Literature summary extracted from

Urbonavicius, J.; Rutkiene, R.; Lopato, A.; Tauraite, D.; Stankeviciute, J.; Aucynaite, A.; Kaliniene, L.; van Tilbeurgh, H.; Meskys, R.
Evolution of tRNAPhe imG2 methyltransferases involved in the biosynthesis of wyosine derivatives in Archaea (2016), RNA, 22, 1871-1883 .

Cloned(Commentary)

EC Number	Cloned (Comment)	Organism
2.1.1.228	recombinant expression of His-tagged enzyme	Nanoarchaeum equitans
2.1.1.228	recombinant expression of His-tagged wild-type and mutant enzymes	Pyrococcus abyssi
2.1.1.282	recombinant expression of His-tagged enzyme	Nanoarchaeum equitans
2.1.1.282	recombinant expression of His-tagged enzyme	Saccharolobus solfataricus
2.1.1.282	recombinant expression of His-tagged wild-type and mutant enzymes	Pyrococcus abyssi

Protein Variants

EC Number	Protein Variants	Comment	Organism
2.1.1.228	E173A	site-directed mutagenesis, the mutant shows 9 and 26% of wild-type activity for imG and imG2 formation, respectively	Pyrococcus abyssi
2.1.1.228	E213A	site-directed mutagenesis, inactive mutant	Pyrococcus abyssi
2.1.1.228	F165A	site-directed mutagenesis, inactive mutant	Pyrococcus abyssi
2.1.1.228	P260N	site-directed mutagenesis, the mutant shows no and 114% of wild-type activity for imG and imG2 formation, respectively	Pyrococcus abyssi
2.1.1.228	P262A	site-directed mutagenesis, the mutant shows 5 and 8% of wild-type activity for imG and imG2 formation, respectively	Pyrococcus abyssi
2.1.1.228	R134A	site-directed mutagenesis, the mutant shows 2 and 4% of wild-type activity for imG and imG2 formation, respectively	Pyrococcus abyssi
2.1.1.228	R174A	site-directed mutagenesis, the mutant shows 8 and 69% of wild-type activity for imG and imG2 formation, respectively	Pyrococcus abyssi
2.1.1.282	E173A	site-directed mutagenesis, the mutant shows 9 and 26% of wild-type activity for imG and imG2 formation, respectively	Pyrococcus abyssi
2.1.1.282	E213A	site-directed mutagenesis, inactive mutant	Pyrococcus abyssi
2.1.1.282	F165A	site-directed mutagenesis, inactive mutant	Pyrococcus abyssi
2.1.1.282	P260N	site-directed mutagenesis, the mutant shows no and 114% of wild-type activity for imG and imG2 formation, respectively	Pyrococcus abyssi
2.1.1.282	P262A	site-directed mutagenesis, the mutant shows 5 and 8% of wild-type activity for imG and imG2 formation, respectively	Pyrococcus abyssi
2.1.1.282	R134A	site-directed mutagenesis, the mutant shows 2 and 4% of wild-type activity for imG and imG2 formation, respectively	Pyrococcus abyssi
2.1.1.282	R174A	site-directed mutagenesis, the mutant shows 8 and 69% of wild-type activity for imG and imG2 formation, respectively	Pyrococcus abyssi

Metals/Ions

EC Number	Metals/Ions	Comment	Organism
2.1.1.228	Mg2+	required	Pyrococcus abyssi
2.1.1.228	Mg2+	required	Nanoarchaeum equitans
2.1.1.282	Mg2+	required	Pyrococcus abyssi
2.1.1.282	Mg2+	required	Nanoarchaeum equitans
2.1.1.282	Mg2+	required	Saccharolobus solfataricus

Natural Substrates/ Products (Substrates)

