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- PDB-5u4i: Structural Basis of Co-translational Quality Control by ArfA and ... -
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Basic information
Entry | Database: PDB / ID: 5u4i | ||||||
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Title | Structural Basis of Co-translational Quality Control by ArfA and RF2 Bound to Ribosome | ||||||
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![]() | RIBOSOME / ArfA / RF2 / nonstop translation | ||||||
Function / homology | ![]() translation release factor activity, codon specific / stringent response / ribosomal large subunit binding / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / positive regulation of ribosome biogenesis ...translation release factor activity, codon specific / stringent response / ribosomal large subunit binding / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / positive regulation of ribosome biogenesis / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / translational termination / negative regulation of cytoplasmic translation / four-way junction DNA binding / DnaA-L2 complex / translation repressor activity / rescue of stalled ribosome / negative regulation of translational initiation / regulation of mRNA stability / negative regulation of DNA-templated DNA replication initiation / mRNA regulatory element binding translation repressor activity / assembly of large subunit precursor of preribosome / positive regulation of RNA splicing / ribosome assembly / transcription elongation factor complex / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / DNA endonuclease activity / response to reactive oxygen species / transcription antitermination / regulation of cell growth / DNA-templated transcription termination / maintenance of translational fidelity / response to radiation / mRNA 5'-UTR binding / regulation of translation / ribosome biogenesis / large ribosomal subunit / ribosome binding / transferase activity / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / small ribosomal subunit rRNA binding / 5S rRNA binding / ribosomal large subunit assembly / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / tRNA binding / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / viral translational frameshifting / response to antibiotic / negative regulation of DNA-templated transcription / mRNA binding / DNA binding / RNA binding / zinc ion binding / membrane / cytosol / cytoplasm Similarity search - Function | ||||||
Biological species | ![]() ![]() | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.5 Å | ||||||
![]() | Zeng, F. / Chen, Y. / Remis, J. / Shekhar, M. / Phillips, J.C. / Tajkhorshid, E. / Jin, H. | ||||||
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![]() | ![]() Title: Structural basis of co-translational quality control by ArfA and RF2 bound to ribosome. Authors: Fuxing Zeng / Yanbo Chen / Jonathan Remis / Mrinal Shekhar / James C Phillips / Emad Tajkhorshid / Hong Jin / ![]() Abstract: Quality control mechanisms intervene appropriately when defective translation events occur, in order to preserve the integrity of protein synthesis. Rescue of ribosomes translating on messenger RNAs ...Quality control mechanisms intervene appropriately when defective translation events occur, in order to preserve the integrity of protein synthesis. Rescue of ribosomes translating on messenger RNAs that lack stop codons is one of the co-translational quality control pathways. In many bacteria, ArfA recognizes stalled ribosomes and recruits the release factor RF2, which catalyses the termination of protein synthesis. Although an induced-fit mechanism of nonstop mRNA surveillance mediated by ArfA and RF2 has been reported, the molecular interaction between ArfA and RF2 in the ribosome that is responsible for the mechanism is unknown. Here we report an electron cryo-microscopy structure of ArfA and RF2 in complex with the 70S ribosome bound to a nonstop mRNA. The structure, which is consistent with our kinetic and biochemical data, reveals the molecular interactions that enable ArfA to specifically recruit RF2, not RF1, into the ribosome and to enable RF2 to release the truncated protein product in this co-translational quality control pathway. The positively charged C-terminal domain of ArfA anchors in the mRNA entry channel of the ribosome. Furthermore, binding of ArfA and RF2 induces conformational changes in the ribosomal decoding centre that are similar to those seen in other protein-involved decoding processes. Specific interactions between residues in the N-terminal domain of ArfA and RF2 help RF2 to adopt a catalytically competent conformation for peptide release. Our findings provide a framework for understanding recognition of the translational state of the ribosome by new proteins, and expand our knowledge of the decoding potential of the ribosome. | ||||||
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Structure visualization
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PDBx/mmCIF format | ![]() | 3.7 MB | Display | ![]() |
