ID FOLC_MYCTU Reviewed; 487 AA.
AC I6Y0R5;
DT 15-FEB-2017, integrated into UniProtKB/Swiss-Prot.
DT 03-OCT-2012, sequence version 1.
DT 27-MAR-2024, entry version 77.
DE RecName: Full=Dihydrofolate synthase/folylpolyglutamate synthase {ECO:0000303|PubMed:23118010};
DE Short=DHFS / FPGS;
DE EC=6.3.2.12 {ECO:0000269|PubMed:23118010, ECO:0000269|PubMed:24366731};
DE EC=6.3.2.17 {ECO:0000250|UniProtKB:P08192};
DE AltName: Full=Folylpoly-gamma-glutamate synthetase;
DE AltName: Full=Tetrahydrofolylpolyglutamate synthase;
GN Name=folC {ECO:0000312|EMBL:CCP45240.1};
GN OrderedLocusNames=Rv2447c {ECO:0000312|EMBL:CCP45240.1};
OS Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv).
OC Bacteria; Actinomycetota; Actinomycetes; Mycobacteriales; 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 IDENTIFICATION, AND GENOME REANNOTATION.
RC STRAIN=ATCC 25618 / H37Rv;
RA Lew J.M.;
RL Submitted (DEC-2012) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP DISRUPTION PHENOTYPE.
RC STRAIN=H37Rv;
RX PubMed=12657046; DOI=10.1046/j.1365-2958.2003.03425.x;
RA Sassetti C.M., Boyd D.H., Rubin E.J.;
RT "Genes required for mycobacterial growth defined by high density
RT mutagenesis.";
RL Mol. Microbiol. 48:77-84(2003).
RN [4]
RP CRYSTALLIZATION, AND SUBUNIT.
RX PubMed=16754987; DOI=10.1107/s1744309106017180;
RA Young P.G., Smith C.A., Sun X., Baker E.N., Metcalf P.;
RT "Purification, crystallization and preliminary X-ray analysis of
RT Mycobacterium tuberculosis folylpolyglutamate synthase (MtbFPGS).";
RL Acta Crystallogr. F 62:579-582(2006).
RN [5]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=21969609; DOI=10.1074/mcp.m111.011445;
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 [6]
RP FUNCTION, DRUG RESISTANCE, AND PAS-RESISTANT VARIANT ALA-40.
RC STRAIN=H37Rv;
RX PubMed=23779105; DOI=10.1074/jbc.m113.475798;
RA Zheng J., Rubin E.J., Bifani P., Mathys V., Lim V., Au M., Jang J., Nam J.,
RA Dick T., Walker J.R., Pethe K., Camacho L.R.;
RT "para-Aminosalicylic acid is a prodrug targeting dihydrofolate reductase in
RT Mycobacterium tuberculosis.";
RL J. Biol. Chem. 288:23447-23456(2013).
RN [7]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RC STRAIN=H37Rv;
RX PubMed=23118010; DOI=10.1126/science.1228980;
RA Chakraborty S., Gruber T., Barry C.E. III, Boshoff H.I., Rhee K.Y.;
RT "Para-aminosalicylic acid acts as an alternative substrate of folate
RT metabolism in Mycobacterium tuberculosis.";
RL Science 339:88-91(2013).
RN [8]
RP CATALYTIC ACTIVITY, DRUG RESISTANCE, AND PAS-RESISTANT VARIANTS GLY-40;
RP ALA-43; THR-43; SER-73; GLY-150; SER-152; ALA-153 AND GLY-153.
RC STRAIN=ATCC 25177 / H37Ra;
RX PubMed=24366731; DOI=10.1128/aac.01775-13;
RA Zhao F., Wang X.D., Erber L.N., Luo M., Guo A.Z., Yang S.S., Gu J.,
RA Turman B.J., Gao Y.R., Li D.F., Cui Z.Q., Zhang Z.P., Bi L.J., Baughn A.D.,
RA Zhang X.E., Deng J.Y.;
RT "Binding pocket alterations in dihydrofolate synthase confer resistance to
RT para-aminosalicylic acid in clinical isolates of Mycobacterium
RT tuberculosis.";
RL Antimicrob. Agents Chemother. 58:1479-1487(2014).
RN [9]
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) IN COMPLEXES WITH COBALT IONS; ADP
RP AND ATP ANALOG, FUNCTION, AND DOMAIN.
