ID OTSA_ECOLI Reviewed; 474 AA. AC P31677; DT 01-JUL-1993, integrated into UniProtKB/Swiss-Prot. DT 23-JAN-2007, sequence version 3. DT 27-MAR-2024, entry version 172. DE RecName: Full=Trehalose-6-phosphate synthase {ECO:0000303|PubMed:3131312}; DE Short=TPS {ECO:0000303|PubMed:3131312}; DE EC=2.4.1.15 {ECO:0000305|PubMed:20077550, ECO:0000305|PubMed:3131312}; DE AltName: Full=Alpha,alpha-trehalose-phosphate synthase [UDP-forming] {ECO:0000305}; DE AltName: Full=Osmoregulatory trehalose synthesis protein A {ECO:0000303|PubMed:3131312}; DE Short=OtsA {ECO:0000303|PubMed:3131312}; DE AltName: Full=UDP-glucose-glucosephosphate glucosyltransferase {ECO:0000305}; GN Name=otsA {ECO:0000303|PubMed:3131312}; GN OrderedLocusNames=b1896, JW5312; OS Escherichia coli (strain K12). OC Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales; OC Enterobacteriaceae; Escherichia. OX NCBI_TaxID=83333; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PARTIAL PROTEIN SEQUENCE. RC STRAIN=K12 / CSH7; RX PubMed=8045430; DOI=10.1016/0378-1119(94)90316-6; RA Kaasen I., McDougall J., Stroem A.R.; RT "Analysis of the otsBA operon for osmoregulatory trehalose synthesis in RT Escherichia coli and homology of the OtsA and OtsB proteins to the yeast RT trehalose-6-phosphate synthase/phosphatase complex."; RL Gene 145:9-15(1994). RN [2] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911; RX PubMed=9097040; DOI=10.1093/dnares/3.6.379; RA Itoh T., Aiba H., Baba T., Fujita K., Hayashi K., Inada T., Isono K., RA Kasai H., Kimura S., Kitakawa M., Kitagawa M., Makino K., Miki T., RA Mizobuchi K., Mori H., Mori T., Motomura K., Nakade S., Nakamura Y., RA Nashimoto H., Nishio Y., Oshima T., Saito N., Sampei G., Seki Y., RA Sivasundaram S., Tagami H., Takeda J., Takemoto K., Wada C., Yamamoto Y., RA Horiuchi T.; RT "A 460-kb DNA sequence of the Escherichia coli K-12 genome corresponding to RT the 40.1-50.0 min region on the linkage map."; RL DNA Res. 3:379-392(1996). RN [3] 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 [4] 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 [5] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 430-474. RC STRAIN=K12 / EMG2; RA Estep P., O'Keeffe T., Robison K., Church G.M.; RL Submitted (MAY-1995) to the EMBL/GenBank/DDBJ databases. RN [6] RP FUNCTION AS A TREHALOSE-6-PHOSPHATE SYNTHASE, CATALYTIC ACTIVITY, ACTIVITY RP REGULATION, DISRUPTION PHENOTYPE, INDUCTION, NOMENCLATURE, AND PATHWAY. RX PubMed=3131312; DOI=10.1128/jb.170.6.2841-2849.1988; RA Giaever H.M., Styrvold O.B., Kaasen I., Stroem A.R.; RT "Biochemical and genetic characterization of osmoregulatory trehalose RT synthesis in Escherichia coli."; RL J. Bacteriol. 170:2841-2849(1988). RN [7] RP INDUCTION. RX PubMed=1744047; DOI=10.1128/jb.173.24.7918-7924.1991; RA Hengge-Aronis R., Klein W., Lange R., Rimmele M., Boos W.; RT "Trehalose synthesis genes are controlled by the putative sigma factor RT encoded by rpoS and are involved in stationary-phase thermotolerance in RT Escherichia coli."; RL J. Bacteriol. 