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-Validation report
Summary document | ![]() | 2 MB | Display | ![]() |
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Full document | ![]() | 2.1 MB | Display | |
Data in XML | ![]() | 222.7 KB | Display | |
Data in CIF | ![]() | 380.7 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8505MC ![]() 8506C ![]() 5u4jC C: citing same article ( M: map data used to model this data |
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Similar structure data |
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Assembly
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Components
-RNA chain , 5 types, 6 molecules ABaxyz
#1: RNA chain | Mass: 941761.625 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() ![]() | ||
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#2: RNA chain | Mass: 38177.762 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() ![]() | ||
#32: RNA chain | Mass: 497044.750 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() ![]() | ||
#55: RNA chain | Mass: 24802.785 Da / Num. of mol.: 2 / Source method: obtained synthetically / Details: fMet tRNA was in vitro transcribed / Source: (synth.) ![]() ![]() #56: RNA chain | | Mass: 5922.644 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) ![]() ![]() |
+50S ribosomal protein ... , 29 types, 29 molecules CDEFGHJKLMNOPQRSTUVWXYZ012345
-30S ribosomal protein ... , 20 types, 20 molecules bcdefghijklmnopqrstu
#33: Protein | Mass: 26781.670 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsB b0169 JW0164 / Source: (natural) ![]() ![]() |
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#34: Protein | Mass: 26031.316 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsC b3314 JW3276 / Source: (natural) ![]() ![]() |
#35: Protein | Mass: 23514.199 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsD ramA b3296 JW3258 / Source: (natural) ![]() ![]() |
#36: Protein | Mass: 17629.398 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsE spc b3303 JW3265 / Source: (natural) ![]() ![]() |
#37: Protein | Mass: 15727.512 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsF b4200 JW4158 / Source: (natural) ![]() ![]() |
#38: Protein | Mass: 20055.156 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsG b3341 JW3303 / Source: (natural) ![]() ![]() |
#39: Protein | Mass: 14015.361 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsH b3306 JW3268 / Source: (natural) ![]() ![]() |
#40: Protein | Mass: 14886.270 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsI b3230 JW3199 / Source: (natural) ![]() ![]() |
#41: Protein | Mass: 11755.597 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsJ nusE b3321 JW3283 / Source: (natural) ![]() ![]() |
#42: Protein | Mass: 12487.200 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsK b3297 JW3259 / Source: (natural) ![]() ![]() |
#43: Protein | Mass: 13636.961 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsL strA b3342 JW3304 / Source: (natural) ![]() ![]() |
#44: Protein | Mass: 13128.467 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsM b3298 JW3260 / Source: (natural) ![]() ![]() |
#45: Protein | Mass: 11475.364 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsN b3307 JW3269 / Source: (natural) ![]() ![]() |
#46: Protein | Mass: 10159.621 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsO secC b3165 JW3134 / Source: (natural) ![]() ![]() |
#47: Protein | Mass: 9207.572 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsP b2609 JW2590 / Source: (natural) ![]() ![]() |
#48: Protein | Mass: 9724.491 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsQ neaA b3311 JW3273 / Source: (natural) ![]() ![]() |
#49: Protein | Mass: 9063.508 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsR b4202 JW4160 / Source: (natural) ![]() ![]() |
#50: Protein | Mass: 9349.939 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsS b3316 JW3278 / Source: (natural) ![]() ![]() |
#51: Protein | Mass: 9577.268 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsT b0023 JW0022 / Source: (natural) ![]() ![]() |
#52: Protein | Mass: 8524.039 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Gene Name(s): rpsU b3065 JW3037 / Source: (natural) ![]() ![]() |
-Protein , 2 types, 2 molecules vw
#53: Protein | Mass: 43313.871 Da / Num. of mol.: 1 / Mutation: Q252MEQ Source method: isolated from a genetically manipulated source Details: Gene Name(s): prfB ECK2886 JW5847 supK Source: (gene. exp.) ![]() ![]() Strain: K12 / Gene: prfB, supK, b2891, JW5847 / Production host: ![]() ![]() |
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#54: Protein | Mass: 6513.396 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() Strain: K12 / Gene: arfA, yhdL, b4550, JW3253 / Production host: ![]() ![]() |
-Non-polymers , 2 types, 125 molecules 


#57: Chemical | ChemComp-MG / #58: Chemical | ChemComp-ZN / | |
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-Details
Has protein modification | Y |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
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