RX PubMed=18566510; DOI=10.1107/s0907444908012262;
RA Young P.G., Smith C.A., Metcalf P., Baker E.N.;
RT "Structures of Mycobacterium tuberculosis folylpolyglutamate synthase
RT complexed with ADP and AMPPCP.";
RL Acta Crystallogr. D 64:745-753(2008).
CC -!- FUNCTION: Catalyzes the addition of a glutamate residue to
CC dihydropteroate (7,8-dihydropteroate or H2Pte) to form dihydrofolate
CC (7,8-dihydrofolate monoglutamate or H2Pte-Glu) (PubMed:23118010). Also
CC catalyzes successive additions of L-glutamate to tetrahydrofolate,
CC leading to folylpolyglutamate derivatives (By similarity).
CC {ECO:0000250|UniProtKB:P08192, ECO:0000269|PubMed:23118010}.
CC -!- FUNCTION: Is involved in the bioactivation of the antituberculous drug
CC para-aminosalicylic acid (PAS). Is able to use hydroxy-dihydropteroate
CC (H2PtePAS) as substrate, which is the product formed by the action of
CC DHPS (FolP1) on PAS, leading to hydroxy-dihydrofolate (H2PtePAS-Glu).
CC This compound inhibits dihydrofolate reductase DHFR (DfrA), the next
CC enzyme in the folate pathway, and thus disrupts the folate-dependent
CC metabolic pathways. {ECO:0000269|PubMed:23118010,
CC ECO:0000305|PubMed:23779105}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=7,8-dihydropteroate + ATP + L-glutamate = 7,8-dihydrofolate +
CC ADP + H(+) + phosphate; Xref=Rhea:RHEA:23584, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:17839, ChEBI:CHEBI:29985, ChEBI:CHEBI:30616,
CC ChEBI:CHEBI:43474, ChEBI:CHEBI:57451, ChEBI:CHEBI:456216;
CC EC=6.3.2.12; Evidence={ECO:0000269|PubMed:23118010,
CC ECO:0000269|PubMed:24366731};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(6S)-5,6,7,8-tetrahydrofolyl-(gamma-L-Glu)(n) + ATP + L-
CC glutamate = (6S)-5,6,7,8-tetrahydrofolyl-(gamma-L-Glu)(n+1) + ADP +
CC H(+) + phosphate; Xref=Rhea:RHEA:10580, Rhea:RHEA-COMP:14738,
CC Rhea:RHEA-COMP:14740, ChEBI:CHEBI:15378, ChEBI:CHEBI:29985,
CC ChEBI:CHEBI:30616, ChEBI:CHEBI:43474, ChEBI:CHEBI:141005,
CC ChEBI:CHEBI:456216; EC=6.3.2.17;
CC Evidence={ECO:0000250|UniProtKB:P08192};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000250|UniProtKB:P08192};
CC Note=Binds 2 Mg(2+) ions per subunit. {ECO:0000305|PubMed:18566510};
CC -!- PATHWAY: Cofactor biosynthesis; tetrahydrofolate biosynthesis; 7,8-
CC dihydrofolate from 2-amino-4-hydroxy-6-hydroxymethyl-7,8-
CC dihydropteridine diphosphate and 4-aminobenzoate: step 2/2.
CC {ECO:0000305|PubMed:23118010}.
CC -!- PATHWAY: Cofactor biosynthesis; tetrahydrofolylpolyglutamate
CC biosynthesis. {ECO:0000250|UniProtKB:P08192}.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:16754987}.
CC -!- DOMAIN: Is folded into two distinct domains, an N-terminal ATPase
CC domain and a C-terminal Rossmann-fold domain, which are joined by a
CC flexible linker. {ECO:0000269|PubMed:18566510}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene display impaired growth.
CC {ECO:0000269|PubMed:12657046}.
CC -!- MISCELLANEOUS: Mutations within this gene in the H2Pte binding pocket
CC are responsible for PAS resistance in M.tuberculosis clinical isolates
CC and laboratory strains. FolC-linked PAS resistance is mediated by
CC altered substrate specificity that results in the failure to generate
CC levels of H2PtePAS-Glu that are necessary to inhibit DHFR (DfrA);
CC therefore, a blockage of PAS bioactivation causes PAS resistance.
CC {ECO:0000269|PubMed:23779105, ECO:0000269|PubMed:24366731}.