173:7918-7924(1991). RN [8] RP FUNCTION AS A TREHALOSE-6-PHOSPHATE SYNTHASE, AND INDUCTION. RX PubMed=1310094; DOI=10.1128/jb.174.3.889-898.1992; RA Kaasen I., Falkenberg P., Styrvold O.B., Strom A.R.; RT "Molecular cloning and physical mapping of the otsBA genes, which encode RT the osmoregulatory trehalose pathway of Escherichia coli: evidence that RT transcription is activated by katF (AppR)."; RL J. Bacteriol. 174:889-898(1992). RN [9] RP FUNCTION AS A TREHALOSE-6-PHOSPHATE SYNTHASE, AND INDUCTION. RC STRAIN=K12 / MC4100 / ATCC 35695 / DSM 6574; RX PubMed=12105274; DOI=10.1073/pnas.142314099; RA Kandror O., DeLeon A., Goldberg A.L.; RT "Trehalose synthesis is induced upon exposure of Escherichia coli to cold RT and is essential for viability at low temperatures."; RL Proc. Natl. Acad. Sci. U.S.A. 99:9727-9732(2002). RN [10] RP X-RAY CRYSTALLOGRAPHY (2.43 ANGSTROMS) OF 1-457 IN COMPLEX WITH SUBSTRATE RP ANALOG AND GLUCOSE-6-PHOSPHATE, AND SUBUNIT. RX PubMed=12498887; DOI=10.1016/s1074-5521(02)00292-2; RA Gibson R.P., Turkenburg J.P., Charnock S.J., Lloyd R., Davies G.J.; RT "Insights into trehalose synthesis provided by the structure of the RT retaining glucosyltransferase OtsA."; RL Chem. Biol. 9:1337-1346(2002). RN [11] RP X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOG. RX PubMed=14570926; DOI=10.1074/jbc.m307643200; RA Gibson R.P., Tarling C.A., Roberts S., Withers S.G., Davies G.J.; RT "The donor subsite of trehalose-6-phosphate synthase: binary complexes with RT UDP-glucose and UDP-2-deoxy-2-fluoro-glucose at 2 A resolution."; RL J. Biol. Chem. 279:1950-1955(2004). RN [12] RP X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOGS, RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY RP REGULATION, AND REACTION MECHANISM. RX PubMed=20077550; DOI=10.1002/anie.200905096; RA Errey J.C., Lee S.S., Gibson R.P., Martinez Fleites C., Barry C.S., RA Jung P.M., O'Sullivan A.C., Davis B.G., Davies G.J.; RT "Mechanistic insight into enzymatic glycosyl transfer with retention of RT configuration through analysis of glycomimetic inhibitors."; RL Angew. Chem. Int. Ed. Engl. 49:1234-1237(2010). CC -!- FUNCTION: Catalyzes the transfer of glucose from UDP-alpha-D-glucose CC (UDP-Glc) to D-glucose 6-phosphate (Glc-6-P) to form trehalose-6- CC phosphate. Acts with retention of the anomeric configuration of the CC UDP-sugar donor. Essential for viability of the cells at low CC temperatures and at elevated osmotic strength. CC {ECO:0000269|PubMed:12105274, ECO:0000269|PubMed:1310094, CC ECO:0000269|PubMed:20077550, ECO:0000269|PubMed:3131312}. CC -!- CATALYTIC ACTIVITY: CC Reaction=D-glucose 6-phosphate + UDP-alpha-D-glucose = alpha,alpha- CC trehalose 6-phosphate + H(+) + UDP; Xref=Rhea:RHEA:18889, CC ChEBI:CHEBI:15378, ChEBI:CHEBI:58223, ChEBI:CHEBI:58429, CC ChEBI:CHEBI:58885, ChEBI:CHEBI:61548; EC=2.4.1.15; CC Evidence={ECO:0000305|PubMed:20077550, ECO:0000305|PubMed:3131312}; CC -!- ACTIVITY REGULATION: Activated by potassium and other monovalent CC cations at 0.25 M, but partially inhibited at greater concentrations CC (PubMed:3131312). Inhibited by validoxylamine A 6'-O-phosphate CC (PubMed:20077550). {ECO:0000269|PubMed:20077550, CC ECO:0000269|PubMed:3131312}. