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 27-MAR-2024, entry version 175. 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; Bacillota; 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}. 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; 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; 224308-BSU35740; -. DR EnsemblBacteria; CAB15591; CAB15591; BSU_35740. DR GeneID; 83887462; -. DR GeneID; 936809; -. DR KEGG; bsu:BSU35740; -. DR PATRIC; fig|224308.179.peg.3869; -. DR eggNOG; COG0615; Bacteria. DR InParanoid; P27623; -. DR OrthoDB; 9802794at2; -. DR PhylomeDB; P27623; -. DR BioCyc; BSUB:BSU35740-MONOMER; -. DR BioCyc; MetaCyc:BSU35740-MONOMER; -. 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 CDD; cd02171; G3P_Cytidylyltransferase; 1. DR Gene3D; 3.40.50.620; HUPs; 1. DR InterPro; IPR004821; Cyt_trans-like. DR InterPro; IPR006409; G3P_cytidylTrfase. DR InterPro; IPR014729; Rossmann-like_a/b/a_fold. DR NCBIfam; TIGR00125; cyt_tran_rel; 1. DR NCBIfam; TIGR01518; g3p_cytidyltrns; 1. DR PANTHER; PTHR43793; FAD SYNTHASE; 1. DR PANTHER; PTHR43793:SF1; FAD SYNTHASE; 1. DR Pfam; PF01467; CTP_transf_like; 1. DR SUPFAM; SSF52374; Nucleotidylyl transferase; 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 //