CC -!- SIMILARITY: Belongs to the folylpolyglutamate synthase family.
CC {ECO:0000305}.
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DR EMBL; AL123456; CCP45240.1; -; Genomic_DNA.
DR RefSeq; NP_216963.1; NC_000962.3.
DR RefSeq; WP_003899324.1; NZ_NVQJ01000024.1.
DR PDB; 2VOR; X-ray; 2.30 A; A=1-487.
DR PDB; 2VOS; X-ray; 2.00 A; A=1-487.
DR PDBsum; 2VOR; -.
DR PDBsum; 2VOS; -.
DR AlphaFoldDB; I6Y0R5; -.
DR SMR; I6Y0R5; -.
DR STRING; 83332.Rv2447c; -.
DR PaxDb; 83332-Rv2447c; -.
DR DNASU; 885902; -.
DR GeneID; 885902; -.
DR KEGG; mtu:Rv2447c; -.
DR PATRIC; fig|83332.111.peg.2739; -.
DR TubercuList; Rv2447c; -.
DR eggNOG; COG0285; Bacteria.
DR HOGENOM; CLU_015869_1_2_11; -.
DR InParanoid; I6Y0R5; -.
DR OrthoDB; 9809356at2; -.
DR PhylomeDB; I6Y0R5; -.
DR BRENDA; 6.3.2.12; 3445.
DR UniPathway; UPA00077; UER00157.
DR UniPathway; UPA00850; -.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IBA:GO_Central.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0008841; F:dihydrofolate synthase activity; IBA:GO_Central.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0004326; F:tetrahydrofolylpolyglutamate synthase activity; IBA:GO_Central.
DR GO; GO:0046656; P:folic acid biosynthetic process; IEA:UniProtKB-KW.
DR GO; GO:0009396; P:folic acid-containing compound biosynthetic process; IBA:GO_Central.
DR GO; GO:0046677; P:response to antibiotic; IEA:UniProtKB-KW.
DR GO; GO:0046654; P:tetrahydrofolate biosynthetic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 3.90.190.20; Mur ligase, C-terminal domain; 1.
DR Gene3D; 3.40.1190.10; Mur-like, catalytic domain; 1.
DR InterPro; IPR001645; Folylpolyglutamate_synth.
DR InterPro; IPR018109; Folylpolyglutamate_synth_CS.
DR InterPro; IPR036565; Mur-like_cat_sf.
DR InterPro; IPR004101; Mur_ligase_C.
DR InterPro; IPR036615; Mur_ligase_C_dom_sf.
DR InterPro; IPR013221; Mur_ligase_cen.
DR NCBIfam; TIGR01499; folC; 1.
DR PANTHER; PTHR11136:SF0; DIHYDROFOLATE SYNTHETASE-RELATED; 1.
DR PANTHER; PTHR11136; FOLYLPOLYGLUTAMATE SYNTHASE-RELATED; 1.
DR Pfam; PF02875; Mur_ligase_C; 1.
DR Pfam; PF08245; Mur_ligase_M; 1.
DR SUPFAM; SSF53623; MurD-like peptide ligases, catalytic domain; 1.
DR SUPFAM; SSF53244; MurD-like peptide ligases, peptide-binding domain; 1.
DR PROSITE; PS01012; FOLYLPOLYGLU_SYNT_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic resistance; ATP-binding; Folate biosynthesis;
KW Ligase; Magnesium; Metal-binding; Nucleotide-binding; Reference proteome.