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=1.7 mM for UDP-Glc {ECO:0000269|PubMed:20077550}; CC KM=7.3 mM for Glc-6-P {ECO:0000269|PubMed:20077550}; CC Note=kcat is 34 sec(-1) for glucosyltransferase activity. CC {ECO:0000269|PubMed:20077550}; CC -!- PATHWAY: Glycan biosynthesis; trehalose biosynthesis. CC {ECO:0000305|PubMed:3131312}. CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:12498887}. CC -!- INDUCTION: By cold-shock, osmotic-shock and during the transition to CC stationary phase. Expression is partially dependent on RpoS. CC {ECO:0000269|PubMed:12105274, ECO:0000269|PubMed:1310094, CC ECO:0000269|PubMed:1744047, ECO:0000269|PubMed:3131312}. CC -!- DISRUPTION PHENOTYPE: Mutants are viable, but osmotically sensitive in CC minimal media and sensitive to cold shock. CC {ECO:0000269|PubMed:3131312}. CC -!- SIMILARITY: Belongs to the glycosyltransferase 20 family. CC {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; X69160; CAA48913.1; -; Genomic_DNA. DR EMBL; U00096; AAC74966.1; -; Genomic_DNA. DR EMBL; AP009048; BAA15717.2; -; Genomic_DNA. DR EMBL; U27211; AAA68604.1; -; Genomic_DNA. DR PIR; I83402; I83402. DR RefSeq; NP_416410.1; NC_000913.3. DR RefSeq; WP_001295646.1; NZ_STEB01000026.1. DR PDB; 1GZ5; X-ray; 2.43 A; A/B/C/D=2-457. DR PDB; 1UQT; X-ray; 2.00 A; A/B=2-474. DR PDB; 1UQU; X-ray; 2.00 A; A/B=2-474. DR PDB; 2WTX; X-ray; 2.20 A; A/B/C/D=1-474. DR PDB; 6JAK; X-ray; 2.41 A; A/B/C/D=2-456. DR PDBsum; 1GZ5; -. DR PDBsum; 1UQT; -. DR PDBsum; 1UQU; -. DR PDBsum; 2WTX; -. DR PDBsum; 6JAK; -. DR AlphaFoldDB; P31677; -. DR SMR; P31677; -. DR BioGRID; 4259473; 16. DR DIP; DIP-10417N; -. DR IntAct; P31677; 10. DR STRING; 511145.b1896; -. DR DrugBank; DB02007; alpha-D-glucose 6-phosphate. DR DrugBank; DB03366; Imidazole. DR DrugBank; DB01861; Uridine diphosphate glucose. DR DrugBank; DB03435; Uridine-5'-Diphosphate. DR DrugBank; DB03488; Uridine-5'-diphosphate-2-deoxy-2-fluoro-alpha-D-galactose. DR CAZy; GT20; Glycosyltransferase Family 20. DR jPOST; P31677; -. DR PaxDb; 511145-b1896; -. DR EnsemblBacteria; AAC74966; AAC74966; b1896. DR GeneID; 75202700; -. DR GeneID; 946405; -. DR KEGG; ecj:JW5312; -. DR KEGG; eco:b1896; -. DR PATRIC; fig|1411691.4.peg.353; -. DR EchoBASE; EB1701; -. DR eggNOG; COG0380; Bacteria. DR HOGENOM; CLU_002351_7_1_6; -. DR InParanoid; P31677; -. DR OMA; RTIWPLF; -. DR OrthoDB; 9815690at2; -. DR PhylomeDB; P31677; -. DR BioCyc; EcoCyc:TREHALOSE6PSYN-MONOMER; -. DR BioCyc; MetaCyc:TREHALOSE6PSYN-MONOMER; -. DR BRENDA; 2.4.1.15; 2026. DR BRENDA; 3.1.3.12; 2026. DR UniPathway; UPA00299; -. DR EvolutionaryTrace; P31677; -. DR PRO; PR:P31677; -. DR Proteomes; UP000000318; Chromosome. DR Proteomes; UP000000625; Chromosome. DR GO; GO:0003825; F:alpha,alpha-trehalose-phosphate synthase (UDP-forming) activity; IDA:EcoCyc. DR GO; GO:0006974; P:DNA damage response; IEP:EcoliWiki. DR GO; GO:0006970; P:response to osmotic stress; IMP:EcoCyc. DR GO; GO:0005992; P:trehalose biosynthetic process; IMP:EcoCyc. DR GO; GO:0070415; P:trehalose metabolism in response to cold stress; IMP:EcoCyc. DR CDD; cd03788; GT20_TPS; 1. DR Gene3D; 3.40.50.2000; Glycogen Phosphorylase B; 2. DR InterPro; IPR001830; Glyco_trans_20. DR InterPro; IPR012766; Trehalose_OtsA. DR