THII_ECOLI
ID THII_ECOLI Reviewed; 482 AA.
AC P77718; Q2MC03;
DT 01-NOV-1997, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1997, sequence version 1.
DT 03-AUG-2022, entry version 163.
DE RecName: Full=tRNA sulfurtransferase;
DE EC=2.8.1.4;
DE AltName: Full=Sulfur carrier protein ThiS sulfurtransferase;
DE AltName: Full=Thiamine biosynthesis protein ThiI;
DE AltName: Full=tRNA 4-thiouridine synthase;
GN Name=thiI; Synonyms=yajK; OrderedLocusNames=b0423, JW0413;
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;
RA Chung E., Allen E., Araujo R., Aparicio A.M., Davis K., Duncan M.,
RA Federspiel N., Hyman R., Kalman S., Komp C., Kurdi O., Lew H., Lin D.,
RA Namath A., Oefner P., Roberts D., Schramm S., Davis R.W.;
RT "Sequence of minutes 4-25 of Escherichia coli.";
RL Submitted (JAN-1997) to the EMBL/GenBank/DDBJ databases.
RN [2]
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 [3]
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 [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 253-482.
RC STRAIN=K12;
RA Backstrom A.D.;
RT "The biosynthesis of thiamin in Escherichia coli K-12: structural genes for
RT thiamin biosynthetic enzymes (thiCEFGH and thiJ) and function of the thiE
RT gene product (thiamin phosphate synthase [E.C. 2.5.1.3]).";
RL Thesis (1996), Cornell University, United States.
RN [5]
RP INVOLVEMENT IN THIAMINE BIOSYNTHESIS.
RC STRAIN=B/r / ATCC 12407;
RX PubMed=9632726; DOI=10.1074/jbc.273.26.16555;
RA Taylor S.V., Kelleher N.L., Kinsland C., Chiu H.-J., Costello C.A.,
RA Backstrom A.D., McLafferty F.W., Begley T.P.;
RT "Thiamin biosynthesis in Escherichia coli. Identification of ThiS
RT thiocarboxylate as the immediate sulfur donor in the thiazole formation.";
RL J. Biol. Chem. 273:16555-16560(1998).
RN [6]
RP INVOLVEMENT IN THE GENERATION OF 4-THIOURIDINE IN TRNA.
RC STRAIN=RK4353;
RX PubMed=9592144; DOI=10.1093/nar/26.11.2606;
RA Mueller E.G., Buck C.J., Palenchar P.M., Barnhart L.E., Paulson J.L.;
RT "Identification of a gene involved in the generation of 4-thiouridine in
RT tRNA.";
RL Nucleic Acids Res. 26:2606-2610(1998).
RN [7]
RP MUTAGENESIS OF ASP-189 AND LYS-321.
RX PubMed=10595545; DOI=10.1110/ps.8.11.2424;
RA Mueller E.G., Palenchar P.M.;
RT "Using genomic information to investigate the function of ThiI, an enzyme
RT shared between thiamin and 4-thiouridine biosynthesis.";
RL Protein Sci. 8:2424-2427(1999).
RN [8]
RP FUNCTION, AND MUTAGENESIS OF CYS-456.
RX PubMed=10722656; DOI=10.1074/jbc.275.12.8283;
RA Palenchar P.M., Buck C.J., Cheng H., Larson T.J., Mueller E.G.;
RT "Evidence that ThiI, an enzyme shared between thiamin and 4-thiouridine
RT biosynthesis, may be a sulfurtransferase that proceeds through a persulfide
RT intermediate.";
RL J. Biol. Chem. 275:8283-8286(2000).
RN [9]
RP FUNCTION IN 4-THIOURIDINE FORMATION IN TRNA, CATALYTIC ACTIVITY, AND ATP
RP DEPENDENCE.
RX PubMed=10753862; DOI=10.1074/jbc.275.15.10727;
RA Kambampati R., Lauhon C.T.;
RT "Evidence for the transfer of sulfane sulfur from IscS to ThiI during the
RT in vitro biosynthesis of 4-thiouridine in Escherichia coli tRNA.";
RL J. Biol. Chem. 275:10727-10730(2000).
RN [10]
RP MUTAGENESIS OF CYS-108; CYS-202; CYS-207 AND CYS-344, FORMATION OF
RP DISULFIDE BOND DURING ENZYME CATALYSIS, AND REACTION MECHANISM.
