TTCA_ECOLI
ID TTCA_ECOLI Reviewed; 311 AA.
AC P76055; Q2MBE9; Q47558;
DT 01-NOV-1997, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1997, sequence version 1.
DT 03-AUG-2022, entry version 142.
DE RecName: Full=tRNA-cytidine(32) 2-sulfurtransferase {ECO:0000255|HAMAP-Rule:MF_01850};
DE EC=2.8.1.- {ECO:0000269|PubMed:24914049};
DE AltName: Full=Two-thiocytidine biosynthesis protein A {ECO:0000303|PubMed:14729701};
DE AltName: Full=tRNA 2-thiocytidine biosynthesis protein TtcA {ECO:0000255|HAMAP-Rule:MF_01850};
GN Name=ttcA {ECO:0000255|HAMAP-Rule:MF_01850, ECO:0000303|PubMed:14729701};
GN Synonyms=ydaO; OrderedLocusNames=b1344, JW1338;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA Shao Y.;
RT "The complete genome sequence of Escherichia coli K-12.";
RL Science 277:1453-1462(1997).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=16738553; DOI=10.1038/msb4100049;
RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT and W3110.";
RL Mol. Syst. Biol. 2:E1-E5(2006).
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-40.
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=7764507; DOI=10.1271/bbb.58.117;
RA Talukder A.A., Yanai S., Yamada M.;
RT "Analysis of products of the Escherichia coli genomic genes and regulation
RT of their expressions: an applicable procedure for genomic analysis of other
RT microorganisms.";
RL Biosci. Biotechnol. Biochem. 58:117-120(1994).
RN [4]
RP FUNCTION, DISRUPTION PHENOTYPE, AND PATHWAY.
RC STRAIN=GRB105;
RX PubMed=14729701; DOI=10.1128/jb.186.3.750-757.2004;
RA Jaeger G., Leipuviene R., Pollard M.G., Qian Q., Bjoerk G.R.;
RT "The conserved Cys-X1-X2-Cys motif present in the TtcA protein is required
RT for the thiolation of cytidine in position 32 of tRNA from Salmonella
RT enterica serovar Typhimurium.";
RL J. Bacteriol. 186:750-757(2004).
RN [5]
RP CRYSTALLIZATION.
RX PubMed=16962548; DOI=10.1016/j.ab.2006.07.027;
RA Ericsson U.B., Hallberg B.M., DeTitta G.T., Dekker N., Nordlund P.;
RT "Thermofluor-based high-throughput stability optimization of proteins for
RT structural studies.";
RL Anal. Biochem. 357:289-298(2006).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, SUBUNIT, MUTAGENESIS OF CYS-45;
RP CYS-122; CYS-125; CYS-191; CYS-210 AND CYS-213, AND REACTION MECHANISM.
RX PubMed=24914049; DOI=10.1093/nar/gku508;
RA Bouvier D., Labessan N., Clemancey M., Latour J.M., Ravanat J.L.,
RA Fontecave M., Atta M.;
RT "TtcA a new tRNA-thioltransferase with an Fe-S cluster.";
RL Nucleic Acids Res. 42:7960-7970(2014).
CC -!- FUNCTION: Catalyzes the ATP-dependent 2-thiolation of cytidine in
CC position 32 of tRNA, to form 2-thiocytidine (s(2)C32). The sulfur atoms
CC are provided by the cysteine/cysteine desulfurase (IscS) system.
CC {ECO:0000255|HAMAP-Rule:MF_01850, ECO:0000269|PubMed:24914049,
CC ECO:0000305|PubMed:14729701}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=AH2 + ATP + cytidine(32) in tRNA + S-sulfanyl-L-cysteinyl-
CC [cysteine desulfurase] = 2-thiocytidine(32) in tRNA + A + AMP +
CC diphosphate + H(+) + L-cysteinyl-[cysteine desulfurase];
CC Xref=Rhea:RHEA:57048, Rhea:RHEA-COMP:10288, Rhea:RHEA-COMP:12157,
CC Rhea:RHEA-COMP:12158, Rhea:RHEA-COMP:14821, ChEBI:CHEBI:13193,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:17499, ChEBI:CHEBI:29950,
CC ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:61963,
CC ChEBI:CHEBI:82748, ChEBI:CHEBI:141453, ChEBI:CHEBI:456215;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_01850,
CC ECO:0000269|PubMed:24914049, ECO:0000305|PubMed:14729701};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:57049;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_01850,
CC ECO:0000269|PubMed:24914049, ECO:0000305|PubMed:14729701};
CC -!- COFACTOR:
CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC Evidence={ECO:0000269|PubMed:24914049};
CC Note=Binds 1 [4Fe-4S] cluster per subunit. The cluster is chelated by
CC three Cys residues, the fourth Fe has a free coordination site that may
CC bind a sulfur atom transferred from the persulfide of IscS. The [4Fe-
CC 4S] cluster is highly sensitive to oxygen and prone to decompose into
CC the [2Fe-2S] form and further degraded forms.
CC {ECO:0000269|PubMed:24914049};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000305|PubMed:24914049};
CC -!- PATHWAY: tRNA modification. {ECO:0000255|HAMAP-Rule:MF_01850,
CC ECO:0000269|PubMed:14729701}.
CC -!- SUBUNIT: Homodimer. {ECO:0000255|HAMAP-Rule:MF_01850,
CC ECO:0000269|PubMed:24914049}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_01850,
CC ECO:0000305}.
CC -!- DISRUPTION PHENOTYPE: Loss of cytidine thiolation in position 32 in
CC tRNA. {ECO:0000269|PubMed:14729701}.