NCBIfam; TIGR02400; trehalose_OtsA; 1. DR PANTHER; PTHR10788:SF106; BCDNA.GH08860; 1. DR PANTHER; PTHR10788; TREHALOSE-6-PHOSPHATE SYNTHASE; 1. DR Pfam; PF00982; Glyco_transf_20; 1. DR SUPFAM; SSF53756; UDP-Glycosyltransferase/glycogen phosphorylase; 1. PE 1: Evidence at protein level; KW 3D-structure; Direct protein sequencing; Glycosyltransferase; Potassium; KW Reference proteome; Stress response; Transferase. FT INIT_MET 1 FT /note="Removed" FT CHAIN 2..474 FT /note="Trehalose-6-phosphate synthase" FT /id="PRO_0000122489" FT BINDING 10 FT /ligand="D-glucose 6-phosphate" FT /ligand_id="ChEBI:CHEBI:61548" FT /evidence="ECO:0000269|PubMed:12498887, FT ECO:0000269|PubMed:20077550, ECO:0007744|PDB:1GZ5, FT ECO:0007744|PDB:2WTX" FT BINDING 22..23 FT /ligand="UDP-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:58885" FT /evidence="ECO:0000305|PubMed:12498887, FT ECO:0000305|PubMed:14570926, ECO:0000305|PubMed:20077550, FT ECO:0007744|PDB:1GZ5, ECO:0007744|PDB:2WTX" FT BINDING 77 FT /ligand="D-glucose 6-phosphate" FT /ligand_id="ChEBI:CHEBI:61548" FT /evidence="ECO:0000269|PubMed:12498887, FT ECO:0000269|PubMed:20077550, ECO:0007744|PDB:1GZ5, FT ECO:0007744|PDB:2WTX" FT BINDING 131 FT /ligand="D-glucose 6-phosphate" FT /ligand_id="ChEBI:CHEBI:61548" FT /evidence="ECO:0000269|PubMed:12498887, FT ECO:0000269|PubMed:20077550, ECO:0007744|PDB:1GZ5, FT ECO:0007744|PDB:2WTX" FT BINDING 263 FT /ligand="UDP-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:58885" FT /evidence="ECO:0000305|PubMed:12498887, FT ECO:0000305|PubMed:14570926, ECO:0000305|PubMed:20077550, FT ECO:0007744|PDB:1GZ5, ECO:0007744|PDB:1UQT, FT ECO:0007744|PDB:1UQU, ECO:0007744|PDB:2WTX" FT BINDING 268 FT /ligand="UDP-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:58885" FT /evidence="ECO:0000305|PubMed:12498887, FT ECO:0000305|PubMed:14570926, ECO:0000305|PubMed:20077550, FT ECO:0007744|PDB:1GZ5, ECO:0007744|PDB:1UQT, FT ECO:0007744|PDB:1UQU, ECO:0007744|PDB:2WTX" FT BINDING 301 FT /ligand="D-glucose 6-phosphate" FT /ligand_id="ChEBI:CHEBI:61548" FT /evidence="ECO:0000269|PubMed:12498887, FT ECO:0000269|PubMed:20077550, ECO:0007744|PDB:1GZ5, FT ECO:0007744|PDB:2WTX" FT BINDING 340 FT /ligand="UDP-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:58885" FT /evidence="ECO:0000305|PubMed:12498887, FT ECO:0000305|PubMed:14570926, ECO:0000305|PubMed:20077550, FT ECO:0007744|PDB:1GZ5, ECO:0007744|PDB:1UQT, FT ECO:0007744|PDB:1UQU, ECO:0007744|PDB:2WTX" FT BINDING 366..370 FT /ligand="UDP-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:58885" FT /evidence="ECO:0000305|PubMed:12498887, FT ECO:0000305|PubMed:14570926, ECO:0000305|PubMed:20077550, FT ECO:0007744|PDB:1GZ5, ECO:0007744|PDB:1UQT, FT ECO:0007744|PDB:1UQU, ECO:0007744|PDB:2WTX" FT SITE 86 FT /note="Involved in alpha anomer selectivity" FT /evidence="ECO:0000305|PubMed:12498887" FT SITE 156 FT /note="Involved in alpha anomer selectivity" FT /evidence="ECO:0000305|PubMed:12498887" FT STRAND 4..10 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 16..18 FT /evidence="ECO:0007829|PDB:1GZ5" FT HELIX 23..35 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 37..47 