P77718 (THII_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
tRNA sulfurtransferase
EC=2.8.1.4

Alternative name(s):
Sulfur carrier protein ThiS sulfurtransferase
Thiamine biosynthesis protein ThiI
tRNA 4-thiouridine synthase

Gene names

Name:	thiI
Synonyms:	yajK
Ordered Locus Names:	b0423, JW0413

Organism

Escherichia coli (strain K12) [Reference proteome] [HAMAP]

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia › Escherichia coli

Protein attributes

Sequence length	482 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Catalyzes the ATP-dependent transfer of a sulfur to tRNA to produce 4-thiouridine in position 8 of tRNAs, which functions as a near-UV photosensor. Also catalyzes the transfer of sulfur to the sulfur carrier protein ThiS, forming ThiS-thiocarboxylate. This is a step in the synthesis of thiazole, in the thiamine biosynthesis pathway. The sulfur is donated as persulfide by IscS. Ref.8 Ref.9 Ref.13
Catalytic activity	L-cysteine + 'activated' tRNA = L-serine + tRNA containing a thionucleotide. Ref.9 [IscS]-SSH + [ThiS]-COAMP = [IscS]-SH + [ThiS]-COSH + AMP. Ref.9
Pathway	Cofactor biosynthesis; thiamine diphosphate biosynthesis. HAMAP-Rule MF_00021
Subcellular location	Cytoplasm HAMAP-Rule MF_00021.
Sequence similarities	Belongs to the ThiI family. Contains 1 rhodanese domain. Contains 1 THUMP domain.
Biophysicochemical properties	Kinetic parameters: KM=0.24 mM for ATP Ref.13 KM=0.84 mM for GTP Vmax=1.9 nmol/min/mg enzyme with ATP as cosubstrate Vmax=1.29 nmol/min/mg enzyme with GTP as cosubstrate

Ontologies

Keywords
Biological process	Thiamine biosynthesis
Cellular component	Cytoplasm
Domain	Redox-active center
Ligand	ATP-binding Nucleotide-binding RNA-binding tRNA-binding
Molecular function	Transferase
PTM	Disulfide bond
Technical term	Complete proteome Reference proteome
Gene Ontology (GO)
Biological_process	tRNA thio-modification Inferred from electronic annotation. Source: HAMAP thiamine biosynthetic process Inferred from electronic annotation. Source: HAMAP thiamine diphosphate biosynthetic process Inferred from electronic annotation. Source: UniProtKB-UniPathway thiazole biosynthetic process Inferred from electronic annotation. Source: InterPro
Cellular_component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	ATP binding Inferred from electronic annotation. Source: HAMAP sulfurtransferase activity Inferred from electronic annotation. Source: HAMAP tRNA binding Inferred from electronic annotation. Source: HAMAP
Complete GO annotation...

Sequence annotation (Features)

	Feature key	Position(s)	Length	Description	Graphical view	Feature identifier
Molecule processing
	Chain	1 – 482	482	tRNA sulfurtransferase HAMAP-Rule MF_00021		PRO_0000154837
Regions
	Domain	61 – 165	105	THUMP
	Domain	404 – 482	79	Rhodanese
	Nucleotide binding	183 – 184	2	ATP By similarity
Sites
	Active site	456	1	Cysteine persulfide intermediate
	Binding site	265	1	ATP By similarity
	Binding site	287	1	ATP; via amide nitrogen By similarity
	Binding site	296	1	ATP By similarity
Amino acid modifications
	Disulfide bond	344 ↔ 456		Redox-active Ref.10 Ref.12
Experimental info
	Mutagenesis	108	1	C → A: No effect. Ref.10
	Mutagenesis	189	1	D → A: No activity. Ref.7
	Mutagenesis	202	1	C → A: No effect. Ref.10
	Mutagenesis	207	1	C → A: Enables partial functional complementation in vivo. Four-fold reduction in activity. Ref.10
	Mutagenesis	321	1	K → M: No activity. Ref.7
	Mutagenesis	321	1	K → R: Approximately half of wild-type activity. Ref.7
	Mutagenesis	344	1	C → A: Cannot functionally complement for wild-type enzyme in vivo. 2700-fold reduction in activity. Ref.10
	Mutagenesis	456	1	C → A: No activity. Ref.8

