TYSY_MYCTU
ID TYSY_MYCTU Reviewed; 266 AA.
AC P9WFR9; L0TD99; O33306; P67044;
DT 16-APR-2014, integrated into UniProtKB/Swiss-Prot.
DT 25-MAY-2022, sequence version 2.
DT 03-AUG-2022, entry version 39.
DE RecName: Full=Thymidylate synthase ThyA {ECO:0000303|PubMed:18493582};
DE Short=TS {ECO:0000255|HAMAP-Rule:MF_00008, ECO:0000303|PubMed:18493582};
DE Short=TSase {ECO:0000255|HAMAP-Rule:MF_00008};
DE EC=2.1.1.45 {ECO:0000255|HAMAP-Rule:MF_00008, ECO:0000269|PubMed:18493582};
GN Name=thyA {ECO:0000255|HAMAP-Rule:MF_00008}; OrderedLocusNames=Rv2764c;
GN ORFNames=MTV002.29c;
OS Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv).
OC Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae;
OC Mycobacterium; Mycobacterium tuberculosis complex.
OX NCBI_TaxID=83332;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=9634230; DOI=10.1038/31159;
RA Cole S.T., Brosch R., Parkhill J., Garnier T., Churcher C.M., Harris D.E.,
RA Gordon S.V., Eiglmeier K., Gas S., Barry C.E. III, Tekaia F., Badcock K.,
RA Basham D., Brown D., Chillingworth T., Connor R., Davies R.M., Devlin K.,
RA Feltwell T., Gentles S., Hamlin N., Holroyd S., Hornsby T., Jagels K.,
RA Krogh A., McLean J., Moule S., Murphy L.D., Oliver S., Osborne J.,
RA Quail M.A., Rajandream M.A., Rogers J., Rutter S., Seeger K., Skelton S.,
RA Squares S., Squares R., Sulston J.E., Taylor K., Whitehead S.,
RA Barrell B.G.;
RT "Deciphering the biology of Mycobacterium tuberculosis from the complete
RT genome sequence.";
RL Nature 393:537-544(1998).
RN [2]
RP PROTEIN SEQUENCE OF 2-12, AND SEQUENCE REVISION TO N-TERMINUS.
RC STRAIN=H37Rv;
RX PubMed=34915127; DOI=10.1016/j.ygeno.2021.12.001;
RA Shi J., Meng S., Wan L., Zhang Z., Jiang S., Zhu H., Dai E., Chang L.,
RA Gao H., Wan K., Zhang L., Zhao X., Liu H., Lyu Z., Zhang Y., Xu P.;
RT "Deep N-terminomics of Mycobacterium tuberculosis H37Rv extensively correct
RT annotated encoding genes.";
RL Genomics 114:292-304(2022).
RN [3]
RP DRUG RESISTANCE.
RX PubMed=15225321; DOI=10.1111/j.1365-2958.2004.04120.x;
RA Rengarajan J., Sassetti C.M., Naroditskaya V., Sloutsky A., Bloom B.R.,
RA Rubin E.J.;
RT "The folate pathway is a target for resistance to the drug para-
RT aminosalicylic acid (PAS) in mycobacteria.";
RL Mol. Microbiol. 53:275-282(2004).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY
RP REGULATION, AND SUBUNIT.
RX PubMed=18493582; DOI=10.1371/journal.pone.0002237;
RA Hunter J.H., Gujjar R., Pang C.K., Rathod P.K.;
RT "Kinetics and ligand-binding preferences of Mycobacterium tuberculosis
RT thymidylate synthases, ThyA and ThyX.";
RL PLoS ONE 3:E2237-E2237(2008).
RN [5]
RP DRUG RESISTANCE.
RX PubMed=19237648; DOI=10.1128/aac.01197-08;
RA Mathys V., Wintjens R., Lefevre P., Bertout J., Singhal A., Kiass M.,
RA Kurepina N., Wang X.M., Mathema B., Baulard A., Kreiswirth B.N., Bifani P.;
RT "Molecular genetics of para-aminosalicylic acid resistance in clinical
RT isolates and spontaneous mutants of Mycobacterium tuberculosis.";
RL Antimicrob. Agents Chemother. 53:2100-2109(2009).
RN [6]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=21969609; DOI=10.1074/mcp.m111.011627;
RA Kelkar D.S., Kumar D., Kumar P., Balakrishnan L., Muthusamy B., Yadav A.K.,
RA Shrivastava P., Marimuthu A., Anand S., Sundaram H., Kingsbury R.,
RA Harsha H.C., Nair B., Prasad T.S., Chauhan D.S., Katoch K., Katoch V.M.,
RA Kumar P., Chaerkady R., Ramachandran S., Dash D., Pandey A.;
RT "Proteogenomic analysis of Mycobacterium tuberculosis by high resolution
RT mass spectrometry.";
RL Mol. Cell. Proteomics 10:M111.011627-M111.011627(2011).
