7egq

Electron Microscopy
3.35Å resolution

Co-transcriptional capping machineries in SARS-CoV-2 RTC: Coupling of N7-methyltransferase and 3'-5' exoribonuclease with polymerase reveals mechanisms for capping and proofreading

Released:

Function and Biology Details

Reactions catalysed:
S-adenosyl-L-methionine + a 5'-(5'-triphosphoguanosine)-[mRNA] = S-adenosyl-L-homocysteine + a 5'-(N(7)-methyl 5'-triphosphoguanosine)-[mRNA]
S-adenosyl-L-methionine + a 5'-(N(7)-methyl 5'-triphosphoguanosine)-(ribonucleotide)-[mRNA] = S-adenosyl-L-homocysteine + a 5'-(N(7)-methyl 5'-triphosphoguanosine)-(2'-O-methyl-ribonucleotide)-[mRNA]
Nucleoside triphosphate + RNA(n) = diphosphate + RNA(n+1)
GTP + a 5'-diphospho-[mRNA] = diphosphate + a 5'-(5'-triphosphoguanosine)-[mRNA]
Thiol-dependent hydrolysis of ester, thioester, amide, peptide and isopeptide bonds formed by the C-terminal Gly of ubiquitin (a 76-residue protein attached to proteins as an intracellular targeting signal).
TSAVLQ-|-SGFRK-NH(2) and SGVTFQ-|-GKFKK the two peptides corresponding to the two self-cleavage sites of the SARS 3C-like proteinase are the two most reactive peptide substrates. The enzyme exhibits a strong preference for substrates containing Gln at P1 position and Leu at P2 position.
ATP + H(2)O = ADP + phosphate
Biochemical function:
Biological process:
Cellular component:
  • not assigned

Structure analysis Details

Assembly composition:
hetero 22-mer (preferred)
PDBe Complex ID:
PDB-CPX-145082 (preferred)
Entry contents:
7 distinct polypeptide molecules
2 distinct RNA molecules
Macromolecules (9 distinct):
RNA-directed RNA polymerase nsp12 Chains: A, N
Molecule details ›
Chains: A, N
Length: 932 amino acids
Theoretical weight: 106.78 KDa
Source organism: Severe acute respiratory syndrome coronavirus 2
Expression system: Escherichia coli BL21(DE3)
UniProt:
  • Canonical: P0DTD1 (Residues: 4393-5324; Coverage: 13%)
Gene names: 1a-1b, rep
Sequence domains:
Non-structural protein 8 Chains: B, D, O, Q
Molecule details ›
Chains: B, D, O, Q
Length: 198 amino acids
Theoretical weight: 21.9 KDa
Source organism: Severe acute respiratory syndrome coronavirus 2
Expression system: Escherichia coli BL21(DE3)
UniProt:
  • Canonical: P0DTD1 (Residues: 3943-4140; Coverage: 3%)
Gene names: 1a-1b, rep
Sequence domains: Coronavirus replicase NSP8
Non-structural protein 7 Chains: C, P
Molecule details ›
Chains: C, P
Length: 83 amino acids
Theoretical weight: 9.25 KDa
Source organism: Severe acute respiratory syndrome coronavirus 2
Expression system: Escherichia coli BL21(DE3)
UniProt:
  • Canonical: P0DTD1 (Residues: 3860-3942; Coverage: 1%)
Gene names: 1a-1b, rep
Sequence domains: Coronavirus replicase NSP7
Helicase nsp13 Chains: E, F, R, S
Molecule details ›
Chains: E, F, R, S
Length: 601 amino acids
Theoretical weight: 66.93 KDa
Source organism: Severe acute respiratory syndrome coronavirus 2
Expression system: Escherichia coli BL21(DE3)
UniProt:
  • Canonical: P0DTD1 (Residues: 5325-5925; Coverage: 9%)
Gene names: 1a-1b, rep
Sequence domains:
Viral protein genome-linked nsp9 Chains: G, T
Molecule details ›
Chains: G, T
Length: 117 amino acids
Theoretical weight: 12.79 KDa
Source organism: Severe acute respiratory syndrome coronavirus 2
Expression system: Escherichia coli BL21(DE3)
UniProt:
  • Canonical: P0DTD1 (Residues: 4141-4253; Coverage: 2%)
Gene names: 1a-1b, rep
Sequence domains: Coronavirus replicase NSP9
Non-structural protein 10 Chains: H, U
Molecule details ›
Chains: H, U
Length: 139 amino acids
Theoretical weight: 14.8 KDa
Source organism: Severe acute respiratory syndrome coronavirus 2
Expression system: Escherichia coli BL21(DE3)
UniProt:
  • Canonical: P0DTD1 (Residues: 4254-4392; Coverage: 2%)
Gene names: 1a-1b, rep
Sequence domains: Coronavirus RNA synthesis protein NSP10
Guanine-N7 methyltransferase nsp14 Chains: K, X
Molecule details ›
Chains: K, X
Length: 527 amino acids
Theoretical weight: 59.89 KDa
Source organism: Severe acute respiratory syndrome coronavirus 2
Expression system: Escherichia coli BL21(DE3)
UniProt:
  • Canonical: P0DTD1 (Residues: 5926-6452; Coverage: 7%)
Gene names: 1a-1b, rep
Sequence domains: Coronavirus proofreading exoribonuclease
primer RNA Chains: I, L
Molecule details ›
Chains: I, L
Length: 25 nucleotides
Theoretical weight: 8.17 KDa
Template RNA Chains: J, M
Molecule details ›
Chains: J, M
Length: 33 nucleotides
Theoretical weight: 10.44 KDa

Ligands and Environments

2 bound ligands:
No modified residues

Experiments and Validation Details

Entry percentile scores
Resolution: 3.35Å
Relevant EMDB volumes: EMD-31138
Expression system: Escherichia coli BL21(DE3)