Sequences

	Sequence		Length	Mass (Da)
	P77718 [UniParc]. Last modified February 1, 1997. Version 1. Checksum: 72AF449459E45762	FASTA	482	54,973
	10 20 30 40 50 60 MKFIIKLFPE ITIKSQSVRL RFIKILTGNI RNVLKHYDET LAVVRHWDNI EVRAKDENQR 70 80 90 100 110 120 LAIRDALTRI PGIHHILEVE DVPFTDMHDI FEKALVQYRD QLEGKTFCVR VKRRGKHDFS 130 140 150 160 170 180 SIDVERYVGG GLNQHIESAR VKLTNPDVTV HLEVEDDRLL LIKGRYEGIG GFPIGTQEDV 190 200 210 220 230 240 LSLISGGFDS GVSSYMLMRR GCRVHYCFFN LGGAAHEIGV RQVAHYLWNR FGSSHRVRFV 250 260 270 280 290 300 AINFEPVVGE ILEKIDDGQM GVILKRMMVR AASKVAERYG VQALVTGEAL GQVSSQTLTN 310 320 330 340 350 360 LRLIDNVSDT LILRPLISYD KEHIINLARQ IGTEDFARTM PEYCGVISKS PTVKAVKSKI 370 380 390 400 410 420 EAEEEKFDFS ILDKVVEEAN NVDIREIAQQ TEQEVVEVET VNGFGPNDVI LDIRSIDEQE 430 440 450 460 470 480 DKPLKVEGID VVSLPFYKLS TKFGDLDQNK TWLLWCERGV MSRLQALYLR EQGFNNVKVY RP

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Sequence of minutes 4-25 of Escherichia coli." Chung E., Allen E., Araujo R., Aparicio A.M., Davis K., Duncan M., Federspiel N., Hyman R., Kalman S., Komp C., Kurdi O., Lew H., Lin D., Namath A., Oefner P., Roberts D., Schramm S., Davis R.W. Submitted (JAN-1997) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[2]	"The complete genome sequence of Escherichia coli K-12." Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y. Science 277:1453-1474(1997) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[3]	"Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110." Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T. Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[4]	"The biosynthesis of thiamin in Escherichia coli K-12: structural genes for thiamin biosynthetic enzymes (thiCEFGH and thiJ) and function of the thiE gene product (thiamin phosphate synthase [E.C. 2.5.1.3])." Backstrom A.D. Thesis (1996), Cornell University, United States Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 253-482. Strain: K12.
[5]	"Thiamin biosynthesis in Escherichia coli. Identification of ThiS thiocarboxylate as the immediate sulfur donor in the thiazole formation." Taylor S.V., Kelleher N.L., Kinsland C., Chiu H.-J., Costello C.A., Backstrom A.D., McLafferty F.W., Begley T.P. J. Biol. Chem. 273:16555-16560(1998) [PubMed] [Europe PMC] [Abstract] Cited for: INVOLVEMENT IN THIAMINE BIOSYNTHESIS. Strain: B/r / ATCC 12407.
[6]	"Identification of a gene involved in the generation of 4-thiouridine in tRNA." Mueller E.G., Buck C.J., Palenchar P.M., Barnhart L.E., Paulson J.L. Nucleic Acids Res. 26:2606-2610(1998) [PubMed] [Europe PMC] [Abstract] Cited for: INVOLVEMENT IN THE GENERATION OF 4-THIOURIDINE IN TRNA. Strain: RK4353.
[7]	"Using genomic information to investigate the function of ThiI, an enzyme shared between thiamin and 4-thiouridine biosynthesis." Mueller E.G., Palenchar P.M. Protein Sci. 8:2424-2427(1999) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF ASP-189 AND LYS-321.
[8]	"Evidence that ThiI, an enzyme shared between thiamin and 4-thiouridine biosynthesis, may be a sulfurtransferase that proceeds through a persulfide intermediate." Palenchar P.M., Buck C.J., Cheng H., Larson T.J., Mueller E.G. J. Biol. Chem. 275:8283-8286(2000) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, MUTAGENESIS OF CYS-456.
[9]	"Evidence for the transfer of sulfane sulfur from IscS to ThiI during the in vitro biosynthesis of 4-thiouridine in Escherichia coli tRNA." Kambampati R., Lauhon C.T. J. Biol. Chem. 275:10727-10730(2000) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION IN 4-THIOURIDINE FORMATION IN TRNA, CATALYTIC ACTIVITY, ATP DEPENDENCE.
[10]	"The role of the cysteine residues of ThiI in the generation of 4-thiouridine in tRNA." Mueller E.G., Palenchar P.M., Buck C.J. J. Biol. Chem. 276:33588-33595(2001) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF CYS-108; CYS-202; CYS-207 AND CYS-344, FORMATION OF DISULFIDE BOND DURING ENZYME CATALYSIS, REACTION MECHANISM.
[11]	"A paradigm for biological sulfur transfers via persulfide groups: a persulfide-disulfide-thiol cycle in 4-thiouridine biosynthesis." Wright C.M., Palenchar P.M., Mueller E.G. Chem. Commun. (Camb.) 22:2708-2709(2002) [PubMed] [Europe PMC] [Abstract] Cited for: REACTION MECHANISM.
[12]	"Direct evidence for enzyme persulfide and disulfide intermediates during 4-thiouridine biosynthesis." Wright C.M., Christman G.D., Snellinger A.M., Johnston M.V., Mueller E.G. Chem. Commun. (Camb.) 25:3104-3106(2006) [PubMed] [Europe PMC] [Abstract] Cited for: MASS SPECTROMETRY, CYSTEINE PERSULFIDE INTERMEDIATE, DISULFIDE BOND FORMATION, REACTION MECHANISM.
[13]	"Direct evidence that ThiI is an ATP pyrophosphatase for the adenylation of uridine in 4-thiouridine biosynthesis." You D., Xu T., Yao F., Zhou X., Deng Z. ChemBioChem 9:1879-1882(2008) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION AS PYROPHOSPHATASE, BIOPHYSICOCHEMICAL PROPERTIES, MASS SPECTROMETRY.