RN [7]
RP INDUCTION, DISRUPTION PHENOTYPE, AND MUTAGENESIS OF THR-205 AND VAL-264.
RC STRAIN=H37Rv;
RX PubMed=22034487; DOI=10.1099/mic.0.053983-0;
RA Fivian-Hughes A.S., Houghton J., Davis E.O.;
RT "Mycobacterium tuberculosis thymidylate synthase gene thyX is essential and
RT potentially bifunctional, while thyA deletion confers resistance to p-
RT aminosalicylic acid.";
RL Microbiology 158:308-318(2012).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) IN COMPLEXES WITH INHIBITORS;
RP 5,10-METHYLENETETRAHYDROFOLATE AND DUMP.
RA Reddy M.C.M., Bruning J.B., Harshbarger W., Sacchettini J.C.;
RT "Crystal structure of binary and ternary complexes of thymidylate synthase
RT (ThyA) from Mycobacterium tuberculosis: insights into the selectivity and
RT mode of inhibition.";
RL Submitted (JUN-2012) to the PDB data bank.
CC -!- FUNCTION: Catalyzes the reductive methylation of 2'-deoxyuridine-5'-
CC monophosphate (dUMP) to 2'-deoxythymidine-5'-monophosphate (dTMP) while
CC utilizing 5,10-methylenetetrahydrofolate (mTHF) as the methyl donor and
CC reductant in the reaction, yielding dihydrofolate (DHF) as a by-
CC product. This enzymatic reaction provides an intracellular de novo
CC source of dTMP, an essential precursor for DNA biosynthesis.
CC {ECO:0000269|PubMed:18493582}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(6R)-5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP = 7,8-
CC dihydrofolate + dTMP; Xref=Rhea:RHEA:12104, ChEBI:CHEBI:15636,
CC ChEBI:CHEBI:57451, ChEBI:CHEBI:63528, ChEBI:CHEBI:246422;
CC EC=2.1.1.45; Evidence={ECO:0000255|HAMAP-Rule:MF_00008,
CC ECO:0000269|PubMed:18493582};
CC -!- ACTIVITY REGULATION: Is potently inhibited by 5-fluoro-2'-deoxyuridine
CC 5'-monophosphate (FdUMP), and by the folate-based 1843U89.
CC {ECO:0000269|PubMed:18493582}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=4 uM for dUMP {ECO:0000269|PubMed:18493582};
CC KM=70 uM for 5,10-methylenetetrahydrofolate
CC {ECO:0000269|PubMed:18493582};
CC Note=kcat is 18 min(-1). {ECO:0000269|PubMed:18493582};
CC -!- PATHWAY: Pyrimidine metabolism; dTTP biosynthesis. {ECO:0000255|HAMAP-
CC Rule:MF_00008}.
CC -!- SUBUNIT: Homodimer. {ECO:0000255|HAMAP-Rule:MF_00008,
CC ECO:0000269|PubMed:18493582}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_00008}.
CC -!- INDUCTION: Is expressed under the exponential phase of growth, and
CC down-regulated upon reaching the stationary phase or under both
CC oxidative and nitrosative stress. Expression of thyA is significantly
CC increased within murine macrophages or under acid stress. Is expressed
CC at a higher level than thyX under all of the in vitro and in vivo
CC growth conditions tested. {ECO:0000269|PubMed:22034487}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene display an in vitro
CC growth defect, but deletion of thyA does not affect M.tuberculosis in
CC vivo growth. Moreover, thyA deletion confers PAS resistance.
CC {ECO:0000269|PubMed:22034487}.
CC -!- MISCELLANEOUS: Mutations within this gene have been associated with
CC resistance to p-aminosalicylic acid (PAS) in some PAS-resistant
CC M.tuberculosis clinical isolates and spontaneous mutants
CC (PubMed:19237648) (PubMed:15225321). It seems that functional ThyA is
CC required for M.tuberculosis to be sensitive to PAS (PubMed:22034487).
CC {ECO:0000269|PubMed:15225321, ECO:0000269|PubMed:19237648,
CC ECO:0000305|PubMed:22034487}.
CC -!- SIMILARITY: Belongs to the thymidylate synthase family. Bacterial-type
CC ThyA subfamily. {ECO:0000255|HAMAP-Rule:MF_00008}.
CC -!- SEQUENCE CAUTION:
CC Sequence=CCP45563.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000269|PubMed:34915127};
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; AL123456; CCP45563.1; ALT_INIT; Genomic_DNA.
DR PIR; C70881; C70881.
DR RefSeq; NP_217280.1; NC_000962.3.
DR RefSeq; WP_003414075.1; NC_000962.3.