EC Number	Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
2.1.1.228	S-adenosyl-L-methionine + 4-demethylwyosine	Pyrococcus abyssi	-	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 4-demethylwyosine	Nanoarchaeum equitans	-	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 4-demethylwyosine	Pyrococcus abyssi Orsay	-	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	Pyrococcus abyssi	-	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	Nanoarchaeum equitans	-	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	Pyrococcus abyssi Orsay	-	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	Pyrococcus abyssi	-	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.228	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	Nanoarchaeum equitans	-	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.228	S-adenosyl-L-methionine + guanine37 in tRNA	Pyrococcus abyssi	-	S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNA	-	?
2.1.1.228	S-adenosyl-L-methionine + guanine37 in tRNA	Nanoarchaeum equitans	-	S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNA	-	?
2.1.1.228	S-adenosyl-L-methionine + guanine37 in tRNA	Pyrococcus abyssi Orsay	-	S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNA	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	Pyrococcus abyssi	-	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	Nanoarchaeum equitans	-	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	Saccharolobus solfataricus	-	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	Saccharolobus solfataricus P2	-	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	Saccharolobus solfataricus JCM 11322	-	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	Saccharolobus solfataricus ATCC 35092	-	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	Saccharolobus solfataricus DSM 1617	-	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	Pyrococcus abyssi	-	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	Nanoarchaeum equitans	-	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	Pyrococcus abyssi	-	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	Nanoarchaeum equitans	-	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	Saccharolobus solfataricus	-	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	Saccharolobus solfataricus P2	-	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	Saccharolobus solfataricus JCM 11322	-	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	Saccharolobus solfataricus ATCC 35092	-	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	Saccharolobus solfataricus DSM 1617	-	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?

Organism

EC Number	Organism	UniProt	Comment	Textmining
2.1.1.228	Nanoarchaeum equitans	Q74NE4	-	-
2.1.1.228	Pyrococcus abyssi	Q9V2G1	-	-
2.1.1.228	Pyrococcus abyssi Orsay	Q9V2G1	-	-
2.1.1.282	Nanoarchaeum equitans	Q74NE4	-	-
2.1.1.282	Pyrococcus abyssi	Q9V2G1	-	-
2.1.1.282	Pyrococcus abyssi Orsay	Q9V2G1	-	-
2.1.1.282	Saccharolobus solfataricus	Q97W08	i.e. Sulfolobus solfataricus	-
2.1.1.282	Saccharolobus solfataricus ATCC 35092	Q97W08	i.e. Sulfolobus solfataricus	-
2.1.1.282	Saccharolobus solfataricus DSM 1617	Q97W08	i.e. Sulfolobus solfataricus	-
2.1.1.282	Saccharolobus solfataricus JCM 11322	Q97W08	i.e. Sulfolobus solfataricus	-
2.1.1.282	Saccharolobus solfataricus P2	Q97W08	i.e. Sulfolobus solfataricus	-

Purification (Commentary)

EC Number	Purification (Comment)	Organism
2.1.1.228	recombinant expression of His-tagged enzyme by affinity chromatography and desalting gel filtration	Nanoarchaeum equitans
2.1.1.228	recombinant expression of His-tagged wild-type and mutant enzymes to near homogeneity by 20 min heat treatment at 70°C and 65°C, respectively, followed by affinity chromatography and desalting gel filtration	Pyrococcus abyssi
2.1.1.282	recombinant expression of His-tagged enzyme by affinity chromatography and desalting gel filtration	Nanoarchaeum equitans
2.1.1.282	recombinant expression of His-tagged enzyme by affinity chromatography and desalting gel filtration	Saccharolobus solfataricus
2.1.1.282	recombinant expression of His-tagged wild-type and mutant enzymes to near homogeneity by 20 min heat treatment at 70°C and 65°C, respectively, followed by affinity chromatography and desalting gel filtration	Pyrococcus abyssi

