Cloned (Comment) | Organism |
---|---|
recombinant coexpression of His-tagged TaArcS and His-tagged TkRaSEA in Escherichia coli | Thermococcus kodakarensis |
recombinant coexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain ER2566 | Methanosarcina acetivorans |
Protein Variants | Comment | Organism |
---|---|---|
C51A | site-directed mutagenesis of a residue from the radical SAM enzymes' conserved CX3CX2C motif. No T1 fragment containing G+ is detected, but signals of U(s4U)AACAAA(q0kN)CGp are observed in the T1 fragments of tRNACys from C51A-expressing cells | Methanosarcina acetivorans |
C59A | site-directed mutagenesis of a residue from the radical SAM enzymes' conserved CX3CX2C motif. No T1 fragment containing G+ is detected, but signals of U(s4U)AACAAA(q0kN)CGp are observed in the T1 fragments of tRNACys from C59A-expressing cells | Methanosarcina acetivorans |
C62A | site-directed mutagenesis of a residue from the radical SAM enzymes' conserved CX3CX2C motif. No T1 fragment containing G+ is detected, but signals of U(s4U)AACAAA(q0kN)CGp are observed in the T1 fragments of tRNACys from C62A-expressing cells | Methanosarcina acetivorans |
additional information | construction of a TK1135 disruption strain (DELTATK1135). The G+ peak disappears from the tRNA fraction of strain DELTATK1135 and a small q0kN peak is observed | Thermococcus kodakarensis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | in the [4Fe-4S] cluster | Methanosarcina acetivorans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | Thermococcus kodakarensis | reaction of subunit beta | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | Methanosarcina acetivorans | reaction of subunit beta | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | Methanosarcina acetivorans ATCC 35395 | reaction of subunit beta | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | Thermococcus kodakarensis JCM 12380 | reaction of subunit beta | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | Methanosarcina acetivorans DSM 2834 | reaction of subunit beta | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | Methanosarcina acetivorans JCM 12185 | reaction of subunit beta | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | Thermococcus kodakarensis ATCC BAA-918 | reaction of subunit beta | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanosarcina acetivorans | Q8THK1 | ArcS subunit beta | - |
Methanosarcina acetivorans ATCC 35395 | Q8THK1 | ArcS subunit beta | - |
Methanosarcina acetivorans DSM 2834 | Q8THK1 | ArcS subunit beta | - |
Methanosarcina acetivorans JCM 12185 | Q8THK1 | ArcS subunit beta | - |
Thermococcus kodakarensis | Q5JE80 | ArcS subunit beta | - |
Thermococcus kodakarensis ATCC BAA-918 | Q5JE80 | ArcS subunit beta | - |
Thermococcus kodakarensis JCM 12380 | Q5JE80 | ArcS subunit beta | - |
Purification (Comment) | Organism |
---|---|
copurification of recombinant His-tagged TaArcS and His-tagged TkRaSEA from Escherichia coli by nickel affinity chromatography | Thermococcus kodakarensis |
copurification of recombinant His-tagged wild-type and mutant enzyme complexes from Escherichia coli by nickel affinity chromatography | Methanosarcina acetivorans |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine = 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | reaction mechanism of RaSEA, overview | Thermococcus kodakarensis | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine = 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | reaction mechanism of RaSEA, overview | Methanosarcina acetivorans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | reaction of subunit beta | Thermococcus kodakarensis | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | reaction of subunit beta | Methanosarcina acetivorans | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | reaction of subunit beta. G+ is successfully synthesized in recombinant Escherichia coli cells by MaArcTGT and the MaArcS-MaRaSEA complex | Methanosarcina acetivorans | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | reaction of subunit beta | Methanosarcina acetivorans ATCC 35395 | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | reaction of subunit beta. G+ is successfully synthesized in recombinant Escherichia coli cells by MaArcTGT and the MaArcS-MaRaSEA complex | Methanosarcina acetivorans ATCC 35395 | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | reaction of subunit beta | Thermococcus kodakarensis JCM 12380 | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | reaction of subunit beta | Methanosarcina acetivorans DSM 2834 | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | reaction of subunit beta. G+ is successfully synthesized in recombinant Escherichia coli cells by MaArcTGT and the MaArcS-MaRaSEA complex | Methanosarcina acetivorans DSM 2834 | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | reaction of subunit beta | Methanosarcina acetivorans JCM 12185 | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | reaction of subunit beta. G+ is successfully synthesized in recombinant Escherichia coli cells by MaArcTGT and the MaArcS-MaRaSEA complex | Methanosarcina acetivorans JCM 12185 | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? | |