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 54..58 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 61..68 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 70..76 FT /evidence="ECO:0007829|PDB:1UQT" FT TURN 77..79 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 80..83 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 85..89 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 93..95 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 100..117 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 118..120 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 126..131 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 132..134 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 137..143 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 150..153 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 161..164 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 170..177 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 180..187 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 188..201 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 204..207 FT /evidence="ECO:0007829|PDB:1UQT" FT TURN 208..210 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 211..214 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 217..223 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 230..238 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 243..251 FT /evidence="ECO:0007829|PDB:1UQT" FT TURN 252..254 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 256..261 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 266..268 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 270..283 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 285..287 FT /evidence="ECO:0007829|PDB:1UQT" FT TURN 288..290 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 291..296 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 305..325 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 332..336 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 342..351 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 353..357 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 360..363 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 366..374 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 382..386 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 390..393 FT /evidence="ECO:0007829|PDB:1UQT" FT STRAND 398..401 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 406..417 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 421..437 FT /evidence="ECO:0007829|PDB:1UQT" FT HELIX 440..453 FT /evidence="ECO:0007829|PDB:1UQT" SQ SEQUENCE 474 AA; 53611 MW; 63FFB6F938056F95 CRC64; MSRLVVVSNR IAPPDEHAAS AGGLAVGILG ALKAAGGLWF GWSGETGNED QPLKKVKKGN ITWASFNLSE QDLDEYYNQF SNAVLWPAFH YRLDLVQFQR PAWDGYLRVN ALLADKLLPL LQDDDIIWIH DYHLLPFAHE LRKRGVNNRI GFFLHIPFPT PEIFNALPTY DTLLEQLCDY DLLGFQTEND RLAFLDCLSN LTRVTTRSAK SHTAWGKAFR TEVYPIGIEP KEIAKQAAGP LPPKLAQLKA ELKNVQNIFS VERLDYSKGL PERFLAYEAL LEKYPQHHGK IRYTQIAPTS RGDVQAYQDI RHQLENEAGR INGKYGQLGW TPLYYLNQHF DRKLLMKIFR YSDVGLVTPL RDGMNLVAKE YVAAQDPANP GVLVLSQFAG AANELTSALI VNPYDRDEVA AALDRALTMS LAERISRHAE MLDVIVKNDI NHWQECFISD LKQIVPRSAE SQQRDKVATF PKLA //