Substrates and Products (Substrate)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
2.1.1.228	additional information	Nanoarchaeum equitans NEQ228 protein displays a dual tRNAPhe:m1G/imG2 methyltransferase activity. Two different types of substrates are used: (1) bulk tRNA, isolated from Salmonella enterica trmDELTA27 mutant containing the unmodified G37 nucleotide leading tothe formation of pm1G, and (2) tRNA, which is isolated from the Saccharomes cerevisiae DELTAtyw2 mutant that contains the imG-14 wyosine derivative leading to formation of pimG2pA dinucleotide and to a lesser extent to pm1G, likely resulting from the small amounts of G37-containing tRNAPhe present in the bulk tRNA isolates from the Scetyw2 mutant	Nanoarchaeum equitans	?	-	-
2.1.1.228	additional information	Pyrococcus abyssi PAB2272 protein displays a dual tRNAPhe:m1G/imG2 methyltransferase activity. Two different types of substrates are used: (1) bulk tRNA, isolated from Salmonella enterica trmDELTA27 mutant containing the unmodified G37 nucleotide leading to the formation of pm1G, and (2) tRNA, which is isolated from the Saccharomes cerevisiae DELTAtyw2 mutant that contains the imG-14 wyosine derivative leading to formation of pimG2pA dinucleotide and to a lesser extent to pm1G, likely resulting from the small amounts of G37-containing tRNAPhe present in the bulk tRNA isolates from the Scetyw2 mutant	Pyrococcus abyssi	?	-	-
2.1.1.228	additional information	Pyrococcus abyssi PAB2272 protein displays a dual tRNAPhe:m1G/imG2 methyltransferase activity. Two different types of substrates are used: (1) bulk tRNA, isolated from Salmonella enterica trmDELTA27 mutant containing the unmodified G37 nucleotide leading to the formation of pm1G, and (2) tRNA, which is isolated from the Saccharomes cerevisiae DELTAtyw2 mutant that contains the imG-14 wyosine derivative leading to formation of pimG2pA dinucleotide and to a lesser extent to pm1G, likely resulting from the small amounts of G37-containing tRNAPhe present in the bulk tRNA isolates from the Scetyw2 mutant	Pyrococcus abyssi Orsay	?	-	-
2.1.1.228	S-adenosyl-L-methionine + 4-demethylwyosine	-	Pyrococcus abyssi	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 4-demethylwyosine	-	Nanoarchaeum equitans	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 4-demethylwyosine	i.e. im-G14, activity of EC 2.1.1.282	Pyrococcus abyssi	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 4-demethylwyosine	i.e. im-G14, activity of EC 2.1.1.282	Nanoarchaeum equitans	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 4-demethylwyosine	-	Pyrococcus abyssi Orsay	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 4-demethylwyosine	i.e. im-G14, activity of EC 2.1.1.282	Pyrococcus abyssi Orsay	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	-	Pyrococcus abyssi	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	-	Nanoarchaeum equitans	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	i.e. yW-86, activity of EC 2.1.1.228	Pyrococcus abyssi	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	i.e. yW-86, activity of EC 2.1.1.228	Nanoarchaeum equitans	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	-	Pyrococcus abyssi Orsay	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.228	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	-	Pyrococcus abyssi	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.228	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	-	Nanoarchaeum equitans	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.228	S-adenosyl-L-methionine + guanine37 in tRNA	-	Pyrococcus abyssi	S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNA	-	?
2.1.1.228	S-adenosyl-L-methionine + guanine37 in tRNA	-	Nanoarchaeum equitans	S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNA	-	?
2.1.1.228	S-adenosyl-L-methionine + guanine37 in tRNA	-	Pyrococcus abyssi Orsay	S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNA	-	?
2.1.1.282	additional information	Nanoarchaeum equitans NEQ228 protein displays a dual tRNAPhe:m1G/imG2 methyltransferase activity. Two different types of substrates are used: (1) bulk tRNA, isolated from Salmonella enterica trmDELTA27 mutant containing the unmodified G37 nucleotide leading to the formation of pm1G, and (2) tRNA, which is isolated from the Saccharomes cerevisiae DELTAtyw2 mutant that contains the imG-14 wyosine derivative leading to formation of pimG2pA dinucleotide and to a lesser extent to pm1G, likely resulting from the small amounts of G37-containing tRNAPhe present in the bulk tRNA isolates from the Scetyw2 mutant	Nanoarchaeum equitans	?	-	-
2.1.1.282	additional information	Pyrococcus abyssi PAB2272 protein displays a dual tRNAPhe:m1G/imG2 methyltransferase activity. Two different types of substrates are used: (1) bulk tRNA, isolated from Salmonella enterica trmDELTA27 mutant containing the unmodified G37 nucleotide leading to the formation of pm1G, and (2) tRNA, which is isolated from the Saccharomes cerevisiae DELTAtyw2 mutant that contains the imG-14 wyosine derivative leading to formation of pimG2pA dinucleotide and to a lesser extent to pm1G, likely resulting from the small amounts of G37-containing tRNAPhe present in the bulk tRNA isolates from the Scetyw2 mutant	Pyrococcus abyssi	?	-	-
