P32318 (THI4_YEAST) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Thiamine thiazole synthase

Alternative name(s):
Thiazole biosynthetic enzyme

Gene names

Name:	THI4
Synonyms:	ESP35, MOL1
Ordered Locus Names:	YGR144W
ORF Names:	G6620

Organism

Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast) [Reference proteome]

Taxonomic identifier

559292 [NCBI]

Taxonomic lineage

cellular organisms › Eukaryota › Opisthokonta › Fungi › Dikarya › Ascomycota › saccharomyceta › Saccharomycotina › Saccharomycetes › Saccharomycetales › Saccharomycetaceae › Saccharomyces › Saccharomyces cerevisiae

Protein attributes

Sequence length	326 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Involved in biosynthesis of the thiamine precursor thiazole. Catalyzes the conversion of NAD and glycine to adenosine diphosphate 5-(2-hydroxyethyl)-4-methylthiazole-2-carboxylic acid (ADT), an adenylated thiazole intermediate. The reaction includes an iron-dependent sulfide transfer from a conserved cysteine residue of the protein to a thiazole intermediate. The enzyme can only undergo a single turnover, which suggests it is a suicide enzyme. May have additional roles in adaptation to various stress conditions and in DNA damage tolerance. Ref.4 Ref.7 Ref.8 Ref.10 Ref.11 Ref.12 Ref.14
Cofactor	Binds 1 iron ion per subunit.
Subunit structure	Homooctamer. Ref.13
Subcellular location	Cytoplasm. Nucleus. Note: Excluded from the vacuole. Ref.4 Ref.9
Induction	Repressed by thiamine. Ref.4 Ref.6 Ref.9
Post-translational modification	During the catalytic reaction, a sulfide is transferred from Cys-205 to a reaction intermediate, generating a dehydroalanine residue. HAMAP-Rule MF_03158
Miscellaneous	Expressed at high levels in the early stationary phase of batch cultures growing on molasses, an industrial medium. HAMAP-Rule MF_03158
Sequence similarities	Belongs to the THI4 family.

Ontologies

Keywords
Biological process	Thiamine biosynthesis
Cellular component	Cytoplasm Nucleus
Ligand	Iron Metal-binding NAD
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological_process	mitochondrial genome maintenance Inferred from mutant phenotype Ref.7. Source: SGD response to stress Inferred from electronic annotation. Source: InterPro thiamine biosynthetic process Inferred from mutant phenotype Ref.6. Source: SGD thiazole biosynthetic process Inferred from direct assay Ref.14. Source: SGD
Cellular_component	cytosol Inferred from direct assay Ref.4Ref.9. Source: SGD nucleus Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	ferrous iron binding Inferred from direct assay Ref.14. Source: SGD
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
itself		3	EBI-19215,EBI-19215

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 326

326

Thiamine thiazole synthase HAMAP-Rule MF_03158

PRO_0000034058

Regions

Region

97 – 98

2

Substrate binding HAMAP-Rule MF_03158

Region

301 – 303

3

Substrate binding HAMAP-Rule MF_03158

Sites

Binding site

76

1

Substrate; via amide nitrogen

Binding site

105

1

Substrate; via amide nitrogen

Binding site

170

1

Substrate; via amide nitrogen and carbonyl oxygen

Binding site

207

1

Substrate

Binding site

237

1

Substrate

Binding site

291

1

Substrate; via amide nitrogen

Amino acid modifications

Modified residue

205

1

2,3-didehydroalanine (Cys) HAMAP-Rule MF_03158

Experimental info

Mutagenesis

97

1

E → A or Q: No activity. Ref.11

Mutagenesis

200

1

H → N: Partially active, trapping the enzyme at an advanced intermediate, just before the sulfide transfer reaction. Ref.11

Mutagenesis

204

1

C → A: Partially active, trapping the enzyme at an advanced intermediate, just before the sulfide transfer reaction. Ref.11

Mutagenesis

205

1

C → S: Partially active, with very weak activity toward NAD. Ref.11

Mutagenesis

207

1

D → A: No activity. Ref.11

Mutagenesis

237

1

H → A or N: No activity. Ref.11

Mutagenesis

238

1

D → A: Partially active, with very weak activity toward NAD. Ref.11

Mutagenesis

301

1

R → A or Q: No activity. Ref.11

Mutagenesis

304

1

P → A: No activity. Ref.11

Secondary structure

1

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326

Helix Strand Turn

Details...

