or peptide: 30S ribosomal protein S22
      • 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 S14ARibosome
      • 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 S18BRibosome
      • Protein or peptide: 30S ribosomal protein S19
      • Protein or peptide: 30S ribosomal protein S20
      • Protein or peptide: 50S ribosomal protein L31
    • Complex: RafH
      • Protein or peptide: Ribosome hibernation promotion factor RafH

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Supramolecule #1: 70S Ribosome + RafH protein

SupramoleculeName: 70S Ribosome + RafH protein / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155

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Supramolecule #2: 70S Ribosome

SupramoleculeName: 70S Ribosome / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1-#22

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Supramolecule #3: RafH

SupramoleculeName: RafH / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #23

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Macromolecule #1: 16S rRNA

MacromoleculeName: 16S rRNA / type: rna / ID: 1 / Number of copies: 1
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 491.662594 KDa
SequenceString: UUUGGAGAGU UUGAUCCUGG CUCAGGACGA ACGCUGGCGG CGUGCUUAAC ACAUGCAAGU CGAACGGAAA GGCCCUUUCG GGGGUACUC GAGUGGCGAA CGGGUGAGUA ACACGUGGGU GAUCUGCCCU GCACUUUGGG AUAAGCCUGG GAAACUGGGU C UAAUACCG ...String:
UUUGGAGAGU UUGAUCCUGG CUCAGGACGA ACGCUGGCGG CGUGCUUAAC ACAUGCAAGU CGAACGGAAA GGCCCUUUCG GGGGUACUC GAGUGGCGAA CGGGUGAGUA ACACGUGGGU GAUCUGCCCU GCACUUUGGG AUAAGCCUGG GAAACUGGGU C UAAUACCG AAUACACCCU GCUGGUCGCA UGGCCUGGUA GGGGAAAGCU UUUGCGGUGU GGGAUGGGCC CGCGGCCUAU CA GCUUGUU GGUGGGGUGA UGGCCUACCA AGGCGACGAC GGGUAGCCGG CCUGAGAGGG UGACCGGCCA CACUGGGACU GAG AUACGG CCCAGACUCC UACGGGAGGC AGCAGUGGGG AAUAUUGCAC AAUGGGCGCA AGCCUGAUGC AGCGACGCCG CGUG AGGGA UGACGGCCUU CGGGUUGUAA ACCUCUUUCA GCACAGACGA AGCGCAAGUG ACGGUAUGUG CAGAAGAAGG ACCGG CCAA CUACGUGCCA GCAGCCGCGG UAAUACGUAG GGUCCGAGCG UUGUCCGGAA UUACUGGGCG UAAAGAGCUC GUAGGU GGU UUGUCGCGUU GUUCGUGAAA ACUCACAGCU UAACUGUGGG CGUGCGGGCG AUACGGGCAG ACUAGAGUAC UGCAGGG GA GACUGGAAUU CCUGGUGUAG CGGUGGAAUG CGCAGAUAUC AGGAGGAACA CCGGUGGCGA AGGCGGGUCU CUGGGCAG U AACUGACGCU GAGGAGCGAA AGCGUGGGGA GCGAACAGGA UUAGAUACCC UGGUAGUCCA CGCCGUAAAC GGUGGGUAC UAGGUGUGGG UUUCCUUCCU UGGGAUCCGU GCCGUAGCUA ACGCAUUAAG UACCCCGCCU GGGGAGUACG GCCGCAAGGC UAAAACUCA AAGGAAUUGA CGGGGGCCCG CACAAGCGGC GGAGCAUGUG GAUUAAUUCG AUGCAACGCG AAGAACCUUA C CUGGGUUU GACAUGCACA GGACGCCGGC AGAGAUGUCG GUUCCCUUGU GGCCUGUGUG CAGGUGGUGC AUGGCUGUCG UC AGCUCGU GUCGUGAGAU GUUGGGUUAA GUCCCGCAAC GAGCGCAACC CUUGUCUCAU GUUGCCAGCA CGUUAUGGUG GGG ACUCGU GAGAGACUGC CGGGGUCAAC UCGGAGGAAG GUGGGGAUGA CGUCAAGUCA UCAUGCCCCU UAUGUCCAGG GCUU CACAC AUGCUACAAU GGCCGGUACA AAGGGCUGCG AUGCCGUGAG GUGGAGCGAA UCCUUUCAAA GCCGGUCUCA GUUCG GAUC GGGGUCUGCA ACUCGACCCC GUGAAGUCGG AGUCGCUAGU AAUCGCAGAU CAGCAACGCU GCGGUGAAUA CGUUCC CGG GCCUUGUACA CACCGCCCGU CACGUCAUGA AAGUCGGUAA CACCCGAAGC CGGUGGCCUA ACCCUUGUGG AGGGAGC CG UCGAAGGUGG GAUCGGCGAU UGGGACGAAG UCGUAACAAG GUAGCCGUAC CGGAAGGUGC GGCUGGAUCA CCUC