2.1.1.282	additional information	Sulfolobus solfataricus SSO2439 protein exhibits a tRNAPhe:imG2, but not tRNAPhe:m1G (EC 2.1.1.228), methyltransferase activity. The enzyme SSO2439 shows incorporation of the [methyl-14C] group into bulk yeast tRNA isolated from Saccharomyces cerevisiae DELTAtyw2 (containing imG-14 in tRNAPhe), but not into that from the Salmonella enterica trmDELTA27 (containing G37) mutant	Saccharolobus solfataricus	?	-	-
2.1.1.282	additional information	Sulfolobus solfataricus SSO2439 protein exhibits a tRNAPhe:imG2, but not tRNAPhe:m1G (EC 2.1.1.228), methyltransferase activity. The enzyme SSO2439 shows incorporation of the [methyl-14C] group into bulk yeast tRNA isolated from Saccharomyces cerevisiae DELTAtyw2 (containing imG-14 in tRNAPhe), but not into that from the Salmonella enterica trmDELTA27 (containing G37) mutant	Saccharolobus solfataricus P2	?	-	-
2.1.1.282	additional information	Sulfolobus solfataricus SSO2439 protein exhibits a tRNAPhe:imG2, but not tRNAPhe:m1G (EC 2.1.1.228), methyltransferase activity. The enzyme SSO2439 shows incorporation of the [methyl-14C] group into bulk yeast tRNA isolated from Saccharomyces cerevisiae DELTAtyw2 (containing imG-14 in tRNAPhe), but not into that from the Salmonella enterica trmDELTA27 (containing G37) mutant	Saccharolobus solfataricus JCM 11322	?	-	-
2.1.1.282	additional information	Sulfolobus solfataricus SSO2439 protein exhibits a tRNAPhe:imG2, but not tRNAPhe:m1G (EC 2.1.1.228), methyltransferase activity. The enzyme SSO2439 shows incorporation of the [methyl-14C] group into bulk yeast tRNA isolated from Saccharomyces cerevisiae DELTAtyw2 (containing imG-14 in tRNAPhe), but not into that from the Salmonella enterica trmDELTA27 (containing G37) mutant	Saccharolobus solfataricus ATCC 35092	?	-	-
2.1.1.282	additional information	Sulfolobus solfataricus SSO2439 protein exhibits a tRNAPhe:imG2, but not tRNAPhe:m1G (EC 2.1.1.228), methyltransferase activity. The enzyme SSO2439 shows incorporation of the [methyl-14C] group into bulk yeast tRNA isolated from Saccharomyces cerevisiae DELTAtyw2 (containing imG-14 in tRNAPhe), but not into that from the Salmonella enterica trmDELTA27 (containing G37) mutant	Saccharolobus solfataricus DSM 1617	?	-	-
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	-	Pyrococcus abyssi	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	-	Nanoarchaeum equitans	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	-	Saccharolobus solfataricus	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	i.e. im-G14, activity of EC 2.1.1.282	Pyrococcus abyssi	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	i.e. im-G14, activity of EC 2.1.1.282	Nanoarchaeum equitans	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	i.e. im-G14, activity of EC 2.1.1.282	Saccharolobus solfataricus	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	-	Saccharolobus solfataricus P2	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	i.e. im-G14, activity of EC 2.1.1.282	Saccharolobus solfataricus P2	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	-	Saccharolobus solfataricus JCM 11322	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	i.e. im-G14, activity of EC 2.1.1.282	Saccharolobus solfataricus JCM 11322	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	-	Saccharolobus solfataricus ATCC 35092	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	i.e. im-G14, activity of EC 2.1.1.282	Saccharolobus solfataricus ATCC 35092	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	-	Saccharolobus solfataricus DSM 1617	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 4-demethylwyosine	i.e. im-G14, activity of EC 2.1.1.282	Saccharolobus solfataricus DSM 1617	S-adenosyl-L-homocysteine + isowyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	-	Pyrococcus abyssi	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	-	Nanoarchaeum equitans	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	i.e. yW-86, activity of EC 2.1.1.228	Pyrococcus abyssi	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-aminocarboxypropyl-demethylwyosine	i.e. yW-86, activity of EC 2.1.1.228	Nanoarchaeum equitans	S-adenosyl-L-homocysteine + wyosine	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	-	Pyrococcus abyssi	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	-	Nanoarchaeum equitans	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	-	Saccharolobus solfataricus	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	-	Saccharolobus solfataricus P2	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	-	Saccharolobus solfataricus JCM 11322	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	-	Saccharolobus solfataricus ATCC 35092	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?
2.1.1.282	S-adenosyl-L-methionine + 7-[(3S)-(3-amino-3-carboxypropyl)]-4-demethylwyosine37 in tRNAPhe	-	Saccharolobus solfataricus DSM 1617	S-adenosyl-L-homocysteine + 7-[(3S)-(3-amino-3-carboxypropyl)]wyosine37 in tRNAPhe	-	?

