Any feedback?
Enzyme Nomenclature
Information on EC 2.8.1.15 - tRNA-5-methyluridine54 2-sulfurtransferase and Organism(s) Thermus thermophilus and UniProt Accession Q72LF3
for references in articles please use BRENDA:EC2.8.1.15Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
2.8.1.15 tRNA-5-methyluridine54 2-sulfurtransferaseIUBMB Comments
The enzyme, found in thermophilic bacteria and archaea, modifies the ribothymidine (5-methyluridine) residue at position 54 of tRNAs. Contains zinc and an [4Fe-4S] cluster. Some organisms, such as the archaeon Pyrococcus horikoshii, do not have a TtuB sulfur-carrier protein, and appear to use sulfide as the sulfur source.
Specify your search results
The taxonomic range for the selected organisms is: Thermus thermophilus
The expected taxonomic range for this enzyme is: Archaea, Bacteria
The expected taxonomic range for this enzyme is: Archaea, Bacteria
Reaction Schemes
+
+
+
=
+
+
+
+
[TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH
+
5-methyluracil54 in tRNA
+
=
+
+
5-methyl-2-thiouracil54 in tRNA
+
[TtuB sulfur-carrier protein]-Gly-Gly
+
+
+
=
+
+
+
+
[TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH
+
5-methyuracil54 in tRNA
+
=
+
+
5-methyl-2-thiouracil54 in tRNA
+
[TtuB sulfur-carrier protein]-Gly-Gly
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
[TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH:tRNA (5-methyluridine54-2-O)-sulfurtransferase
The enzyme, found in thermophilic bacteria and archaea, modifies the ribothymidine (5-methyluridine) residue at position 54 of tRNAs. Contains zinc and an [4Fe-4S] cluster. Some organisms, such as the archaeon Pyrococcus horikoshii, do not have a TtuB sulfur-carrier protein, and appear to use sulfide as the sulfur source.
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyluridine54 in tRNA + acceptor
AMP + diphosphate + 5-methyl-2-thiouridine54 in tRNA + [TtuB sulfur-carrier protein] + reduced acceptor
-
-
-
?
ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyuracil54 in tRNA + H2O
AMP + diphosphate + 2-thioribothymidine in tRNA + [TtuB sulfur-carrier protein]-Gly-Gly
-
-
-
?
ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyuracil54 in tRNA + H2O
AMP + diphosphate + 5-methyl-2-thiouracil54 in tRNA + [TtuB sulfur-carrier protein]-Gly-Gly
-
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyluridine54 in tRNA + acceptor
AMP + diphosphate + 5-methyl-2-thiouridine54 in tRNA + [TtuB sulfur-carrier protein] + reduced acceptor
-
-
-
?
ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyuracil54 in tRNA + H2O
AMP + diphosphate + 5-methyl-2-thiouracil54 in tRNA + [TtuB sulfur-carrier protein]-Gly-Gly
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
[4Fe-4S]-center
TtuA requires oxygen-labile [4Fe-4S]-type iron (Fe)-S clusters for its enzymatic activity. The [4Fe-4S] cluster is coordinated by three highly conserved cysteine residues, and one of the Fe atoms is exposed to the active site. The cluster is coordinated by conserved residues Cys130, Cys133, and Cys222
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Zinc
presence of zinc atoms in the protein crystal. TtuA contains five characteristic metal ion-binding motifs, CXXC/H
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50 - 70
thiolation increases 7fold when the temperature rises from 50°C to 70°C
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
physiological function
metabolism
2-Thioribothymidine (s2T) is a modified nucleoside of U, specifically found at position 54 of tRNAs from extreme thermophilic microorganisms. The function of the 2-thiocarbonyl group of s2T54 is thermostabilization of the three-dimensional structure of tRNA
metabolism
The enzyme is required for growth at temperatures above 75°C. Thiolation of thymidine54 in tRNA results in a tRNA molecule with higher thermostability
physiological function
a mutant strain without TtuA activity lacks 2-thioribothymidine in tRNA, exhibits normal growth at an optimal temperature of 7 °C, and shows growth defects at temperatures higher than 80°C
physiological function
mutation of either gene TtuA or TtuB completely abolishes thio-modification of 2-thioribothymidine, and the mutants exhibit a temperature-sensitive phenotype. TtuA is possibly an ATPase possessing a P-loop motif. TtuB is a putative thio-carrier protein. Both TtuA and TtuB are required for 2-thioribothymidine in vitro formation in the presence of cysteine and ATP
physiological function
the sulfur atom of 2-thioribothymidine in tRNA is derived from cysteine or sulfate. Specific 2-thiolation of ribothymidine, at position 54, is observed in vitro, in the presence of ATP. The 2-thiolation reaction in vitro as well as expression of 2-thiolation enzymes in vivo show strong temperature dependence
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
structure of the TtuA-TtuB complex, at a resolution of 2.5 A, showing the S transfer of TtuB to tRNA using its C-terminal thiocarboxylate group. The active site of TtuA is connected to the outside by two channels, one occupied by TtuB and the other used for tRNA binding
vapour-diffusion method at 293 K under anaerobic conditions, 2-thioribothymidinesynthetic complex TtuATtuB
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C130S
C133S
C222S
D161A
D59A
E203A
H157A
K58A
N158A
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the TtuA-TtuB complex has a strong tendency to aggregate. TtuA is unstable under aerobic conditions but stable under anaerobic conditions.
