SYT_ECOLI
ID SYT_ECOLI Reviewed; 642 AA.
AC P0A8M3; P00955; P78166; P78241;
DT 21-JUL-1986, integrated into UniProtKB/Swiss-Prot.
DT 07-JUN-2005, sequence version 1.
DT 03-AUG-2022, entry version 150.
DE RecName: Full=Threonine--tRNA ligase {ECO:0000255|HAMAP-Rule:MF_00184};
DE EC=6.1.1.3 {ECO:0000255|HAMAP-Rule:MF_00184, ECO:0000269|PubMed:10319817};
DE AltName: Full=Threonyl-tRNA synthetase {ECO:0000255|HAMAP-Rule:MF_00184};
DE Short=ThrRS {ECO:0000255|HAMAP-Rule:MF_00184};
GN Name=thrS {ECO:0000255|HAMAP-Rule:MF_00184};
GN OrderedLocusNames=b1719, JW1709;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=6353409; DOI=10.1073/pnas.80.20.6152;
RA Mayaux J.-F., Fayat G., Fromant M., Springer M., Grunberg-Manago M.,
RA Blanquet S.;
RT "Structural and transcriptional evidence for related thrS and infC
RT expression.";
RL Proc. Natl. Acad. Sci. U.S.A. 80:6152-6156(1983).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=9097039; DOI=10.1093/dnares/3.6.363;
RA Aiba H., Baba T., Fujita K., Hayashi K., Inada T., Isono K., Itoh T.,
RA Kasai H., Kashimoto K., Kimura S., Kitakawa M., Kitagawa M., Makino K.,
RA Miki T., Mizobuchi K., Mori H., Mori T., Motomura K., Nakade S.,
RA Nakamura Y., Nashimoto H., Nishio Y., Oshima T., Saito N., Sampei G.,
RA Seki Y., Sivasundaram S., Tagami H., Takeda J., Takemoto K., Takeuchi Y.,
RA Wada C., Yamamoto Y., Horiuchi T.;
RT "A 570-kb DNA sequence of the Escherichia coli K-12 genome corresponding to
RT the 28.0-40.1 min region on the linkage map.";
RL DNA Res. 3:363-377(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 1-47, AND MECHANISM OF TRANSLATION
RP REGULATION.
RX PubMed=3086882; DOI=10.1073/pnas.83.12.4384;
RA Springer M., Graffe M., Butler J.S., Grunberg-Manago M.;
RT "Genetic definition of the translational operator of the threonine-tRNA
RT ligase gene in Escherichia coli.";
RL Proc. Natl. Acad. Sci. U.S.A. 83:4384-4388(1986).
RN [6]
RP MECHANISM OF TRANSLATION REGULATION, AND RNA-BINDING.
RX PubMed=2254931; DOI=10.1016/s0022-2836(05)80321-3;
RA Moine H., Romby P., Springer M., Grunberg-Manago M., Ebel J.P.,
RA Ehresmann B., Ehresmann C.;
RT "Escherichia coli threonyl-tRNA synthetase and tRNA(Thr) modulate the
RT binding of the ribosome to the translational initiation site of the thrS
RT mRNA.";
RL J. Mol. Biol. 216:299-310(1990).
RN [7]
RP IDENTIFICATION BY 2D-GEL.
RX PubMed=9298644; DOI=10.1002/elps.1150180805;
RA VanBogelen R.A., Abshire K.Z., Moldover B., Olson E.R., Neidhardt F.C.;
RT "Escherichia coli proteome analysis using the gene-protein database.";
RL Electrophoresis 18:1243-1251(1997).
RN [8]
RP FUNCTION IN AMINOACYLATION AND EDITING, AND DOMAIN.
RX PubMed=15079065; DOI=10.1073/pnas.0401530101;
RA Beebe K., Merriman E., Ribas De Pouplana L., Schimmel P.;
RT "A domain for editing by an archaebacterial tRNA synthetase.";
RL Proc. Natl. Acad. Sci. U.S.A. 101:5958-5963(2004).