CC -!- MISCELLANEOUS: The thiolation reaction likely consists of two steps: a
CC first activation step by ATP to form an adenylated intermediate of the
CC target base of tRNA, and a second nucleophilic substitution step of the
CC sulfur (S) atom supplied by the hydrosulfide attached to the Fe-S
CC cluster. {ECO:0000305|PubMed:24914049}.
CC -!- SIMILARITY: Belongs to the TtcA family. {ECO:0000255|HAMAP-
CC Rule:MF_01850}.
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DR EMBL; U00096; AAC74426.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE76407.1; -; Genomic_DNA.
DR EMBL; D21139; BAA04675.1; -; Genomic_DNA.
DR PIR; C64884; C64884.
DR RefSeq; NP_415860.1; NC_000913.3.
DR RefSeq; WP_001157406.1; NZ_CP047127.1.
DR AlphaFoldDB; P76055; -.
DR SMR; P76055; -.
DR BioGRID; 4261949; 118.
DR IntAct; P76055; 2.
DR STRING; 511145.b1344; -.
DR jPOST; P76055; -.
DR PaxDb; P76055; -.
DR PRIDE; P76055; -.
DR EnsemblBacteria; AAC74426; AAC74426; b1344.
DR EnsemblBacteria; BAE76407; BAE76407; BAE76407.
DR GeneID; 948967; -.
DR KEGG; ecj:JW1338; -.
DR KEGG; eco:b1344; -.
DR PATRIC; fig|1411691.4.peg.932; -.
DR EchoBASE; EB3140; -.
DR eggNOG; COG0037; Bacteria.
DR HOGENOM; CLU_026481_0_0_6; -.
DR InParanoid; P76055; -.
DR OMA; MNLDQKQ; -.
DR PhylomeDB; P76055; -.
DR BioCyc; EcoCyc:G6675-MON; -.
DR BioCyc; MetaCyc:G6675-MON; -.
DR PRO; PR:P76055; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IDA:EcoCyc.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-UniRule.
DR GO; GO:0000287; F:magnesium ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0016783; F:sulfurtransferase activity; IDA:EcoCyc.
DR GO; GO:0000049; F:tRNA binding; IEA:UniProtKB-KW.
DR GO; GO:0006400; P:tRNA modification; IMP:EcoCyc.
DR GO; GO:0034227; P:tRNA thio-modification; IDA:EcoCyc.
DR Gene3D; 3.40.50.620; -; 1.
DR HAMAP; MF_01850; TtcA; 1.
DR InterPro; IPR014729; Rossmann-like_a/b/a_fold.
DR InterPro; IPR011063; TilS/TtcA_N.
DR InterPro; IPR012089; tRNA_Cyd_32_2_STrfase.
DR InterPro; IPR035107; tRNA_thiolation_TtcA_Ctu1.
DR Pfam; PF01171; ATP_bind_3; 1.
DR PIRSF; PIRSF004976; ATPase_YdaO; 1.
PE 1: Evidence at protein level;
KW 4Fe-4S; ATP-binding; Cytoplasm; Iron; Iron-sulfur; Magnesium;
KW Metal-binding; Nucleotide-binding; Reference proteome; RNA-binding;
KW Transferase; tRNA processing; tRNA-binding.
FT CHAIN 1..311
FT /note="tRNA-cytidine(32) 2-sulfurtransferase"
FT /id="PRO_0000168907"
FT MOTIF 47..52
FT /note="PP-loop motif"
FT /evidence="ECO:0000305|PubMed:24914049"
FT BINDING 122
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:O58038,
FT ECO:0000305|PubMed:24914049"
FT BINDING 125
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:O58038,
FT ECO:0000305|PubMed:24914049"
FT BINDING 213
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:O58038,
FT ECO:0000305|PubMed:24914049"
FT MUTAGEN 45
FT /note="C->A: Able to functionally complement a ttcA
FT deletion mutant as efficiently as wild-type."
FT /evidence="ECO:0000269|PubMed:24914049"
FT MUTAGEN 122
FT /note="C->A: Not able to functionally complement a ttcA
FT deletion mutant."
FT /evidence="ECO:0000269|PubMed:24914049"
FT MUTAGEN 125
FT /note="C->A: Not able to functionally complement a ttcA
FT deletion mutant."
FT /evidence="ECO:0000269|PubMed:24914049"
FT MUTAGEN 191
FT /note="C->A: Able to functionally complement a ttcA
FT deletion mutant as efficiently as wild-type."
FT /evidence="ECO:0000269|PubMed:24914049"
FT MUTAGEN 210
FT /note="C->A: Able to functionally complement a ttcA
FT deletion mutant, although less efficiently than wild-type."
FT /evidence="ECO:0000269|PubMed:24914049"
FT MUTAGEN 213
FT /note="C->A: Not able to functionally complement a ttcA
FT deletion mutant."
FT /evidence="ECO:0000269|PubMed:24914049"
FT CONFLICT 11
FT /note="E -> A (in Ref. 3; BAA04675)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 311 AA; 35561 MW; D696057B7C092D79 CRC64;
MQENQQITKK EQYNLNKLQK RLRRNVGEAI ADFNMIEEGD RIMVCLSGGK DSYTMLEILR
NLQQSAPINF SLVAVNLDQK QPGFPEHVLP EYLEKLGVEY KIVEENTYGI VKEKIPEGKT
TCSLCSRLRR GILYRTATEL GATKIALGHH RDDILQTLFL NMFYGGKMKG MPPKLMSDDG
KHIVIRPLAY CREKDIQRFA DAKAFPIIPC NLCGSQPNLQ RQVIADMLRD WDKRYPGRIE
TMFSAMQNVV PSHLCDTNLF DFKGITHGSE VVNGGDLAFD REEIPLQPAC WQPEEDENQL
DELRLNVVEV K