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Macromolecule #2: E -tRNA

MacromoleculeName: E -tRNA / type: rna / ID: 2 / Number of copies: 1
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 24.465602 KDa
SequenceString:
CGCGGGGUGG AGCAGCCGGU AGCUCGUCUG GCUCAUAACC AGAAGAUCGU CGGUUCAAAU CCGGCCCCCG CAACCA

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Macromolecule #3: 30S ribosomal protein S22

MacromoleculeName: 30S ribosomal protein S22 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 4.1643 KDa
SequenceString:
MGSVIKKRRK RMSKKKHRKL LRRTRVQRRK LGK

UniProtKB: Conserved domain protein

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Macromolecule #4: 30S ribosomal protein S3

MacromoleculeName: 30S ribosomal protein S3 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 30.191227 KDa
SequenceString: MGQKINPHGF RLGITTEWKS RWYADKQYKD YVKEDVAIRK LLATGLERAG IADVEIERTR DRVRVDIHTA RPGIVIGRRG TEADRIRAD LEKLTGKQVQ LNILEVKNPE SQAQLVAQGV AEQLSNRVAF RRAMRKAIQS AMRQPNVKGI RVQCSGRLGG A EMSRSEFY ...String:
MGQKINPHGF RLGITTEWKS RWYADKQYKD YVKEDVAIRK LLATGLERAG IADVEIERTR DRVRVDIHTA RPGIVIGRRG TEADRIRAD LEKLTGKQVQ LNILEVKNPE SQAQLVAQGV AEQLSNRVAF RRAMRKAIQS AMRQPNVKGI RVQCSGRLGG A EMSRSEFY REGRVPLHTL RADIDYGLYE AKTTFGRIGV KVWIYKGDIV GGKRELAAAA PASDRPRRER PSGTRPRRSG SA GTTATST EAGRAATSDA PAAGTAAAAE APAESTES

UniProtKB: Small ribosomal subunit protein uS3

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Macromolecule #5: 30S ribosomal protein S4

MacromoleculeName: 30S ribosomal protein S4 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 23.415787 KDa
SequenceString: MARYTGPATR KSRRLGVDLV GGDQSFEKRP YPPGQHGRAR IKESEYRQQL QEKQKARFSY GVMEKQFRRY YEEANRQPGK TGDNLLRIL ESRLDNVVYR AGLARTRRMA RQLVSHGHFL VNGVKVDIPS YRVSQYDIID VKEKSLNTLP FQIARETAGE R PIPSWLQV ...String:
MARYTGPATR KSRRLGVDLV GGDQSFEKRP YPPGQHGRAR IKESEYRQQL QEKQKARFSY GVMEKQFRRY YEEANRQPGK TGDNLLRIL ESRLDNVVYR AGLARTRRMA RQLVSHGHFL VNGVKVDIPS YRVSQYDIID VKEKSLNTLP FQIARETAGE R PIPSWLQV VGERQRILVH QLPERAQIDV PLTEQLIVEL YSK