Synonyms

EC Number	Synonyms	Comment	Organism
2.1.1.228	aTrm5a methyltransferase	-	Pyrococcus abyssi
2.1.1.228	aTrm5a methyltransferase	-	Nanoarchaeum equitans
2.1.1.228	aTrm5a/Taw22-like enzyme	-	Pyrococcus abyssi
2.1.1.228	More	see also EC 2.1.1.282	Pyrococcus abyssi
2.1.1.228	More	see also EC 2.1.1.282	Nanoarchaeum equitans
2.1.1.228	NEQ228	-	Nanoarchaeum equitans
2.1.1.228	PAB2272	-	Pyrococcus abyssi
2.1.1.228	TAW22	-	Pyrococcus abyssi
2.1.1.228	TAW22	-	Nanoarchaeum equitans
2.1.1.228	Trm5a	-	Pyrococcus abyssi
2.1.1.228	Trm5a	-	Nanoarchaeum equitans
2.1.1.228	Trm5a/Taw22-like enzyme	-	Nanoarchaeum equitans
2.1.1.228	tRNAPhe:imG2 methyltransferase	-	Pyrococcus abyssi
2.1.1.228	tRNAPhe:imG2 methyltransferase	-	Nanoarchaeum equitans
2.1.1.282	aTrm5a methyltransferase	-	Pyrococcus abyssi
2.1.1.282	aTrm5a methyltransferase	-	Nanoarchaeum equitans
2.1.1.282	aTrm5a methyltransferase	-	Saccharolobus solfataricus
2.1.1.282	aTrm5a-type enzyme	-	Saccharolobus solfataricus
2.1.1.282	aTrm5a/Taw22-like enzyme	-	Pyrococcus abyssi
2.1.1.282	More	see also EC 2.1.1.228	Pyrococcus abyssi
2.1.1.282	More	see also EC 2.1.1.228	Nanoarchaeum equitans
2.1.1.282	NEQ228	-	Nanoarchaeum equitans
2.1.1.282	PAB2272	-	Pyrococcus abyssi
2.1.1.282	SSO2439	-	Saccharolobus solfataricus
2.1.1.282	Taw21	-	Saccharolobus solfataricus
2.1.1.282	TAW22	-	Pyrococcus abyssi
2.1.1.282	TAW22	-	Nanoarchaeum equitans
2.1.1.282	Trm5a	-	Pyrococcus abyssi
2.1.1.282	Trm5a	-	Nanoarchaeum equitans
2.1.1.282	Trm5a/Taw22-like enzyme	-	Nanoarchaeum equitans
2.1.1.282	tRNAPhe:imG2 methyltransferase	-	Pyrococcus abyssi
2.1.1.282	tRNAPhe:imG2 methyltransferase	-	Nanoarchaeum equitans
2.1.1.282	tRNAPhe:imG2 methyltransferase	-	Saccharolobus solfataricus