Cross-references

Sequence databases
EMBL GenBank DDBJ	U82664 Genomic DNA. Translation: AAB40179.1. U00096 Genomic DNA. Translation: AAC73526.1. AP009048 Genomic DNA. Translation: BAE76203.1. U34923 Genomic DNA. No translation available.
PIR	G64771.
RefSeq	NP_414957.1. NC_000913.2. YP_488715.1. NC_007779.1.
3D structure databases
ProteinModelPortal	P77718.
SMR	P77718. Positions 19-376.
ModBase	Search...
Protein-protein interaction databases
DIP	DIP-10985N.
IntAct	P77718. 12 interactions.
STRING	511145.b0423.
Proteomic databases
PaxDb	P77718.
PRIDE	P77718.
Protocols and materials databases
StructuralBiologyKnowledgebase	Search...
Genome annotation databases
EnsemblBacteria	AAC73526; AAC73526; b0423. BAE76203; BAE76203; BAE76203.
GeneID	12934350. 945067.
KEGG	ecj:Y75_p0411. eco:b0423.
PATRIC	32115997. VBIEscCol129921_0440.
Organism-specific databases
EchoBASE	EB3058.
EcoGene	EG13273. thiI.
Phylogenomic databases
eggNOG	COG0301.
HOGENOM	HOG000227469.
KO	K03151.
OMA	KLFPEIM.
ProtClustDB	PRK01269.
Enzyme and pathway databases
BioCyc	EcoCyc:THII-MONOMER. ECOL316407:JW0413-MONOMER. MetaCyc:THII-MONOMER.
UniPathway	UPA00060.
Gene expression databases
Genevestigator	P77718.
Family and domain databases
Gene3D	3.40.250.10. 1 hit. 3.40.50.620. 1 hit.
HAMAP	MF_00021. ThiI.
InterPro	IPR001763. Rhodanese-like_dom. IPR014729. Rossmann-like_a/b/a_fold. IPR026340. Thiazole_biosynth_dom. IPR003720. ThiI. IPR020536. ThiI_C_dom. IPR004114. THUMP. [Graphical view]
Pfam	PF02568. ThiI. 1 hit. PF02926. THUMP. 1 hit. [Graphical view]
SMART	SM00981. THUMP. 1 hit. [Graphical view]
SUPFAM	SSF52821. Rhodanese-like. 1 hit.
TIGRFAMs	TIGR04271. ThiI_C_thiazole. 1 hit. TIGR00342. TIGR00342. 1 hit.
PROSITE	PS50206. RHODANESE_3. 1 hit. PS51165. THUMP. 1 hit. [Graphical view]
ProtoNet	Search...

Entry information

Entry name

THII_ECOLI

Accession

Primary (citable) accession number: P77718
Secondary accession number(s): Q2MC03

Entry history

Integrated into UniProtKB/Swiss-Prot:	November 1, 1997
Last sequence update:	February 1, 1997
Last modified:	May 1, 2013
This is version 107 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program