UniProtKB: Small ribosomal subunit protein uS4

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Macromolecule #6: 30S ribosomal protein S5

MacromoleculeName: 30S ribosomal protein S5 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 21.94609 KDa
SequenceString: MAEQAGAGSA QDNRGGRGRR DDRGGRGRDG GDKSNYIERV VSINRVSKVV KGGRRFSFTA LVIVGDGKGM VGVGYGKAKE VPAAIAKGV EEARKNFFRV PLIGSTITHP VQGEAAAGVV MLRPASPGTG VIAGGAARAV LECAGVHDIL AKSLGSDNAI N VVHATVAA ...String:
MAEQAGAGSA QDNRGGRGRR DDRGGRGRDG GDKSNYIERV VSINRVSKVV KGGRRFSFTA LVIVGDGKGM VGVGYGKAKE VPAAIAKGV EEARKNFFRV PLIGSTITHP VQGEAAAGVV MLRPASPGTG VIAGGAARAV LECAGVHDIL AKSLGSDNAI N VVHATVAA LKLLQRPEEV AARRGLPIED VAPAGMLKAR RESEALAAAA AREGSA

UniProtKB: Small ribosomal subunit protein uS5

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Macromolecule #7: 30S ribosomal protein S6

MacromoleculeName: 30S ribosomal protein S6 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 10.991637 KDa
SequenceString:
MRPYEIMVIL DPTLDERTVA PSLETFLNVI RKDGGTVDKV DIWGRRRLAY EIAKHAEGIY AVIDVKAEPA TVSELDRQLN LNESVLRTK VLRTDKH

UniProtKB: Small ribosomal subunit protein bS6

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Macromolecule #8: 30S ribosomal protein S7

MacromoleculeName: 30S ribosomal protein S7 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 17.660375 KDa
SequenceString:
MPRKGPAPKR PLVNDPVYGS QLVTQLVNKV LLEGKKSLAE RIVYGALEQA REKTGTDPVV TLKRALDNVK PALEVRSRRV GGATYQVPV EVRPDRSTTL ALRWLVNFSR QRREKTMVER LANEILDASN GLGASVKRRE DTHKMAEANR AFAHYRW

UniProtKB: Small ribosomal subunit protein uS7

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Macromolecule #9: 30S ribosomal protein S8

MacromoleculeName: 30S ribosomal protein S8 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 14.492638 KDa
SequenceString:
MTMTDPIADF LTRLRNANSA YHDEVTLPHS KLKANIAEIL KREGYISDYR TEDARVGKSL VVQLKYGPSR ERSIAGLRRV SKPGLRVYA KSTNLPRVLG GLGVAIISTS SGLLTDRQAA RQGVGGEVLA YVW

UniProtKB: Small ribosomal subunit protein uS8

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Macromolecule #10: 30S ribosomal protein S9

MacromoleculeName: 30S ribosomal protein S9 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 16.794365 KDa
SequenceString:
MTDVTETEVV TESAEPREPV IIDRPIQTVG RRKEAVVRVR LVPGTGQFNL DGRTLENYFP NKVHQQLIKA PLVTVDRVDQ FDIYAHLDG GGPSGQAGAL RLAIARALIL VQPEDRPALK KAGFLTRDPR AIERKKYGLK KARKAPQYSK R

UniProtKB: Small ribosomal subunit protein uS9

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Macromolecule #11: 30S ribosomal protein S10

MacromoleculeName: 30S ribosomal protein S10 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 11.454313 KDa
SequenceString:
MAGQKIRIRL KAYDHEAIDA SARKIVETVT RTGASVVGPV PLPTEKNVYC VIRSPHKYKD SREHFEMRTH KRLIDILDPT PKTVDALMR IDLPASVDVN IQ

UniProtKB: Small ribosomal subunit protein uS10

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Macromolecule #12: 30S ribosomal protein S11