Temperature Optimum [°C]

EC Number	Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
2.1.1.228	50	-	assay at	Pyrococcus abyssi
2.1.1.228	50	-	assay at	Nanoarchaeum equitans
2.1.1.282	50	-	assay at	Pyrococcus abyssi
2.1.1.282	50	-	assay at	Nanoarchaeum equitans
2.1.1.282	50	-	assay at	Saccharolobus solfataricus

pH Optimum

EC Number	pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
2.1.1.228	8	-	assay at	Pyrococcus abyssi
2.1.1.228	8	-	assay at	Nanoarchaeum equitans
2.1.1.282	8	-	assay at	Pyrococcus abyssi
2.1.1.282	8	-	assay at	Nanoarchaeum equitans
2.1.1.282	8	-	assay at	Saccharolobus solfataricus

Cofactor

EC Number	Cofactor	Comment	Organism
2.1.1.228	S-adenosyl-L-methionine	-	Pyrococcus abyssi
2.1.1.228	S-adenosyl-L-methionine	-	Nanoarchaeum equitans
2.1.1.282	S-adenosyl-L-methionine	-	Pyrococcus abyssi
2.1.1.282	S-adenosyl-L-methionine	-	Nanoarchaeum equitans
2.1.1.282	S-adenosyl-L-methionine	-	Saccharolobus solfataricus