Helix

17 – 19

3

Turn

25 – 29

5

Helix

45 – 62

18

Beta strand

65 – 71

7

Helix

75 – 87

13

Beta strand

93 – 96

4

Beta strand

98 – 101

4

Turn

104 – 107

4

Beta strand

116 – 119

4

Turn

120 – 122

3

Helix

123 – 128

6

Beta strand

137 – 143

7

Helix

145 – 157

13

Beta strand

162 – 165

4

Beta strand

168 – 178

11

Beta strand

180 – 183

4

Beta strand

185 – 193

9

Helix

194 – 197

4

Beta strand

209 – 214

6

Beta strand

220 – 222

3

Beta strand

230 – 233

4

Beta strand

239 – 241

3

Helix

244 – 252

9

Helix

266 – 276

11

Beta strand

286 – 288

3

Helix

291 – 297

7

Helix

307 – 325

19

Sequences

	Sequence		Length	Mass (Da)
	P32318 [UniParc]. Last modified October 1, 1993. Version 1. Checksum: 843790F2CE00BF02	FASTA	326	34,991
	10 20 30 40 50 60 MSATSTATST SASQLHLNST PVTHCLSDIV KKEDWSDFKF APIRESTVSR AMTSRYFKDL 70 80 90 100 110 120 DKFAVSDVII VGAGSSGLSA AYVIAKNRPD LKVCIIESSV APGGGSWLGG QLFSAMVMRK 130 140 150 160 170 180 PAHLFLQELE IPYEDEGDYV VVKHAALFIS TVLSKVLQLP NVKLFNATCV EDLVTRPPTE 190 200 210 220 230 240 KGEVTVAGVV TNWTLVTQAH GTQCCMDPNV IELAGYKNDG TRDLSQKHGV ILSTTGHDGP 250 260 270 280 290 300 FGAFCAKRIV DIDQNQKLGG MKGLDMNHAE HDVVIHSGAY AGVDNMYFAG MEVAELDGLN 310 320 RMGPTFGAMA LSGVHAAEQI LKHFAA

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"MOL1, a Saccharomyces cerevisiae gene that is highly expressed in early stationary phase during growth on molasses." Praekelt U.M., Meacock P.A. Yeast 8:699-710(1992) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[2]	"The nucleotide sequence of Saccharomyces cerevisiae chromosome VII." Tettelin H., Agostoni-Carbone M.L., Albermann K., Albers M., Arroyo J., Backes U., Barreiros T., Bertani I., Bjourson A.J., Brueckner M., Bruschi C.V., Carignani G., Castagnoli L., Cerdan E., Clemente M.L., Coblenz A., Coglievina M., Coissac E. , Defoor E., Del Bino S., Delius H., Delneri D., de Wergifosse P., Dujon B., Durand P., Entian K.-D., Eraso P., Escribano V., Fabiani L., Fartmann B., Feroli F., Feuermann M., Frontali L., Garcia-Gonzalez M., Garcia-Saez M.I., Goffeau A., Guerreiro P., Hani J., Hansen M., Hebling U., Hernandez K., Heumann K., Hilger F., Hofmann B., Indge K.J., James C.M., Klima R., Koetter P., Kramer B., Kramer W., Lauquin G., Leuther H., Louis E.J., Maillier E., Marconi A., Martegani E., Mazon M.J., Mazzoni C., McReynolds A.D.K., Melchioretto P., Mewes H.-W., Minenkova O., Mueller-Auer S., Nawrocki A., Netter P., Neu R., Nombela C., Oliver S.G., Panzeri L., Paoluzi S., Plevani P., Portetelle D., Portillo F., Potier S., Purnelle B., Rieger M., Riles L., Rinaldi T., Robben J., Rodrigues-Pousada C., Rodriguez-Belmonte E., Rodriguez-Torres A.M., Rose M., Ruzzi M., Saliola M., Sanchez-Perez M., Schaefer B., Schaefer M., Scharfe M., Schmidheini T., Schreer A., Skala J., Souciet J.-L., Steensma H.Y., Talla E., Thierry A., Vandenbol M., van der Aart Q.J.M., Van Dyck L., Vanoni M., Verhasselt P., Voet M., Volckaert G., Wambutt R., Watson M.D., Weber N., Wedler E., Wedler H., Wipfli P., Wolf K., Wright L.F., Zaccaria P., Zimmermann M., Zollner A., Kleine K. Nature 387:81-84(1997) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: ATCC 96604 / S288c / FY1679.
[3]	Saccharomyces Genome Database Submitted (DEC-2009) to the EMBL/GenBank/DDBJ databases Cited for: GENOME REANNOTATION. Strain: ATCC 204508 / S288c.
[4]	"Neurospora crassa CyPBP37: a cytosolic stress protein that is able to replace yeast Thi4p function in the synthesis of vitamin B1." Faou P., Tropschug M. J. Mol. Biol. 344:1147-1157(2004) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 3-10 AND 12-16, FUNCTION, SUBCELLULAR LOCATION, INDUCTION.
[5]	"The sequence of a 27 kb segment on the right arm of chromosome VII from Saccharomyces cerevisiae reveals MOL1, NAT2, RPL30B, RSR1, CYS4, PEM1/CHO2, NSR1 genes and ten new open reading frames." Skala J., Nawrocki A., Goffeau A. Yeast 11:1421-1427(1995) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 205-326. Strain: ATCC 204508 / S288c.
[6]	"Regulation of THI4 (MOL1), a thiamine-biosynthetic gene of Saccharomyces cerevisiae." Praekelt U.M., Byrne K.L., Meacock P.A. Yeast 10:481-490(1994) [PubMed] [Europe PMC] [Abstract] Cited for: THIAMINE REGULATION.
[7]	"Dual role for the yeast THI4 gene in thiamine biosynthesis and DNA damage tolerance." Machado C.R., Praekelt U.M., de Oliveira R.C., Barbosa A.C., Byrne K.L., Meacock P.A., Menck C.F. J. Mol. Biol. 273:114-121(1997) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION IN DNA DAMAGE TOLERANCE.
[8]	"Thiamin biosynthesis in eukaryotes: characterization of the enzyme-bound product of thiazole synthase from Saccharomyces cerevisiae and its implications in thiazole biosynthesis." Chatterjee A., Jurgenson C.T., Schroeder F.C., Ealick S.E., Begley T.P. J. Am. Chem. Soc. 128:7158-7159(2006) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[9]	"Pdc2 coordinates expression of the THI regulon in the yeast Saccharomyces cerevisiae." Mojzita D., Hohmann S. Mol. Genet. Genomics 276:147-161(2006) [PubMed] [Europe PMC] [Abstract] Cited for: SUBCELLULAR LOCATION, INDUCTION.
[10]	"Heat stress promotes mitochondrial instability and oxidative responses in yeast deficient in thiazole biosynthesis." Medina-Silva R., Barros M.P., Galhardo R.S., Netto L.E., Colepicolo P., Menck C.F. Res. Microbiol. 157:275-281(2006) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION IN STRESS TOLERANCE.
[11]	"Biosynthesis of thiamin thiazole in eukaryotes: conversion of NAD to an advanced intermediate." Chatterjee A., Jurgenson C.T., Schroeder F.C., Ealick S.E., Begley T.P. J. Am. Chem. Soc. 129:2914-2922(2007) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, MUTAGENESIS OF GLU-97; HIS-200; CYS-204; CYS-205; ASP-207; HIS-237; ASP-238; ARG-301 AND PRO-304.
[12]	"Biosynthesis of the thiamin-thiazole in eukaryotes: identification of a thiazole tautomer intermediate." Chatterjee A., Schroeder F.C., Jurgenson C.T., Ealick S.E., Begley T.P. J. Am. Chem. Soc. 130:11394-11398(2008) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[13]	"Structural insights into the function of the thiamin biosynthetic enzyme Thi4 from Saccharomyces cerevisiae." Jurgenson C.T., Chatterjee A., Begley T.P., Ealick S.E. Biochemistry 45:11061-11070(2006) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.82 ANGSTROMS) IN COMPLEX WITH ADT, SUBUNIT.
[14]	"Saccharomyces cerevisiae THI4p is a suicide thiamine thiazole synthase." Chatterjee A., Abeydeera N.D., Bale S., Pai P.J., Dorrestein P.C., Russell D.H., Ealick S.E., Begley T.P. Nature 478:542-546(2011) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.82 ANGSTROMS), IRON-BINDING, FUNCTION, DIDEHYDROALANINE FORMATION AT CYS-205.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