MacromoleculeName: 30S ribosomal protein S11 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 14.671762 KDa
SequenceString:
MAQAKKGGTA AKKGQKTRRR EKKNVPHGAA HIKSTFNNTI VSITDPQGNV IAWASSGHVG FKGSRKSTPF AAQLAAENAA RKAQEHGVK KVDVFVKGPG SGRETAIRSL QAAGLEVGTI SDVTPQPHNG CRPPKRRRV

UniProtKB: Small ribosomal subunit protein uS11

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Macromolecule #13: 30S ribosomal protein S12

MacromoleculeName: 30S ribosomal protein S12 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 13.896366 KDa
SequenceString:
MPTIQQLVRK GRRDKIAKVK TAALKGSPQR RGVCTRVYTT TPKKPNSALR KVARVKLTSQ VEVTAYIPGE GHNLQEHSMV LVRGGRVKD LPGVRYKIIR GSLDTQGVKN RKQARSRYGA KKEKS

UniProtKB: Small ribosomal subunit protein uS12

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Macromolecule #14: 30S ribosomal protein S13

MacromoleculeName: 30S ribosomal protein S13 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 14.249619 KDa
SequenceString:
MARLVGVDLP RDKRMEIALT YIYGIGRTRS NEILAATGID KNMRTKDLTD DQVTVLRDYI EGNLKVEGDL RREVQADIRR KIEIGCYQG LRHRRGLPVR GQRTKTNART RKGPKRTIAG KKKAR

UniProtKB: Small ribosomal subunit protein uS13

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Macromolecule #15: 30S ribosomal protein S14A

MacromoleculeName: 30S ribosomal protein S14A / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 11.792728 KDa
SequenceString:
MAKKSKIVKN EQRRELVQRY AERRAELKRT IRDPASSPER RAAAVSALQR LPRDSSPVRL RNRDVVDGRP RGHLRKFGLS RVRVREMAH RGELPGVRKA SW

UniProtKB: Small ribosomal subunit protein uS14A

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Macromolecule #16: 30S ribosomal protein S15

MacromoleculeName: 30S ribosomal protein S15 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 10.368097 KDa
SequenceString:
MALTAEQKKE ILGQYGLHDT DTGSPEAQVA LLTKRIQDLT EHLKVHKHDH HSRRGLLLLV GRRRRLLKYV AQVDVARYRS LIERLGLRR

UniProtKB: Small ribosomal subunit protein uS15

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Macromolecule #17: 30S ribosomal protein S16

MacromoleculeName: 30S ribosomal protein S16 / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 16.795207 KDa
SequenceString:
MAVKIKLTRL GKIRNPQYRI IVADARTRRD GRAIEVIGRY HPKEEPSLIQ IDSERAQYWL GVGAQPTEPV LALLKITGDW QKFKGLPGA EGTLKVKEPK PSKLDLFNAA LAEAESGTTA AATTPKKKKA PKKDEAAEAP AEAAEAPAEA ADAASES

UniProtKB: Small ribosomal subunit protein bS16

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Macromolecule #18: 30S ribosomal protein S17

MacromoleculeName: 30S ribosomal protein S17 / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 11.127002 KDa
SequenceString:
MADQKGPKYT PAAEKPRGRR KTAIGYVVSD KMQKTIVVEL EDRKSHPLYG KIIRTTKKVK AHDENGEAGI GDRVSLMETR PLSATKRWR LVEILEKAK

UniProtKB: Small ribosomal subunit protein uS17

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Macromolecule #19: 30S ribosomal protein S18B

MacromoleculeName: 30S ribosomal protein S18B / type: protein_or_peptide / ID: 19 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 9.524188 KDa
SequenceString:
MAKSNKRRPA PEKPVKTRKC VFCSKKGQTI DYKDTALLRT YISERGKIRA RRVTGNCVQH QRDIAVAVKN AREVALLPFG SSTR

UniProtKB: Small ribosomal subunit protein bS18B

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Macromolecule #20: 30S ribosomal protein S19