FT CHAIN 1..487
FT /note="Dihydrofolate synthase/folylpolyglutamate synthase"
FT /id="PRO_0000439040"
FT BINDING 44..46
FT /ligand="7,8-dihydropteroate"
FT /ligand_id="ChEBI:CHEBI:17839"
FT /evidence="ECO:0000250|UniProtKB:P08192"
FT BINDING 74..77
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000305|PubMed:18566510,
FT ECO:0007744|PDB:2VOR"
FT BINDING 76
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000305|PubMed:18566510"
FT BINDING 98
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000305|PubMed:18566510"
FT BINDING 150..153
FT /ligand="7,8-dihydropteroate"
FT /ligand_id="ChEBI:CHEBI:17839"
FT /evidence="ECO:0000250|UniProtKB:P08192"
FT BINDING 174
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000305|PubMed:18566510"
FT BINDING 181..183
FT /ligand="7,8-dihydropteroate"
FT /ligand_id="ChEBI:CHEBI:17839"
FT /evidence="ECO:0000250|UniProtKB:P08192"
FT BINDING 201
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000305|PubMed:18566510"
FT BINDING 203
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000305|PubMed:18566510"
FT BINDING 301
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000305|PubMed:18566510,
FT ECO:0007744|PDB:2VOR"
FT BINDING 338
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000305|PubMed:18566510,
FT ECO:0007744|PDB:2VOR"
FT BINDING 351..354
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000305|PubMed:18566510,
FT ECO:0007744|PDB:2VOR"
FT BINDING 384
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000305|PubMed:18566510"
FT VARIANT 40
FT /note="E -> A (in a spontaneous PAS-resistant mutant
FT strain; complementation with wild-type folC restores PAS
FT susceptibility, confirmimg that this mutation confers high
FT level resistance to PAS)"
FT /evidence="ECO:0000269|PubMed:23779105"
FT VARIANT 40
FT /note="E -> G (in clinical isolates: Q274 and 501063; PAS-
FT resistant; complementation with wild-type folC restores PAS
FT susceptibility)"
FT /evidence="ECO:0000269|PubMed:24366731"
FT VARIANT 43
FT /note="I -> A (in clinical isolate: Q36; PAS-resistant;
FT complementation with wild-type folC restores PAS
FT susceptibility)"
FT /evidence="ECO:0000269|PubMed:24366731"
FT VARIANT 43
FT /note="I -> T (in clinical isolates: Q449 and 1314; PAS-
FT resistant; complementation with wild-type folC restores PAS
FT susceptibility; 7-fold reduction in DHFS activity relative
FT to wild-type; loss of the ability to glutaminate H2PtePAS)"
FT /evidence="ECO:0000269|PubMed:24366731"
FT VARIANT 73
FT /note="N -> S (in a spontaneous PAS-resistant mutant
FT strain)"
FT /evidence="ECO:0000269|PubMed:24366731"
FT VARIANT 150
FT /note="S -> G (in a spontaneous PAS-resistant mutant
FT strain; complementation with wild-type folC restores PAS
FT susceptibility)"
FT /evidence="ECO:0000269|PubMed:24366731"
FT VARIANT 152
FT /note="F -> S (in a spontaneous PAS-resistant mutant
FT strain; complementation with wild-type folC restores PAS
FT susceptibility)"
FT /evidence="ECO:0000269|PubMed:24366731"
FT VARIANT 153
FT /note="E -> A (in a spontaneous PAS-resistant mutant
FT strain; complementation with wild-type folC restores PAS
FT susceptibility; 5-fold reduction in DHFS activity relative
FT to wild-type; loss of the ability to glutaminate H2PtePAS)"
FT /evidence="ECO:0000269|PubMed:24366731"
FT VARIANT 153
FT /note="E -> G (in a spontaneous PAS-resistant mutant
FT strain; complementation with wild-type folC restores PAS
FT susceptibility)"
FT /evidence="ECO:0000269|PubMed:24366731"
SQ SEQUENCE 487 AA; 50779 MW; 77D3A4E3D2DC59D0 CRC64;
MNSTNSGPPD SGSATGVVPT PDEIASLLQV EHLLDQRWPE TRIDPSLTRI SALMDLLGSP
QRSYPSIHIA GTNGKTSVAR MVDALVTALH RRTGRTTSPH LQSPVERISI DGKPISPAQY
VATYREIEPL VALIDQQSQA SAGKGGPAMS KFEVLTAMAF AAFADAPVDV AVVEVGMGGR
WDATNVINAP VAVITPISID HVDYLGADIA GIAGEKAGII TRAPDGSPDT VAVIGRQVPK
VMEVLLAESV RADASVARED SEFAVLRRQI AVGGQVLQLQ GLGGVYSDIY LPLHGEHQAH
NAVLALASVE AFFGAGAQRQ LDGDAVRAGF AAVTSPGRLE RMRSAPTVFI DAAHNPAGAS
ALAQTLAHEF DFRFLVGVLS VLGDKDVDGI LAALEPVFDS VVVTHNGSPR ALDVEALALA
AGERFGPDRV RTAENLRDAI DVATSLVDDA AADPDVAGDA FSRTGIVITG SVVTAGAART
LFGRDPQ
//