X61669 Genomic DNA. Translation: CAA43843.1.
Z72929 Genomic DNA. Translation: CAA97157.1.
Z72930 Genomic DNA. Translation: CAA97159.1.
X85807 Genomic DNA. Translation: CAA59802.1.
BK006941 Genomic DNA. Translation: DAA08235.1.

PIR

S25321.

RefSeq

NP_011660.1. NM_001181273.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
3FPZ	X-ray	1.82	A/B	1-326	[»]

ProteinModelPortal

P32318.

SMR

P32318. Positions 16-326.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-1703N.

IntAct

P32318. 5 interactions.

MINT

MINT-408091.

STRING

4932.YGR144W.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblFungi

YGR144W; YGR144W; YGR144W.

GeneID

853047.

KEGG

sce:YGR144W.

Organism-specific databases

CYGD

YGR144w.

SGD

S000003376. THI4.

Phylogenomic databases

eggNOG

COG1635.

HOGENOM

HOG000106048.

KO

K03146.

OMA

CMDPNTI.

OrthoDB

EOG45MRF9.

Enzyme and pathway databases

BioCyc

YEAST:G3O-30848-MONOMER.
YEAST:MONOMER3O-153.

Gene expression databases

Genevestigator

P32318.

GermOnline

YGR144W. Saccharomyces cerevisiae.

Family and domain databases

HAMAP

MF_03158. THI4.

InterPro

IPR027495. Sti35.
IPR002922. Thi4_fam.
[Graphical view]

PANTHER

PTHR10617:SF54. PTHR10617:SF54. 1 hit.

TIGRFAMs

TIGR00292. TIGR00292. 1 hit.

ProtoNet

Search...

Other

EvolutionaryTrace

P32318.

NextBio

972960.

Entry information

Entry name

THI4_YEAST

Accession

Primary (citable) accession number: P32318
Secondary accession number(s): D6VUS4

Entry history

Integrated into UniProtKB/Swiss-Prot:	October 1, 1993
Last sequence update:	October 1, 1993
Last modified:	May 29, 2013
This is version 105 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Fungal Protein Annotation Program