TAGD_BACSU
ID TAGD_BACSU Reviewed; 129 AA.
AC P27623;
DT 01-AUG-1992, integrated into UniProtKB/Swiss-Prot.
DT 01-AUG-1992, sequence version 1.
DT 03-AUG-2022, entry version 168.
DE RecName: Full=Glycerol-3-phosphate cytidylyltransferase {ECO:0000303|PubMed:8393871};
DE Short=GCT;
DE Short=GCTase {ECO:0000303|PubMed:8393871};
DE Short=Gro-PCT;
DE EC=2.7.7.39 {ECO:0000269|PubMed:14506262, ECO:0000269|PubMed:8393871};
DE AltName: Full=CDP-glycerol pyrophosphorylase;
DE AltName: Full=Teichoic acid biosynthesis protein D;
GN Name=tagD {ECO:0000303|PubMed:8393871}; OrderedLocusNames=BSU35740;
OS Bacillus subtilis (strain 168).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=224308;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=168;
RX PubMed=1906926; DOI=10.1099/00221287-137-4-929;
RA Maueel C., Young M., Karamata D.;
RT "Genes concerned with synthesis of poly(glycerol phosphate), the essential
RT teichoic acid in Bacillus subtilis strain 168, are organized in two
RT divergent transcription units.";
RL J. Gen. Microbiol. 137:929-941(1991).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-10, FUNCTION,
RP CATALYTIC ACTIVITY, SUBUNIT, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE
RP SPECIFICITY, AND SUBCELLULAR LOCATION.
RC STRAIN=168 / BR151;
RX PubMed=8393871; DOI=10.1016/s0021-9258(19)85467-4;
RA Park Y.S., Sweitzer T.D., Dixon J.E., Kent C.;
RT "Expression, purification, and characterization of CTP:glycerol-3-phosphate
RT cytidylyltransferase from Bacillus subtilis.";
RL J. Biol. Chem. 268:16648-16654(1993).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=168;
RX PubMed=9384377; DOI=10.1038/36786;
RA Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V.,
RA Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R.,
RA Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S.,
RA Bruschi C.V., Caldwell B., Capuano V., Carter N.M., Choi S.-K.,
RA Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F.,
RA Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D.,
RA Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M.,
RA Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P.,
RA Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K.,
RA Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S.,
RA Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y.,
RA Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G.,
RA Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J.,
RA Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C.,
RA Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S.,
RA Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B.,
RA Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S.,
RA Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M.,
RA Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y.,
RA Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J.,
RA Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A.,
RA Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M.,
RA Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S.,
RA Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E.,
RA Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K.,
RA Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E.,
RA Yoshikawa H., Danchin A.;
RT "The complete genome sequence of the Gram-positive bacterium Bacillus
RT subtilis.";
RL Nature 390:249-256(1997).
RN [4]
RP MUTAGENESIS OF ASP-11; HIS-14; HIS-17; ASP-38; ARG-55; ARG-63; ASP-66;
RP TRP-74; HIS-84; ASP-94; ARG-113; THR-114; SER-118 AND THR-119.
RX PubMed=9182537; DOI=10.1074/jbc.272.24.15161;
RA Park Y.S., Gee P., Sanker S., Schurter E.J., Zuiderweg E.R., Kent C.;
RT "Identification of functional conserved residues of CTP:glycerol-3-
RT phosphate cytidylyltransferase. Role of histidines in the conserved HXGH in
RT catalysis.";
RL J. Biol. Chem. 272:15161-15166(1997).
RN [5]
RP REACTION MECHANISM.
RX PubMed=11487587; DOI=10.1074/jbc.m107198200;
RA Sanker S., Campbell H.A., Kent C.;
RT "Negative cooperativity of substrate binding but not enzyme activity in
RT wild-type and mutant forms of CTP:glycerol-3-phosphate
RT cytidylyltransferase.";
RL J. Biol. Chem. 276:37922-37928(2001).
RN [6]
RP REGULATION BY WALR/WALK.
RX PubMed=12950927; DOI=10.1046/j.1365-2958.2003.03661.x;
RA Howell A., Dubrac S., Andersen K.K., Noone D., Fert J., Msadek T.,
RA Devine K.;
RT "Genes controlled by the essential YycG/YycF two-component system of
RT Bacillus subtilis revealed through a novel hybrid regulator approach.";
RL Mol. Microbiol. 49:1639-1655(2003).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) OF COMPLEX WITH CTP.
RX PubMed=10508782; DOI=10.1016/s0969-2126(99)80178-6;
RA Weber C.H., Park Y.S., Sanker S., Kent C., Ludwig M.L.;
RT "A prototypical cytidylyltransferase: CTP:glycerol-3-phosphate
RT cytidylyltransferase from Bacillus subtilis.";
RL Structure 7:1113-1124(1999).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) IN COMPLEX WITH CDP-GLYCEROL,
RP CATALYTIC ACTIVITY, SUBUNIT, AND MUTAGENESIS OF LYS-44 AND LYS-46.