7-N-[(5S)-5-amino-5-carboxypentyl]formamidino-7-deazaguanosine15 in tRNA + S-adenosyl-L-methionine | reaction of subunit beta | Thermococcus kodakarensis ATCC BAA-918 | 5'-deoxyadenosine + archaeosine15 in tRNA + 2 H+ + L-1-piperideine-6-carboxylate + L-methionine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
archaeosine synthase subunit beta | - |
Thermococcus kodakarensis |
archaeosine synthase subunit beta | - |
Methanosarcina acetivorans |
MaRaSEA | - |
Methanosarcina acetivorans |
MA_4513 | - |
Methanosarcina acetivorans |
q0kN-tRNA lyase | - |
Thermococcus kodakarensis |
q0kN-tRNA lyase | - |
Methanosarcina acetivorans |
RaSEA | - |
Thermococcus kodakarensis |
RaSEA | - |
Methanosarcina acetivorans |
TK1135 | locus name of subunit beta | Thermococcus kodakarensis |
TkRaSEA | - |
Thermococcus kodakarensis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | subunit B is a SAM radical enzyme | Thermococcus kodakarensis | |
S-adenosyl-L-methionine | subunit B is a SAM radical enzyme | Methanosarcina acetivorans | |
[4Fe-4S] cluster | - |
Methanosarcina acetivorans |
General Information | Comment | Organism |
---|---|---|
evolution | the hyperthermophilic euryarchaeon Thermococcus kodakarensis lacks an arcTGT orthologue | Thermococcus kodakarensis |
evolution | the hyperthermophilic euryarchaeon Thermococcus kodakarensis lacks an arcTGT orthologue | Methanosarcina acetivorans |
malfunction | in a TK1135 disruption strain (DELTATK1135), the G+ peak disappears from the tRNA fraction and a small q0kN peak is observed | Thermococcus kodakarensis |
malfunction | in a TK1135 disruption strain (DELTATK1135), the G+ peak disappears from the tRNA fraction and a small q0kN peak is observed | Methanosarcina acetivorans |
metabolism | archaeosine biosynthesis pathway, overview | Thermococcus kodakarensis |
metabolism | archaeosine biosynthesis pathway, overview | Methanosarcina acetivorans |
additional information | the archaeosine synthases from Thermococcus kodakarensis is composed of ArcS (subunit alpha) and the radical SAM enzyme TkRaSEA (q0kN-tRNA lyase, subunit beta) both forming a robust complex, but the interaction between TkRaSEA and Thermoplasma acidophilum ArcS is considerably weaker. Interaction analysis, overview | Thermococcus kodakarensis |
additional information | the archaeosine synthases from Thermococcus kodakarensis is composed of ArcS (subunit alpha) and the radical SAM enzyme TkRaSEA (q0kN-tRNA lyase, subunit beta) both forming a robust complex, but the interaction between TkRaSEA and Thermoplasma acidophilum ArcS is considerably weaker. Interaction analysis, overview. Residues C51, C59 and C62 in the conserved CX3CX2C motif of radical SAM enzyme MaRaSEA are involved in G+ formation and coordinate iron atoms of the [4Fe-4S] cluster in MaRaSEA | Methanosarcina acetivorans |
physiological function | archaeosine (G+), 7-formamidino-7-deazaguanosine, is an archaea-specific modified nucleoside found at the 15th position of tRNAs. In Euryarchaeota, 7-cyano-7-deazaguanine (preQ0)-containing tRNA (q0N-tRNA), synthesized by archaeal tRNA-guanine transglycosylase (ArcTGT), is converted to G+-containing tRNA (G+-tRNA) by the paralogue of ArcTGT, ArcS. Several euryarchaeal ArcSs have lysine transfer activity to q0N-tRNA to form q0kN-tRNA, which has a preQ0 lysine adduct as a base. ArcS forms a robust complex with a radical S-adenosylmethionine (SAM) enzyme named RaSEA. The ArcS-RaSEA complex anaerobically converts q0N-tRNA to G+-tRNA in the presence of SAM and lysine via q0kN-tRNA. It is proposed that ArcS and RaSEA should be considered an archaeosine synthase alpha-subunit (lysine transferase) and beta-subunit (q0kN-tRNA lyase), respectively. The TK1135 product is involved in G+ biosynthesis and q0kN is an intermediate in the synthesis of G+ | Thermococcus kodakarensis |
physiological function | archaeosine (G+), 7-formamidino-7-deazaguanosine, is an archaea-specific modified nucleoside found at the 15th position of tRNAs. In Euryarchaeota, 7-cyano-7-deazaguanine (preQ0)-containing tRNA (q0N-tRNA), synthesized by archaeal tRNA-guanine transglycosylase (ArcTGT), is converted to G+-containing tRNA (G+-tRNA) by the paralogue of ArcTGT, ArcS. Several euryarchaeal ArcSs have lysine transfer activity to q0N-tRNA to form q0kN-tRNA, which has a preQ0 lysine adduct as a base. ArcS forms a robust complex with a radical S-adenosylmethionine (SAM) enzyme named RaSEA. The ArcS-RaSEA complex anaerobically converts q0N-tRNA to G+-tRNA in the presence of SAM and lysine via q0kN-tRNA. It is proposed that ArcS and RaSEA should be considered an archaeosine synthase alpha-subunit (lysine transferase) and beta-subunit (q0kN-tRNA lyase), respectively. The TK1135 product is involved in G+ biosynthesis and q0kN is an intermediate in the synthesis of G+ | Methanosarcina acetivorans |