MacromoleculeName: 30S ribosomal protein S19 / type: protein_or_peptide / ID: 20 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 10.800602 KDa
SequenceString:
MPRSLKKGPF VDDHLLKKVD VQNEKNTKQV IKTWSRRSTI IPDFIGHTFA VHDGRKHVPV FVTEAMVGHK LGEFAPTRTF KGHIKDDRK SKRR

UniProtKB: Small ribosomal subunit protein uS19

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Macromolecule #21: 30S ribosomal protein S20

MacromoleculeName: 30S ribosomal protein S20 / type: protein_or_peptide / ID: 21 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 9.556104 KDa
SequenceString:
MANIKSQIKR IRTNERRRLR NQSVKSSLRT AIRGFREAVD AGDKDKASEL LHATSRKLDK AASKGVIHPN QAANKKSALA LALNKL

UniProtKB: Small ribosomal subunit protein bS20

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Macromolecule #22: 50S ribosomal protein L31

MacromoleculeName: 50S ribosomal protein L31 / type: protein_or_peptide / ID: 22 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 8.312485 KDa
SequenceString:
MKTGIHPEYV DTTVQCGCGH SFTTRSTKQS GTIVVEVCSQ CHPFYTGKQK ILDSGGRVAR FEKRYGKRNK AAADK

UniProtKB: Large ribosomal subunit protein bL31

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Macromolecule #23: Ribosome hibernation promotion factor RafH

MacromoleculeName: Ribosome hibernation promotion factor RafH / type: protein_or_peptide / ID: 23 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 29.898936 KDa
Recombinant expressionOrganism: Escherichia coli BL21(DE3) (bacteria)
SequenceString: MDVDVSTDGE LPGAAEYARE KIGRLSRRAH RPVLHARVRL TRHGDPAVER PVIAQANLDV NGRQVRAQVE GVNAREAVDR LEARLRSRL ERIAEHWEAR RGGVPAEAGR EWRHESEPAR RPGYFPRPPE ERRIIRRKSF SMVPCTVDEA ALEMEMLDYD F HLFTEKGT ...String:
MDVDVSTDGE LPGAAEYARE KIGRLSRRAH RPVLHARVRL TRHGDPAVER PVIAQANLDV NGRQVRAQVE GVNAREAVDR LEARLRSRL ERIAEHWEAR RGGVPAEAGR EWRHESEPAR RPGYFPRPPE ERRIIRRKSF SMVPCTVDEA ALEMEMLDYD F HLFTEKGT GFAAVLYKGG PTGYRLVLVI PVPADELSPF EKPITISTHP APCLTQRDAV ERLGLLGLPF LFYIDAAEGR AS VIYRRYD GHYGLITPAD GHHHHHH

UniProtKB: Sigma 54 modulation/S30EA ribosomal protein C-terminal domain-containing protein

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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

Concentration1 mg/mL
BufferpH: 7.4 / Details: 20mM HEPES, 100mM NH4Cl, 20mM MgCl2, 3mM DTT,
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / 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: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.8 µm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Number grids imaged: 3 / Number real images: 12343 / Average exposure time: 2.0 sec. / Average electron dose: 1.34 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: NONE
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1.4)
Final 3D classificationSoftware - Name: RELION (ver. 3.1.4)
Final angle assignmentType: ANGULAR RECONSTITUTION / Software - Name: RELION (ver. 3.1.4)
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.5 Å / Resolution method: OTHER / Software - Name: RELION (ver. 3.1.4) / Details: Ad-hoc low pass filter, 3.5 Angstrom / Number images used: 44299
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelPDB ID:

8whx


Chain - Source name: PDB / Chain - Initial model type: experimental model
RefinementSpace: REAL / Protocol: RIGID BODY FIT
Output model

PDB-8wid:
Cryo- EM structure of Mycobacterium smegmatis 30S ribosomal subunit (body 2) of 70S ribosome, E- tRNA and RafH.

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