General Information

EC Number	General Information	Comment	Organism
2.1.1.228	evolution	archaeal Trm5a, a member of the archaeal Trm5a/b/c family of enzymes involved in the biosynthesis of the wyosine derivatives, division of the family aTrm5 into three subfamilies aTrm5a (further divided into Taw21 and Taw22 which are monofunctional and bifunctional aTrm5a), aTrm5b, and aTrm5c. While the enzymes belonging to these subfamilies do not significantly differ in their AdoMet-binding site, small differences have been observed within the NPPY motif, which, in certain amino-methyltransferases, is involved in the positioning of the target nitrogen atom. In contrast, the N-terminal sequences of the aforementioned enzymes differ substantially, e.g. a small conservative domain called D1 is present in aTrm5b and aTrm5c but absent in most of the aTrm5a proteins. Evolution of tRNAPhe:imG2 methyltransferases involved in the biosynthesis of wyosine derivatives in Archaea. Amino acid sequence alignment of Trm5a/b/c family of proteins. Monofunctional and bifunctional aTrm5a enzymes, overview	Nanoarchaeum equitans
2.1.1.228	evolution	archaeal Trm5a, a member of the archaeal Trm5a/b/c family of enzymes involved in the biosynthesis of the wyosine derivatives, division of the family aTrm5 into three subfamilies aTrm5a (further divided into Taw21 and Taw22 which are monofunctional and bifunctional aTrm5a), aTrm5b, and aTrm5c. While the enzymes belonging to these subfamilies do not significantly differ in their AdoMet-binding site, small differences have been observed within the NPPY motif, which, in certain amino-methyltransferases, is involved in the positioning of the target nitrogen atom. In contrast, the N-terminal sequences of the aforementioned enzymes differ substantially, e.g. a small conservative domain called D1 is present in aTrm5b and aTrm5c but absent in most of the aTrm5a proteins. Evolution of tRNAPhe:imG2 methyltransferases involved in the biosynthesis of wyosine derivatives in Archaea. Amino acid sequence alignment of Trm5a/b/c/ family of proteins. Monofunctional and bifunctional aTrm5a enzymes, overview	Pyrococcus abyssi
2.1.1.228	malfunction	substitutions of individual conservative amino acids of Pyrococcus abyssi Taw22 (P260N, E173A, and R174A) have a differential effect on the formation of m1G/imG2, while replacement of R134, F165, E213, and P262 with alanine abolishes the formation of both derivatives of G37	Pyrococcus abyssi
2.1.1.228	malfunction	substitutions of individual conservative amino acids of Pyrococcus abyssi Taw22 (P260N, E173A, and R174A) have a differential effect on the formation of m1G/imG2, while replacement of R134, F165, E213, and P262 with alanine abolishes the formation of both derivatives of G37	Nanoarchaeum equitans
2.1.1.228	metabolism	putative enzymatic pathway leading to the formation of wyosine derivatives in Archaea	Pyrococcus abyssi
2.1.1.228	metabolism	putative enzymatic pathway leading to the formation of wyosine derivatives in Archaea	Nanoarchaeum equitans
2.1.1.228	physiological function	tricyclic wyosine derivatives are found at position 37 of eukaryotic and archaeal tRNAPhe. In Archaea, the intermediate imG-14 is targeted by three different enzymes that catalyze the formation of yW-86, imG, and imG2. Methyltransferase aTrm5a/Taw22 likely catalyzes two distinct reactions: N1-methylation of guanosine to yield m1G (EC 2.1.1.228), and C7-methylation of imG-14 to yield imG2 (EC 2.1.1.282)	Pyrococcus abyssi
2.1.1.228	physiological function	tricyclic wyosine derivatives are found at position 37 of eukaryotic and archaeal tRNAPhe. In Archaea, the intermediate imG-14 is targeted by three different enzymes that catalyze the formation of yW-86, imG, and imG2. Methyltransferase aTrm5a/Taw22 likely catalyzes two distinct reactions: N1-methylation of guanosine to yield m1G (EC 2.1.1.228), and C7-methylation of imG-14 to yield imG2 (EC 2.1.1.282)	Nanoarchaeum equitans
2.1.1.282	evolution	archaeal Trm5a, a member of the archaeal Trm5a/b/c family of enzymes involved in the biosynthesis of the wyosine derivatives, division of the family aTrm5 into three subfamilies aTrm5a (further divided into Taw21 and Taw22 which are monofunctional and bifunctional aTrm5a ), aTrm5b, and aTrm5c. While the enzymes belonging to these subfamilies do not significantly differ in their AdoMet-binding site, small differences have been observed within the NPPY motif, which, in certain amino-methyltransferases, is involved in the positioning of the target nitrogen atom. In contrast, the N-terminal sequences of the aforementioned enzymes differ substantially, e.g. a small conservative domain called D1 is present in aTrm5b and aTrm5c but absent in most of the aTrm5a proteins. Evolution of tRNAPhe:imG2 methyltransferases involved in the biosynthesis of wyosine derivatives in Archaea. Amino acid sequence alignment of Trm5a/b/c/ family of proteins. Monofunctional and bifunctional aTrm5a enzymes, overview	Nanoarchaeum equitans