RX PubMed=11443125; DOI=10.1074/jbc.m104067200;
RA Mueller E.G., Palenchar P.M., Buck C.J.;
RT "The role of the cysteine residues of ThiI in the generation of 4-
RT thiouridine in tRNA.";
RL J. Biol. Chem. 276:33588-33595(2001).
RN [11]
RP REACTION MECHANISM.
RX PubMed=12510310; DOI=10.1039/b208626c;
RA Wright C.M., Palenchar P.M., Mueller E.G.;
RT "A paradigm for biological sulfur transfers via persulfide groups: a
RT persulfide-disulfide-thiol cycle in 4-thiouridine biosynthesis.";
RL Chem. Commun. (Camb.) 22:2708-2709(2002).
RN [12]
RP CYSTEINE PERSULFIDE INTERMEDIATE, DISULFIDE BOND FORMATION, REACTION
RP MECHANISM, AND IDENTIFICATION BY MASS SPECTROMETRY.
RX PubMed=16855700; DOI=10.1039/b604040c;
RA Wright C.M., Christman G.D., Snellinger A.M., Johnston M.V., Mueller E.G.;
RT "Direct evidence for enzyme persulfide and disulfide intermediates during
RT 4-thiouridine biosynthesis.";
RL Chem. Commun. (Camb.) 25:3104-3106(2006).
RN [13]
RP FUNCTION AS PYROPHOSPHATASE, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP IDENTIFICATION BY MASS SPECTROMETRY.
RX PubMed=18604845; DOI=10.1002/cbic.200800033;
RA You D., Xu T., Yao F., Zhou X., Deng Z.;
RT "Direct evidence that ThiI is an ATP pyrophosphatase for the adenylation of
RT uridine in 4-thiouridine biosynthesis.";
RL ChemBioChem 9:1879-1882(2008).
CC -!- FUNCTION: Catalyzes the ATP-dependent transfer of a sulfur to tRNA to
CC produce 4-thiouridine in position 8 of tRNAs, which functions as a
CC near-UV photosensor. Also catalyzes the transfer of sulfur to the
CC sulfur carrier protein ThiS, forming ThiS-thiocarboxylate. This is a
CC step in the synthesis of thiazole, in the thiamine biosynthesis
CC pathway. The sulfur is donated as persulfide by IscS.
CC {ECO:0000269|PubMed:10722656, ECO:0000269|PubMed:10753862,
CC ECO:0000269|PubMed:18604845}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=[ThiI sulfur-carrier protein]-S-sulfanyl-L-cysteine + a
CC uridine in tRNA + ATP + H(+) + 2 reduced [2Fe-2S]-[ferredoxin] =
CC [ThiI sulfur-carrier protein]-L-cysteine + a 4-thiouridine in tRNA +
CC AMP + diphosphate + 2 oxidized [2Fe-2S]-[ferredoxin];
CC Xref=Rhea:RHEA:24176, Rhea:RHEA-COMP:10000, Rhea:RHEA-COMP:10001,
CC Rhea:RHEA-COMP:13337, Rhea:RHEA-COMP:13338, Rhea:RHEA-COMP:13339,
CC Rhea:RHEA-COMP:13340, ChEBI:CHEBI:15378, ChEBI:CHEBI:29950,
CC ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:33737,
CC ChEBI:CHEBI:33738, ChEBI:CHEBI:61963, ChEBI:CHEBI:65315,
CC ChEBI:CHEBI:136798, ChEBI:CHEBI:456215; EC=2.8.1.4;
CC Evidence={ECO:0000269|PubMed:10753862};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=[sulfur-carrier protein ThiS]-C-terminal Gly-Gly-AMP + AH2 +
CC S-sulfanyl-L-cysteinyl-[cysteine desulfurase] = [sulfur-carrier
CC protein ThiS]-C-terminal Gly-NH-CH2-C(O)SH + A + AMP + H(+) + L-
CC cysteinyl-[cysteine desulfurase]; Xref=Rhea:RHEA:43340, Rhea:RHEA-
CC COMP:12157, Rhea:RHEA-COMP:12158, Rhea:RHEA-COMP:12908, Rhea:RHEA-
CC COMP:12910, ChEBI:CHEBI:13193, ChEBI:CHEBI:15378, ChEBI:CHEBI:17499,
CC ChEBI:CHEBI:29950, ChEBI:CHEBI:61963, ChEBI:CHEBI:90618,
CC ChEBI:CHEBI:90619, ChEBI:CHEBI:456215;
CC Evidence={ECO:0000269|PubMed:10753862};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.24 mM for ATP {ECO:0000269|PubMed:18604845};
CC KM=0.84 mM for GTP {ECO:0000269|PubMed:18604845};
CC Vmax=1.9 nmol/min/mg enzyme with ATP as cosubstrate
CC {ECO:0000269|PubMed:18604845};
CC Vmax=1.29 nmol/min/mg enzyme with GTP as cosubstrate
CC {ECO:0000269|PubMed:18604845};
CC -!- PATHWAY: Cofactor biosynthesis; thiamine diphosphate biosynthesis.