DR PDB; 3QJ7; X-ray; 2.50 A; A/B/C/D=4-266.
DR PDB; 4FOA; X-ray; 2.25 A; A/B/C/D=4-266.
DR PDB; 4FOG; X-ray; 2.40 A; A/B/C/D=4-266.
DR PDB; 4FOX; X-ray; 2.30 A; A/B/C/D/E/F/G/H=4-266.
DR PDB; 4FQS; X-ray; 1.80 A; A/B=4-266.
DR PDBsum; 3QJ7; -.
DR PDBsum; 4FOA; -.
DR PDBsum; 4FOG; -.
DR PDBsum; 4FOX; -.
DR PDBsum; 4FQS; -.
DR AlphaFoldDB; P9WFR9; -.
DR SMR; P9WFR9; -.
DR STRING; 83332.Rv2764c; -.
DR BindingDB; P9WFR9; -.
DR ChEMBL; CHEMBL1795162; -.
DR PaxDb; P9WFR9; -.
DR DNASU; 887728; -.
DR GeneID; 887728; -.
DR KEGG; mtu:Rv2764c; -.
DR PATRIC; fig|83332.111.peg.3075; -.
DR TubercuList; Rv2764c; -.
DR eggNOG; COG0207; Bacteria.
DR OMA; KQYLDLC; -.
DR PhylomeDB; P9WFR9; -.
DR BioCyc; MetaCyc:G185E-7013-MON; -.
DR BRENDA; 2.1.1.45; 3445.
DR UniPathway; UPA00575; -.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0005829; C:cytosol; IBA:GO_Central.
DR GO; GO:0004799; F:thymidylate synthase activity; IDA:MTBBASE.
DR GO; GO:0006231; P:dTMP biosynthetic process; IDA:MTBBASE.
DR GO; GO:0006235; P:dTTP biosynthetic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0046079; P:dUMP catabolic process; IDA:MTBBASE.
DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW.
DR GO; GO:0046677; P:response to antibiotic; IEA:UniProtKB-KW.
DR CDD; cd00351; TS_Pyrimidine_HMase; 1.
DR Gene3D; 3.30.572.10; -; 1.
DR HAMAP; MF_00008; Thymidy_synth_bact; 1.
DR InterPro; IPR045097; Thymidate_synth/dCMP_Mease.
DR InterPro; IPR023451; Thymidate_synth/dCMP_Mease_dom.
DR InterPro; IPR036926; Thymidate_synth/dCMP_Mease_sf.
DR InterPro; IPR000398; Thymidylate_synthase.
DR InterPro; IPR020940; Thymidylate_synthase_AS.
DR PANTHER; PTHR11548; PTHR11548; 1.
DR Pfam; PF00303; Thymidylat_synt; 1.
DR PRINTS; PR00108; THYMDSNTHASE.
DR SUPFAM; SSF55831; SSF55831; 1.
DR TIGRFAMs; TIGR03284; thym_sym; 2.
DR PROSITE; PS00091; THYMIDYLATE_SYNTHASE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic resistance; Cytoplasm; Direct protein sequencing;
KW Methyltransferase; Nucleotide biosynthesis; Reference proteome;
KW Transferase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:34915127"
FT CHAIN 2..266
FT /note="Thymidylate synthase ThyA"
FT /id="PRO_0000140990"
FT ACT_SITE 149
FT /note="Nucleophile"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00008"
FT BINDING 24
FT /ligand="dUMP"
FT /ligand_id="ChEBI:CHEBI:246422"
FT /ligand_note="ligand shared between dimeric partners"
FT /note="in other chain"
FT /evidence="ECO:0000269|Ref.8, ECO:0007744|PDB:3QJ7,
FT ECO:0007744|PDB:4FOX, ECO:0007744|PDB:4FQS"
FT BINDING 54
FT /ligand="(6R)-5,10-methylene-5,6,7,8-tetrahydrofolate"
FT /ligand_id="ChEBI:CHEBI:15636"
FT /evidence="ECO:0000269|Ref.8, ECO:0007744|PDB:4FOG"
FT BINDING 129..130
FT /ligand="dUMP"
FT /ligand_id="ChEBI:CHEBI:246422"
FT /ligand_note="ligand shared between dimeric partners"
FT /evidence="ECO:0000269|Ref.8, ECO:0007744|PDB:3QJ7,
FT ECO:0007744|PDB:4FOX, ECO:0007744|PDB:4FQS"
FT BINDING 169..172
FT /ligand="dUMP"
FT /ligand_id="ChEBI:CHEBI:246422"