2.1.1.282	evolution	archaeal Trm5a, a member of the archaeal Trm5a/b/c family of enzymes involved in the biosynthesis of the wyosine derivatives, division of the family aTrm5 into three subfamilies aTrm5a (further divided into Taw21 and Taw22 which are monofunctional and bifunctional aTrm5a ), aTrm5b, and aTrm5c. While the enzymes belonging to these subfamilies do not significantly differ in their AdoMet-binding site, small differences have been observed within the NPPY motif, which, in certain amino-methyltransferases, is involved in the positioning of the target nitrogen atom. In contrast, the N-terminal sequences of the aforementioned enzymes differ substantially, e.g. a small conservative domain called D1 is present in aTrm5b and aTrm5c but absent in most of the aTrm5a proteins. Evolution of tRNAPhe:imG2 methyltransferases involved in the biosynthesis of wyosine derivatives in Archaea. Amino acid sequence alignment of Trm5a/b/c/ family of proteins. Monofunctional and bifunctional aTrm5a enzymes, overview. Crenarcheota Saccharolobus solfataricus as well as in other Sulfalobales and Desulfurococcales, two different tRNAPhe methyltransferases are involved in the biosynthesis of mimG, catalyzing the formation of m1G (Trm5c, EC 2.1.1.228) and imG2 (Trm5a, EC 2.1.1.282) at position 37 in tRNAPhe, respectively	Saccharolobus solfataricus
2.1.1.282	evolution	archaeal Trm5a, a member of the archaeal Trm5a/b/c family of enzymes involved in the biosynthesis of the wyosine derivatives, division of the family aTrm5 into three subfamilies aTrm5a (further divided into Taw21 and Taw22 which are monofunctional and bifunctional aTrm5a), aTrm5b, and aTrm5c. While the enzymes belonging to these subfamilies do not significantly differ in their AdoMet-binding site, small differences have been observed within the NPPY motif, which, in certain amino-methyltransferases, is involved in the positioning of the target nitrogen atom. In contrast, the N-terminal sequences of the aforementioned enzymes differ substantially, e.g. a small conservative domain called D1 is present in aTrm5b and aTrm5c but absent in most of the aTrm5a proteins. Evolution of tRNAPhe:imG2 methyltransferases involved in the biosynthesis of wyosine derivatives in Archaea. Amino acid sequence alignment of Trm5a/b/c/ family of proteins. Monofunctional and bifunctional aTrm5a enzymes, overview	Pyrococcus abyssi
2.1.1.282	malfunction	substitutions of individual conservative amino acids of Pyrococcus abyssi Taw22 (P260N, E173A, and R174A) have a differential effect on the formation of m1G/imG2, while replacement of R134, F165, E213, and P262 with alanine abolishes the formation of both derivatives of G37	Pyrococcus abyssi
2.1.1.282	malfunction	substitutions of individual conservative amino acids of Pyrococcus abyssi Taw22 (P260N, E173A, and R174A) have a differential effect on the formation of m1G/imG2, while replacement of R134, F165, E213, and P262 with alanine abolishes the formation of both derivatives of G37	Nanoarchaeum equitans
2.1.1.282	malfunction	substitutions of individual conservative amino acids of Pyrococcus abyssi Taw22 (P260N, E173A, and R174A) have a differential effect on the formation of m1G/imG2, while replacement of R134, F165, E213, and P262 with alanine abolishes the formation of both derivatives of G37	Saccharolobus solfataricus
2.1.1.282	metabolism	putative enzymatic pathway leading to the formation of wyosine derivatives in Archaea	Pyrococcus abyssi
2.1.1.282	metabolism	putative enzymatic pathway leading to the formation of wyosine derivatives in Archaea	Nanoarchaeum equitans
2.1.1.282	metabolism	putative enzymatic pathway leading to the formation of wyosine derivatives in Archaea	Saccharolobus solfataricus
2.1.1.282	physiological function	tricyclic wyosine derivatives are found at position 37 of eukaryotic and archaeal tRNAPhe. In Archaea, the intermediate imG-14 is targeted by three different enzymes that catalyze the formation of yW-86, imG, and imG2. Methyltransferase aTrm5a/Taw22 likely catalyzes two distinct reactions: N1-methylation of guanosine to yield m1G (EC 2.1.1.228), and C7-methylation of imG-14 to yield imG2 (EC 2.1.1.282)	Pyrococcus abyssi
2.1.1.282	physiological function	tricyclic wyosine derivatives are found at position 37 of eukaryotic and archaeal tRNAPhe. In Archaea, the intermediate imG-14 is targeted by three different enzymes that catalyze the formation of yW-86, imG, and imG2. Methyltransferase aTrm5a/Taw22 likely catalyzes two distinct reactions: N1-methylation of guanosine to yield m1G (EC 2.1.1.228), and C7-methylation of imG-14 to yield imG2 (EC 2.1.1.282)	Nanoarchaeum equitans
2.1.1.282	physiological function	tricyclic wyosine derivatives are found at position 37 of eukaryotic and archaeal tRNAPhe. In Archaea, the intermediate imG-14 is targeted by three different enzymes that catalyze the formation of yW-86, imG, and imG2. Methyltransferase aTrm5a/Taw22 likely catalyzes two distinct reactions: N1-methylation of guanosine to yield m1G (EC 2.1.1.228), and C7-methylation of imG-14 to yield imG2 (EC 2.1.1.282)	Saccharolobus solfataricus