CC -!- SUBCELLULAR LOCATION: Cytoplasm.
CC -!- SIMILARITY: Belongs to the ThiI family. {ECO:0000305}.
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DR EMBL; U82664; AAB40179.1; -; Genomic_DNA.
DR EMBL; U00096; AAC73526.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE76203.1; -; Genomic_DNA.
DR EMBL; U34923; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR PIR; G64771; G64771.
DR RefSeq; NP_414957.1; NC_000913.3.
DR RefSeq; WP_000668662.1; NZ_SSUW01000008.1.
DR AlphaFoldDB; P77718; -.
DR SMR; P77718; -.
DR BioGRID; 4263344; 4.
DR ComplexPortal; CPX-2140; iscS-thiI sulfurtransferase complex.
DR DIP; DIP-10985N; -.
DR IntAct; P77718; 13.
DR STRING; 511145.b0423; -.
DR jPOST; P77718; -.
DR PaxDb; P77718; -.
DR PRIDE; P77718; -.
DR EnsemblBacteria; AAC73526; AAC73526; b0423.
DR EnsemblBacteria; BAE76203; BAE76203; BAE76203.
DR GeneID; 945067; -.
DR KEGG; ecj:JW0413; -.
DR KEGG; eco:b0423; -.
DR PATRIC; fig|1411691.4.peg.1854; -.
DR EchoBASE; EB3058; -.
DR eggNOG; COG0301; Bacteria.
DR eggNOG; COG0607; Bacteria.
DR HOGENOM; CLU_037952_4_1_6; -.
DR InParanoid; P77718; -.
DR OMA; SMPEFCG; -.
DR PhylomeDB; P77718; -.
DR BioCyc; EcoCyc:THII-MON; -.
DR BioCyc; MetaCyc:THII-MON; -.
DR SABIO-RK; P77718; -.
DR UniPathway; UPA00060; -.
DR PRO; PR:P77718; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:1990221; C:L-cysteine desulfurase complex; IPI:ComplexPortal.
DR GO; GO:1990228; C:sulfurtransferase complex; IPI:ComplexPortal.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-UniRule.
DR GO; GO:0097163; F:sulfur carrier activity; IDA:EcoCyc.
DR GO; GO:0016783; F:sulfurtransferase activity; IDA:EcoCyc.
DR GO; GO:0004810; F:tRNA adenylyltransferase activity; IEA:InterPro.
DR GO; GO:0000049; F:tRNA binding; IEA:UniProtKB-UniRule.
DR GO; GO:0140741; F:tRNA U4 sulfurtransferase; IEA:UniProtKB-EC.
DR GO; GO:0009589; P:detection of UV; IC:ComplexPortal.
DR GO; GO:0019448; P:L-cysteine catabolic process; IDA:ComplexPortal.
DR GO; GO:0009228; P:thiamine biosynthetic process; IMP:EcoCyc.
DR GO; GO:0009229; P:thiamine diphosphate biosynthetic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0052837; P:thiazole biosynthetic process; IBA:GO_Central.
DR GO; GO:0002937; P:tRNA 4-thiouridine biosynthesis; IMP:EcoCyc.
DR CDD; cd01712; ThiI; 1.
DR Gene3D; 3.40.250.10; -; 1.
DR Gene3D; 3.40.50.620; -; 1.
DR HAMAP; MF_00021; ThiI; 1.