RX PubMed=14506262; DOI=10.1074/jbc.m306174200;
RA Pattridge K.A., Weber C.H., Friesen J.A., Sanker S., Kent C., Ludwig M.L.;
RT "Glycerol-3-phosphate cytidylyltransferase. Structural changes induced by
RT binding of CDP-glycerol and the role of lysine residues in catalysis.";
RL J. Biol. Chem. 278:51863-51871(2003).
CC -!- FUNCTION: Catalyzes the transfer of the cytidylyl group of CTP to sn-
CC glycerol 3-phosphate so the activated glycerol 3-phosphate can be used
CC for teichoic acid synthesis, via incorporation into both the linkage
CC unit and the teichoic acid polymer by TagB and TagF.
CC {ECO:0000269|PubMed:8393871}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=CTP + H(+) + sn-glycerol 3-phosphate = CDP-glycerol +
CC diphosphate; Xref=Rhea:RHEA:13361, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:37563, ChEBI:CHEBI:57597,
CC ChEBI:CHEBI:58311; EC=2.7.7.39;
CC Evidence={ECO:0000269|PubMed:14506262, ECO:0000269|PubMed:8393871};
CC -!- ACTIVITY REGULATION: Inhibited by the divalent cations cadmium,
CC mercury, tin, copper and zinc.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.85 mM for CTP {ECO:0000269|PubMed:8393871};
CC KM=3.23 mM for glycerol 3-phosphate {ECO:0000269|PubMed:8393871};
CC Vmax=185 umol/min/mg enzyme {ECO:0000269|PubMed:8393871};
CC pH dependence:
CC Optimum pH is 6.5-9.5. {ECO:0000269|PubMed:8393871};
CC Temperature dependence:
CC Optimum temperature is 50 degrees Celsius.
CC {ECO:0000269|PubMed:8393871};
CC -!- PATHWAY: Cell wall biogenesis; poly(glucopyranosyl N-
CC acetylgalactosamine 1-phosphate) teichoic acid biosynthesis.
CC -!- PATHWAY: Cell wall biogenesis; poly(glycerol phosphate) teichoic acid
CC biosynthesis.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:14506262,
CC ECO:0000269|PubMed:8393871}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000303|PubMed:8393871}.
CC -!- INDUCTION: Positively regulated by WalR. Mainly expressed during
CC exponential growth and rapidly shut off as cells enter the stationary
CC phase.
CC -!- MISCELLANEOUS: Proceeds via a random order reaction mechanism where
CC there is negative cooperativity in the binding of substrates but not in
CC catalysis.
CC -!- SIMILARITY: Belongs to the cytidylyltransferase family. {ECO:0000305}.
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DR EMBL; M57497; AAA22843.1; -; Genomic_DNA.
DR EMBL; AL009126; CAB15591.1; -; Genomic_DNA.
DR PIR; A49757; A49757.
DR RefSeq; NP_391455.1; NC_000964.3.
DR RefSeq; WP_003227921.1; NZ_JNCM01000034.1.
DR PDB; 1COZ; X-ray; 2.00 A; A/B=1-129.
DR PDB; 1N1D; X-ray; 2.00 A; A/B/C/D=1-129.
DR PDBsum; 1COZ; -.
DR PDBsum; 1N1D; -.
DR AlphaFoldDB; P27623; -.
DR SMR; P27623; -.
DR STRING; 224308.BSU35740; -.
DR DrugBank; DB02484; Cytidine 5'-diphosphoglycerol.
DR DrugBank; DB02431; Cytidine-5'-Triphosphate.
DR jPOST; P27623; -.
DR PaxDb; P27623; -.
DR PRIDE; P27623; -.
DR EnsemblBacteria; CAB15591; CAB15591; BSU_35740.
DR GeneID; 936809; -.
DR KEGG; bsu:BSU35740; -.
DR PATRIC; fig|224308.179.peg.3869; -.
DR eggNOG; COG0615; Bacteria.
DR InParanoid; P27623; -.
DR OMA; VVYFPYT; -.
DR PhylomeDB; P27623; -.
DR BioCyc; BSUB:BSU35740-MON; -.
DR BioCyc; MetaCyc:BSU35740-MON; -.
DR BRENDA; 2.7.7.39; 658.
DR UniPathway; UPA00789; -.
DR UniPathway; UPA00827; -.
DR EvolutionaryTrace; P27623; -.
DR Proteomes; UP000001570; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0047348; F:glycerol-3-phosphate cytidylyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:InterPro.
DR GO; GO:0071555; P:cell wall organization; IEA:UniProtKB-KW.