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
enzyme expression increases with increasing the temperature from 50°C to 80°C
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Chen, M.; Narai, S.; Omura, N.; Shigi, N.; Chimnaronk, S.; Tanaka, Y.; Yao, M.
Identification Crystallographic study of the 2-thioribothymidine-synthetic complex TtuA-TtuB from Thermus thermophilus
Acta Crystallogr. Sect. F
72
777.781
2016
Thermus thermophilus (Q72LF3), Thermus thermophilus DSM 7039 (Q72LF3)
Shigi, N.; Sakaguchi, Y.; Suzuki, T.; Watanabe, K.
Identification of two tRNA thiolation genes required for cell growth at extremely high temperatures
J. Biol. Chem.
281
14296-306
2006
Thermus thermophilus (Q72LF3), Thermus thermophilus DSM 7039 (Q72LF3)
Shigi, N.; Suzuki, T.; Terade, T.; Shirouzu, M.; Yokoyama, S.; Watanabe, K.
Identification Temperature-dependent biosynthesis of 2-thioribothymidine of Thermus thermophilus tRNA
J. Biol. Chem.
281
2104-2113
2006
Thermus thermophilus (Q72LF3), Thermus thermophilus DSM 7039 (Q72LF3), Thermus thermophilus HB8 / ATCC 27634 / DSM 579 (Q72LF3)
Nakagawa, H.; Kuratani, M.; Goto-Ito, S.; Ito, T.; Katsura, K.; Terada, T.; Shirouzu, M.; Sekine, S.; Shigi, N.; Yokoyama, S.
Crystallographic and mutational studies on the tRNA thiouridine synthetase TtuA
Proteins
81
1232-1244
2013
Pyrococcus horikoshii (O58038), Pyrococcus horikoshii DSM 12428 (O58038), Thermus thermophilus (Q72LF3), Thermus thermophilus DSM 7039 (Q72LF3)
Chen, M.; Narai, S.; Omura, N.; Shigi, N.; Chimnaronk, S.; Tanaka, Y.; Yao, M.
Crystallographic study of the 2-thioribothymidine-synthetic complex TtuA-TtuB from Thermus thermophilus
Acta Crystallogr. Sect. F
72
777-781
2016
Thermus thermophilus (Q72LF3), Thermus thermophilus DSM 7039 (Q72LF3)
Shigi, N.; Asai, S.; Watanabe, K.
Identification of a rhodanese-like protein involved in thiouridine biosynthesis in Thermus thermophilus tRNA
FEBS Lett.
590
4628-4637
2016
Thermus thermophilus (Q72LF3), Thermus thermophilus DSM 7039 (Q72LF3)
Shigi, N.; Sakaguchi, Y.; Suzuki, T.; Watanabe, K.
Identification of two tRNA thiolation genes required for cell growth at extremely high temperatures
J. Biol. Chem.
281
14296-14306
2006
Thermus thermophilus (Q72LF3), Thermus thermophilus DSM 7039 (Q72LF3)
Chen, M.; Asai, S.; Narai, S.; Nambu, S.; Omura, N.; Sakaguchi, Y.; Suzuki, T.; Ikeda-Saito, M.; Watanabe, K.; Yao, M.; Shigi, N.; Tanaka, Y.
Biochemical and structural characterization of oxygen-sensitive 2-thiouridine synthesis catalyzed by an iron-sulfur protein TtuA
Proc. Natl. Acad. Sci. USA
114
4954-4959
2017
Thermus thermophilus (Q72LF3), Thermus thermophilus DSM 7039 (Q72LF3)
Nakagawa, H.; Kuratani, M.; Goto-Ito, S.; Ito, T.; Katsura, K.; Terada, T.; Shirouzu, M.; Sekine, S.; Shigi, N.; Yokoyama, S.
Crystallographic and mutational studies on the tRNA thiouridine synthetase TtuA
Proteins Struct. Funct. Bioinform.
81
1232-1244
2013
Pyrococcus horikoshii (O58038), Pyrococcus horikoshii DSM 12428 (O58038), Thermus thermophilus (Q72LF3), Thermus thermophilus DSM 7039 (Q72LF3)
EXTERNAL LINKS (specific for EC-Number 2.8.1.15)
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