DR InterPro; IPR001763; Rhodanese-like_dom.
DR InterPro; IPR036873; Rhodanese-like_dom_sf.
DR InterPro; IPR014729; Rossmann-like_a/b/a_fold.
DR InterPro; IPR026340; Thiazole_biosynth_dom.
DR InterPro; IPR020536; ThiI_AANH.
DR InterPro; IPR004114; THUMP_dom.
DR InterPro; IPR003720; tRNA_STrfase.
DR Pfam; PF02568; ThiI; 1.
DR Pfam; PF02926; THUMP; 1.
DR SMART; SM00981; THUMP; 1.
DR SUPFAM; SSF52821; SSF52821; 1.
DR TIGRFAMs; TIGR04271; ThiI_C_thiazole; 1.
DR TIGRFAMs; TIGR00342; TIGR00342; 1.
DR PROSITE; PS50206; RHODANESE_3; 1.
DR PROSITE; PS51165; THUMP; 1.
PE 1: Evidence at protein level;
KW ATP-binding; Cytoplasm; Disulfide bond; Nucleotide-binding;
KW Redox-active center; Reference proteome; RNA-binding;
KW Thiamine biosynthesis; Transferase; tRNA-binding.
FT CHAIN 1..482
FT /note="tRNA sulfurtransferase"
FT /id="PRO_0000154837"
FT DOMAIN 61..165
FT /note="THUMP"
FT DOMAIN 404..482
FT /note="Rhodanese"
FT ACT_SITE 456
FT /note="Cysteine persulfide intermediate"
FT BINDING 183..184
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250"
FT BINDING 265
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250"
FT BINDING 287
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250"
FT BINDING 296
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250"
FT DISULFID 344..456
FT /note="Redox-active"
FT /evidence="ECO:0000269|PubMed:16855700"
FT MUTAGEN 108
FT /note="C->A: No effect."
FT /evidence="ECO:0000269|PubMed:11443125"
FT MUTAGEN 189
FT /note="D->A: No activity."
FT /evidence="ECO:0000269|PubMed:10595545"
FT MUTAGEN 202
FT /note="C->A: No effect."
FT /evidence="ECO:0000269|PubMed:11443125"
FT MUTAGEN 207
FT /note="C->A: Enables partial functional complementation in
FT vivo. Four-fold reduction in activity."
FT /evidence="ECO:0000269|PubMed:11443125"
FT MUTAGEN 321
FT /note="K->M: No activity."
FT /evidence="ECO:0000269|PubMed:10595545"
FT MUTAGEN 321
FT /note="K->R: Approximately half of wild-type activity."
FT /evidence="ECO:0000269|PubMed:10595545"
FT MUTAGEN 344
FT /note="C->A: Cannot functionally complement for wild-type
FT enzyme in vivo. 2700-fold reduction in activity."
FT /evidence="ECO:0000269|PubMed:11443125"
FT MUTAGEN 456
FT /note="C->A: No activity."
FT /evidence="ECO:0000269|PubMed:10722656"
SQ SEQUENCE 482 AA; 54973 MW; 72AF449459E45762 CRC64;
MKFIIKLFPE ITIKSQSVRL RFIKILTGNI RNVLKHYDET LAVVRHWDNI EVRAKDENQR
LAIRDALTRI PGIHHILEVE DVPFTDMHDI FEKALVQYRD QLEGKTFCVR VKRRGKHDFS
SIDVERYVGG GLNQHIESAR VKLTNPDVTV HLEVEDDRLL LIKGRYEGIG GFPIGTQEDV
LSLISGGFDS GVSSYMLMRR GCRVHYCFFN LGGAAHEIGV RQVAHYLWNR FGSSHRVRFV
AINFEPVVGE ILEKIDDGQM GVILKRMMVR AASKVAERYG VQALVTGEAL GQVSSQTLTN
LRLIDNVSDT LILRPLISYD KEHIINLARQ IGTEDFARTM PEYCGVISKS PTVKAVKSKI
EAEEEKFDFS ILDKVVEEAN NVDIREIAQQ TEQEVVEVET VNGFGPNDVI LDIRSIDEQE
DKPLKVEGID VVSLPFYKLS TKFGDLDQNK TWLLWCERGV MSRLQALYLR EQGFNNVKVY
RP