RN [9]
RP FUNCTION, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, DISRUPTION
RP PHENOTYPE, ANTIBIOTIC RESISTANCE, AND MUTAGENESIS OF PRO-296; THR-307;
RP HIS-309; HIS-337 AND LEU-489.
RX PubMed=15507440; DOI=10.1074/jbc.m411039200;
RA Ruan B., Bovee M.L., Sacher M., Stathopoulos C., Poralla K.,
RA Francklyn C.S., Soell D.;
RT "A unique hydrophobic cluster near the active site contributes to
RT differences in borrelidin inhibition among threonyl-tRNA synthetases.";
RL J. Biol. Chem. 280:571-577(2005).
RN [10]
RP FUNCTION, REACTION MECHANISM, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP MUTAGENESIS OF HIS-309; ARG-363; GLN-381; LYS-465 AND GLN-479.
RX PubMed=18997014; DOI=10.1073/pnas.0804247105;
RA Minajigi A., Francklyn C.S.;
RT "RNA-assisted catalysis in a protein enzyme: The 2'-hydroxyl of tRNA(Thr)
RT A76 promotes aminoacylation by threonyl-tRNA synthetase.";
RL Proc. Natl. Acad. Sci. U.S.A. 105:17748-17753(2008).
RN [11]
RP ACETYLATION [LARGE SCALE ANALYSIS] AT LYS-286, AND IDENTIFICATION BY MASS
RP SPECTROMETRY.
RC STRAIN=K12 / JW1106, and K12 / MG1655 / ATCC 47076;
RX PubMed=18723842; DOI=10.1074/mcp.m800187-mcp200;
RA Zhang J., Sprung R., Pei J., Tan X., Kim S., Zhu H., Liu C.F.,
RA Grishin N.V., Zhao Y.;
RT "Lysine acetylation is a highly abundant and evolutionarily conserved
RT modification in Escherichia coli.";
RL Mol. Cell. Proteomics 8:215-225(2009).
RN [12] {ECO:0007744|PDB:1QF6}
RP X-RAY CRYSTALLOGRAPHY (2.9 ANGSTROMS) IN COMPLEX WITH COGNATE TRNA; AMP AND
RP ZINC, FUNCTION, SUBUNIT, DOMAIN, AND MUTAGENESIS OF CYS-334; HIS-385 AND
RP HIS-511.
RX PubMed=10319817; DOI=10.1016/s0092-8674(00)80746-1;
RA Sankaranarayanan R., Dock-Bregeon A.-C., Romby P., Caillet J., Springer M.,
RA Rees B., Ehresmann C., Ehresmann B., Moras D.;
RT "The structure of threonyl-tRNA synthetase-tRNA(Thr) complex enlightens its
RT repressor activity and reveals an essential zinc ion in the active site.";
RL Cell 97:371-381(1999).
RN [13] {ECO:0007744|PDB:1FYF}
RP X-RAY CRYSTALLOGRAPHY (1.65 ANGSTROMS) OF 242-642 IN COMPLEX WITH EDITING
RP SUBSTRATE ANALOG AND ZINC, FUNCTION, SUBUNIT, AND MUTAGENESIS OF
RP 73-HIS--HIS-77 AND ASP-180.
RX PubMed=11136973; DOI=10.1016/s0092-8674(00)00191-4;
RA Dock-Bregeon A., Sankaranarayanan R., Romby P., Caillet J., Springer M.,
RA Rees B., Francklyn C.S., Ehresmann C., Moras D.;
RT "Transfer RNA-mediated editing in threonyl-tRNA synthetase. The class II
RT solution to the double discrimination problem.";
RL Cell 103:877-884(2000).
RN [14] {ECO:0007744|PDB:1EVK, ECO:0007744|PDB:1EVL}
RP X-RAY CRYSTALLOGRAPHY (1.55 ANGSTROMS) OF 242-642 IN COMPLEX WITH THREONINE
RP AND ITS ANALOG AND ZINC, FUNCTION, CATALYTIC ACTIVITY, AND SUBUNIT.