DR GO; GO:0019350; P:teichoic acid biosynthetic process; IEA:UniProtKB-KW.
DR Gene3D; 3.40.50.620; -; 1.
DR InterPro; IPR004821; Cyt_trans-like.
DR InterPro; IPR006409; G3P_cytidylTrfase.
DR InterPro; IPR014729; Rossmann-like_a/b/a_fold.
DR Pfam; PF01467; CTP_transf_like; 1.
DR TIGRFAMs; TIGR00125; cyt_tran_rel; 1.
DR TIGRFAMs; TIGR01518; g3p_cytidyltrns; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cell wall biogenesis/degradation; Cytoplasm;
KW Direct protein sequencing; Nucleotidyltransferase; Reference proteome;
KW Teichoic acid biosynthesis; Transferase.
FT CHAIN 1..129
FT /note="Glycerol-3-phosphate cytidylyltransferase"
FT /id="PRO_0000208466"
FT BINDING 9..10
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /evidence="ECO:0000269|PubMed:10508782,
FT ECO:0000269|PubMed:14506262"
FT BINDING 14..17
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /evidence="ECO:0000269|PubMed:10508782,
FT ECO:0000269|PubMed:14506262"
FT BINDING 44
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:14506262"
FT BINDING 46
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /evidence="ECO:0000269|PubMed:10508782,
FT ECO:0000269|PubMed:14506262"
FT BINDING 77
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:14506262"
FT BINDING 113..120
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /evidence="ECO:0000269|PubMed:10508782,
FT ECO:0000269|PubMed:14506262"
FT MUTAGEN 11
FT /note="D->A,E: 0.1% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 14
FT /note="H->A: Complete loss of activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 17
FT /note="H->A: Complete loss of activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 38
FT /note="D->A: 8% of wild-type activity, but 7-fold decrease
FT in substrate affinity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 44
FT /note="K->A: Reduces affinity for glycerol 3-phosphate
FT about 100-fold."
FT /evidence="ECO:0000269|PubMed:14506262"
FT MUTAGEN 46
FT /note="K->A: Reduces affinity for glycerol 3-phosphate
FT about 100-fold."
FT /evidence="ECO:0000269|PubMed:14506262"
FT MUTAGEN 55
FT /note="R->A: 0.25% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 55
FT /note="R->K: 23% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 63
FT /note="R->A: 14% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 66
FT /note="D->A: Complete loss of activity, and widespread
FT change in 3D-structure."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 66
FT /note="D->E: 16% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 74
FT /note="W->A: 50% of wild-type activity, but 9-fold decrease
FT in substrate affinity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 84
FT /note="H->A: Complete loss of activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 94
FT /note="D->A: 18% of wild-type activity, but 100-fold
FT decrease in substrate affinity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 113
FT /note="R->A: 1.75% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 113
FT /note="R->K: Complete loss of activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 114
FT /note="T->A: 9% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 118
FT /note="S->A: 6% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT MUTAGEN 119
FT /note="T->A: 8% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9182537"
FT STRAND 3..8
FT /evidence="ECO:0007829|PDB:1COZ"
FT HELIX 15..25
FT /evidence="ECO:0007829|PDB:1COZ"
FT STRAND 28..36
FT /evidence="ECO:0007829|PDB:1COZ"
FT HELIX 38..44
FT /evidence="ECO:0007829|PDB:1COZ"
FT HELIX 52..59
FT /evidence="ECO:0007829|PDB:1COZ"
FT STRAND 67..71
FT /evidence="ECO:0007829|PDB:1COZ"
FT HELIX 74..76
FT /evidence="ECO:0007829|PDB:1N1D"
FT HELIX 77..83
FT /evidence="ECO:0007829|PDB:1COZ"
FT STRAND 87..92
FT /evidence="ECO:0007829|PDB:1COZ"
FT HELIX 93..95
FT /evidence="ECO:0007829|PDB:1COZ"
FT TURN 96..99
FT /evidence="ECO:0007829|PDB:1COZ"
FT HELIX 100..102
FT /evidence="ECO:0007829|PDB:1COZ"
FT TURN 103..105
FT /evidence="ECO:0007829|PDB:1COZ"
FT STRAND 106..111
FT /evidence="ECO:0007829|PDB:1COZ"
FT HELIX 119..124
FT /evidence="ECO:0007829|PDB:1COZ"
SQ SEQUENCE 129 AA; 15272 MW; B07F532D9AAE0869 CRC64;
MKKVITYGTF DLLHWGHIKL LERAKQLGDY LVVAISTDEF NLQKQKKAYH SYEHRKLILE
TIRYVDEVIP EKNWEQKKQD IIDHNIDVFV MGDDWEGKFD FLKDQCEVVY LPRTEGISTT
KIKEEIAGL
