ID   RMLA2_ECOLI             Reviewed;         293 AA.
AC   P61887; P27831; P76755; Q2M897;
DT   07-JUN-2004, integrated into UniProtKB/Swiss-Prot.
DT   07-JUN-2004, sequence version 1.
DT   10-JUN-2026, entry version 157.
DE   RecName: Full=Glucose-1-phosphate thymidylyltransferase 2;
DE            Short=G1P-TT 2;
DE            EC=2.7.7.24 {ECO:0000269|PubMed:7559340};
DE   AltName: Full=dTDP-glucose pyrophosphorylase 2;
DE   AltName: Full=dTDP-glucose synthase 2;
GN   Name=rffH; Synonyms=rmlA2, yifG; OrderedLocusNames=b3789, JW3763;
OS   Escherichia coli (strain K12).
OC   Bacteria; Pseudomonadati; Pseudomonadota; Gammaproteobacteria;
OC   Enterobacterales; Enterobacteriaceae; Escherichia.
OX   NCBI_TaxID=83333;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RX   PubMed=1379743; DOI=10.1126/science.1379743;
RA   Daniels D.L., Plunkett G. III, Burland V.D., Blattner F.R.;
RT   "Analysis of the Escherichia coli genome: DNA sequence of the region from
RT   84.5 to 86.5 minutes.";
RL   Science 257:771-778(1992).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RX   PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA   Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA   Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA   Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA   Shao Y.;
RT   "The complete genome sequence of Escherichia coli K-12.";
RL   Science 277:1453-1462(1997).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=16738553; DOI=10.1038/msb4100049;
RA   Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA   Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT   "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT   and W3110.";
RL   Mol. Syst. Biol. 2:E1-E5(2006).
RN   [4]
RP   FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RC   STRAIN=K12;
RX   PubMed=7559340; DOI=10.1128/jb.177.19.5539-5546.1995;
RA   Marolda C.L., Valvano M.A.;
RT   "Genetic analysis of the dTDP-rhamnose biosynthesis region of the
RT   Escherichia coli VW187 (O7:K1) rfb gene cluster: identification of
RT   functional homologs of rfbB and rfbA in the rff cluster and correct
RT   location of the rffE gene.";
RL   J. Bacteriol. 177:5539-5546(1995).
RN   [5]
RP   X-RAY CRYSTALLOGRAPHY (2.6 ANGSTROMS) OF COMPLEX WITH DTTP, COFACTOR, AND
RP   SUBUNIT.
RX   PubMed=12171937; DOI=10.1074/jbc.m206932200;
RA   Sivaraman J., Sauve V., Matte A., Cygler M.;
RT   "Crystal structure of Escherichia coli glucose-1-phosphate
RT   thymidylyltransferase (RffH) complexed with dTTP and Mg2+.";
RL   J. Biol. Chem. 277:44214-44219(2002).
CC   -!- FUNCTION: Catalyzes the formation of dTDP-glucose, from dTTP and
CC       glucose 1-phosphate, as well as its pyrophosphorolysis.
CC       {ECO:0000269|PubMed:7559340}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=dTTP + alpha-D-glucose 1-phosphate + H(+) = dTDP-alpha-D-
CC         glucose + diphosphate; Xref=Rhea:RHEA:15225, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:33019, ChEBI:CHEBI:37568, ChEBI:CHEBI:57477,
CC         ChEBI:CHEBI:58601; EC=2.7.7.24;
CC         Evidence={ECO:0000269|PubMed:7559340};
CC   -!- COFACTOR:
CC       Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC         Evidence={ECO:0000269|PubMed:12171937};
CC       Note=Binds 1 Mg(2+) ion per subunit. {ECO:0000269|PubMed:12171937};
CC   -!- PATHWAY: Nucleotide-sugar biosynthesis; dTDP-4-acetamido-4,6-
CC       dideoxygalactose biosynthesis. {ECO:0000269|PubMed:7559340}.
CC   -!- PATHWAY: Bacterial outer membrane biogenesis; enterobacterial common
CC       antigen biosynthesis. {ECO:0000269|PubMed:7559340}.
CC   -!- SUBUNIT: Homotetramer; arranged as a dimer of dimers.
CC       {ECO:0000269|PubMed:12171937}.
CC   -!- SIMILARITY: Belongs to the glucose-1-phosphate thymidylyltransferase
CC       family. {ECO:0000305}.
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DR   EMBL; M87049; AAA67589.1; -; Genomic_DNA.
DR   EMBL; U00096; AAC76794.1; -; Genomic_DNA.
DR   EMBL; AP009048; BAE77509.1; -; Genomic_DNA.
DR   PIR; H65182; H65182.
DR   RefSeq; NP_418236.1; NC_000913.3.
DR   RefSeq; WP_000676056.1; NZ_CP009789.1.
DR   PDB; 1MC3; X-ray; 2.60 A; A/B=1-293.
DR   AlphaFoldDB; P61887; -.
DR   SMR; P61887; -.
DR   BioGRID; 4261364; 237.
DR   DIP; DIP-10689N; -.
DR   FunCoup; P61887; 606.
DR   IntAct; P61887; 5.
DR   STRING; 511145.b3789; -.
DR   jPOST; P61887; -.
DR   PaxDb; 511145-b3789; -.
DR   EnsemblBacteria; AAC76794; AAC76794; b3789.
DR   GeneID; 75174021; -.