FT /ligand_note="ligand shared between dimeric partners"
FT /note="in other chain"
FT /evidence="ECO:0000269|Ref.8, ECO:0007744|PDB:3QJ7,
FT ECO:0007744|PDB:4FOX, ECO:0007744|PDB:4FQS"
FT BINDING 172
FT /ligand="(6R)-5,10-methylene-5,6,7,8-tetrahydrofolate"
FT /ligand_id="ChEBI:CHEBI:15636"
FT /evidence="ECO:0000269|Ref.8, ECO:0007744|PDB:4FOG"
FT BINDING 180
FT /ligand="dUMP"
FT /ligand_id="ChEBI:CHEBI:246422"
FT /ligand_note="ligand shared between dimeric partners"
FT /note="in other chain"
FT /evidence="ECO:0000269|Ref.8, ECO:0007744|PDB:3QJ7,
FT ECO:0007744|PDB:4FOX, ECO:0007744|PDB:4FQS"
FT BINDING 210..212
FT /ligand="dUMP"
FT /ligand_id="ChEBI:CHEBI:246422"
FT /ligand_note="ligand shared between dimeric partners"
FT /note="in other chain"
FT /evidence="ECO:0000269|Ref.8, ECO:0007744|PDB:3QJ7,
FT ECO:0007744|PDB:4FOX, ECO:0007744|PDB:4FQS"
FT BINDING 265
FT /ligand="(6R)-5,10-methylene-5,6,7,8-tetrahydrofolate"
FT /ligand_id="ChEBI:CHEBI:15636"
FT /evidence="ECO:0000269|Ref.8, ECO:0007744|PDB:4FOG"
FT MUTAGEN 205
FT /note="T->A: Appears to be functional by complementation
FT study."
FT /evidence="ECO:0000269|PubMed:22034487"
FT MUTAGEN 264
FT /note="V->A: Appears to be non-functional by
FT complementation study."
FT /evidence="ECO:0000269|PubMed:22034487"
FT HELIX 3..14
FT /evidence="ECO:0007829|PDB:4FQS"
FT STRAND 16..18
FT /evidence="ECO:0007829|PDB:4FQS"
FT STRAND 21..23
FT /evidence="ECO:0007829|PDB:4FOA"
FT STRAND 26..37
FT /evidence="ECO:0007829|PDB:4FQS"
FT HELIX 38..40
FT /evidence="ECO:0007829|PDB:4FQS"
FT TURN 46..49
FT /evidence="ECO:0007829|PDB:4FQS"
FT HELIX 52..64
FT /evidence="ECO:0007829|PDB:4FQS"
FT HELIX 70..74
FT /evidence="ECO:0007829|PDB:4FQS"
FT HELIX 81..83
FT /evidence="ECO:0007829|PDB:4FQS"
FT HELIX 94..100
FT /evidence="ECO:0007829|PDB:4FQS"
FT HELIX 111..121
FT /evidence="ECO:0007829|PDB:4FQS"
FT STRAND 129..131
FT /evidence="ECO:0007829|PDB:4FQS"
FT TURN 135..137
FT /evidence="ECO:0007829|PDB:4FQS"
FT HELIX 138..140
FT /evidence="ECO:0007829|PDB:4FQS"
FT STRAND 141..143
FT /evidence="ECO:0007829|PDB:4FQS"
FT STRAND 146..155
FT /evidence="ECO:0007829|PDB:4FQS"
FT STRAND 158..169
FT /evidence="ECO:0007829|PDB:4FQS"
FT TURN 170..172
FT /evidence="ECO:0007829|PDB:4FQS"
FT HELIX 173..192
FT /evidence="ECO:0007829|PDB:4FQS"
FT STRAND 195..209
FT /evidence="ECO:0007829|PDB:4FQS"
FT HELIX 210..212
FT /evidence="ECO:0007829|PDB:4FQS"
FT HELIX 213..220
FT /evidence="ECO:0007829|PDB:4FQS"
FT STRAND 229..232
FT /evidence="ECO:0007829|PDB:4FQS"
FT HELIX 238..240
FT /evidence="ECO:0007829|PDB:4FQS"
FT HELIX 243..245
FT /evidence="ECO:0007829|PDB:4FQS"
FT STRAND 246..250
FT /evidence="ECO:0007829|PDB:4FQS"
SQ SEQUENCE 266 AA; 30152 MW; 5D6DA596F096B88B CRC64;
MSIVTPYEDL LRFVLETGTP KSDRTGTGTR SLFGQQMRYD LSAGFPLLTT KKVHFKSVAY
ELLWFLRGDS NIGWLHEHGV TIWDEWASDT GELGPIYGVQ WRSWPAPSGE HIDQISAALD
LLRTDPDSRR IIVSAWNVGE IERMALPPCH AFFQFYVADG RLSCQLYQRS ADLFLGVPFN
IASYALLTHM MAAQAGLSVG EFIWTGGDCH IYDNHVEQVR LQLSREPRPY PKLLLADRDS
IFEYTYEDIV VKNYDPHPAI KAPVAV