DR   GeneID; 948299; -.
DR   KEGG; ecj:JW3763; -.
DR   KEGG; eco:b3789; -.
DR   KEGG; ecoc:C3026_20515; -.
DR   PATRIC; fig|1411691.4.peg.2917; -.
DR   EchoBASE; EB1423; -.
DR   eggNOG; COG1209; Bacteria.
DR   HOGENOM; CLU_029499_9_0_6; -.
DR   InParanoid; P61887; -.
DR   OMA; AWRQGWI; -.
DR   OrthoDB; 9803871at2; -.
DR   PhylomeDB; P61887; -.
DR   BioCyc; EcoCyc:DTDPGLUCOSEPP2-MONOMER; -.
DR   BioCyc; MetaCyc:DTDPGLUCOSEPP2-MONOMER; -.
DR   UniPathway; UPA00566; -.
DR   UniPathway; UPA00817; -.
DR   EvolutionaryTrace; P61887; -.
DR   PRO; PR:P61887; -.
DR   Proteomes; UP000000625; Chromosome.
DR   GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR   GO; GO:0008879; F:glucose-1-phosphate thymidylyltransferase activity; IDA:UniProtKB.
DR   GO; GO:0042802; F:identical protein binding; IDA:EcoCyc.
DR   GO; GO:0000287; F:magnesium ion binding; IDA:UniProtKB.
DR   GO; GO:0009246; P:enterobacterial common antigen biosynthetic process; IEA:UniProtKB-UniPathway.
DR   GO; GO:0009243; P:O antigen biosynthetic process; EXP:EcoCyc.
DR   GO; GO:0000271; P:polysaccharide biosynthetic process; IDA:UniProtKB.
DR   CDD; cd02538; G1P_TT_short; 1.
DR   FunFam; 3.90.550.10:FF:000023; Glucose-1-phosphate thymidylyltransferase; 1.
DR   Gene3D; 3.90.550.10; Spore Coat Polysaccharide Biosynthesis Protein SpsA, Chain A; 1.
DR   InterPro; IPR005907; G1P_thy_trans_s.
DR   InterPro; IPR005835; NTP_transferase_dom.
DR   InterPro; IPR029044; Nucleotide-diphossugar_trans.
DR   NCBIfam; TIGR01207; rmlA; 1.
DR   PANTHER; PTHR43532; GLUCOSE-1-PHOSPHATE THYMIDYLYLTRANSFERASE; 1.
DR   PANTHER; PTHR43532:SF4; GLUCOSE-1-PHOSPHATE THYMIDYLYLTRANSFERASE 2; 1.
DR   Pfam; PF00483; NTP_transferase; 1.
DR   SUPFAM; SSF53448; Nucleotide-diphospho-sugar transferases; 1.
DR   PDBsum; 1MC3; -.
PE   1: Evidence at protein level;
KW   3D-structure; Exopolysaccharide synthesis; Magnesium; Metal-binding;
KW   Nucleotidyltransferase; Reference proteome; Transferase.
FT   CHAIN           1..293
FT                   /note="Glucose-1-phosphate thymidylyltransferase 2"
FT                   /id="PRO_0000207992"
FT   BINDING         108
FT                   /ligand="Mg(2+)"
FT                   /ligand_id="ChEBI:CHEBI:18420"
FT                   /evidence="ECO:0000269|PubMed:12171937"
FT   BINDING         223
FT                   /ligand="Mg(2+)"
FT                   /ligand_id="ChEBI:CHEBI:18420"
FT                   /evidence="ECO:0000269|PubMed:12171937"
FT   CONFLICT        5
FT                   /note="I -> M (in Ref. 1; AAA67589)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        68..69
FT                   /note="GS -> VG (in Ref. 1; AAA67589)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        130..131
FT                   /note="AT -> P (in Ref. 1; AAA67589)"
FT                   /evidence="ECO:0000305"
FT   STRAND          2..6
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           12..14
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           15..18
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           23..25
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          26..28
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           35..43
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          48..53
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   TURN            55..57
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           58..65
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           69..71
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          74..79
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           88..92
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           94..97
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          102..106
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          109..112
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           117..123
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          128..136
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          141..145
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          147..149
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          152..156
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          167..175
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           179..185
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           188..190
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          191..194
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           197..206
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          210..214
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   STRAND          220..223
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           227..244
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           251..257
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           263..272
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   TURN            273..275
FT                   /evidence="ECO:0007829|PDB:1MC3"
FT   HELIX           277..284
FT                   /evidence="ECO:0007829|PDB:1MC3"
SQ   SEQUENCE   293 AA;  32734 MW;  5DA66676CD8F9139 CRC64;
     MKGIILAGGS GTRLHPITRG VSKQLLPIYD KPMIYYPLSV LMLAGIREIL IITTPEDKGY
     FQRLLGDGSE FGIQLEYAEQ PSPDGLAQAF IIGETFLNGE PSCLVLGDNI FFGQGFSPKL
     RHVAARTEGA TVFGYQVMDP ERFGVVEFDD NFRAISLEEK PKQPKSNWAV TGLYFYDSKV
     VEYAKQVKPS ERGELEITSI NQMYLEAGNL TVELLGRGFA WLDTGTHDSL IEASTFVQTV
     EKRQGFKIAC LEEIAWRNGW LDDEGVKRAA SSLAKTGYGQ YLLELLRARP RQY
//