RX PubMed=10881191; DOI=10.1038/75856;
RA Sankaranarayanan R., Dock-Bregeon A.-C., Rees B., Bovee M., Caillet J.,
RA Romby P., Francklyn C.S., Moras D.;
RT "Zinc ion mediated amino acid discrimination by threonyl-tRNA synthetase.";
RL Nat. Struct. Biol. 7:461-465(2000).
RN [15] {ECO:0007744|PDB:1KOG}
RP X-RAY CRYSTALLOGRAPHY (3.50 ANGSTROMS) OF 242-642 IN COMPLEX WITH OPERATOR
RP FRAGMENT OF MRNA AND ZINC, FUNCTION, AND SUBUNIT.
RX PubMed=11953757; DOI=10.1038/nsb789;
RA Torres-Larios A., Dock-Bregeon A.C., Romby P., Rees B.,
RA Sankaranarayanan R., Caillet J., Springer M., Ehresmann C., Ehresmann B.,
RA Moras D.;
RT "Structural basis of translational control by Escherichia coli threonyl
RT tRNA synthetase.";
RL Nat. Struct. Biol. 9:343-347(2002).
RN [16] {ECO:0007744|PDB:1TJE, ECO:0007744|PDB:1TKE, ECO:0007744|PDB:1TKG, ECO:0007744|PDB:1TKY}
RP X-RAY CRYSTALLOGRAPHY (1.46 ANGSTROMS) OF 1-224 (EDITING DOMAIN) IN COMPLEX
RP WITH EDITING ANALOGS, FUNCTION, POSSIBLE EDITING REACTION MECHANISM,
RP DOMAIN, AND MUTAGENESIS OF LYS-156; CYS-182 AND HIS-186.
RX PubMed=15525511; DOI=10.1016/j.molcel.2004.10.002;
RA Dock-Bregeon A.C., Rees B., Torres-Larios A., Bey G., Caillet J., Moras D.;
RT "Achieving error-free translation; the mechanism of proofreading of
RT threonyl-tRNA synthetase at atomic resolution.";
RL Mol. Cell 16:375-386(2004).
RN [17] {ECO:0007744|PDB:4HWO, ECO:0007744|PDB:4HWP, ECO:0007744|PDB:4HWR, ECO:0007744|PDB:4HWS}
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) OF 242-642 IN COMPLEX WITH
RP INHIBITORS AND ZINC, SUBUNIT, AND BIOTECHNOLOGY.
RX PubMed=23362938; DOI=10.1021/jm301756m;
RA Teng M., Hilgers M.T., Cunningham M.L., Borchardt A., Locke J.B.,
RA Abraham S., Haley G., Kwan B.P., Hall C., Hough G.W., Shaw K.J., Finn J.;
RT "Identification of bacteria-selective threonyl-tRNA synthetase substrate
RT inhibitors by structure-based design.";
RL J. Med. Chem. 56:1748-1760(2013).
RN [18] {ECO:0007744|PDB:4P3O, ECO:0007744|PDB:4P3P}
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 242-642 IN COMPLEX WITH
RP BORRELIDIN AND ZINC, ACTIVITY REGULATION, AND SUBUNIT.
RX PubMed=25824639; DOI=10.1038/ncomms7402;
RA Fang P., Yu X., Jeong S.J., Mirando A., Chen K., Chen X., Kim S.,
RA Francklyn C.S., Guo M.;
RT "Structural basis for full-spectrum inhibition of translational functions
RT on a tRNA synthetase.";
RL Nat. Commun. 6:6402-6402(2015).
CC -!- FUNCTION: Catalyzes the attachment of threonine to tRNA(Thr) in a two-
CC step reaction: L-threonine is first activated by ATP to form Thr-AMP
CC and then transferred to the acceptor end of tRNA(Thr) (PubMed:15079065,
CC PubMed:10881191, PubMed:18997014). The rate-limiting step is amino acid
CC activation in the presence of tRNA (PubMed:18997014). The 2'-OH of the
CC acceptor base (adenine 76, A76) of tRNA(Thr) and His-309 collaborate to
CC transfer L-Thr to the tRNA; substitution of 2'-OH of A76 with hydrogen
CC or fluorine decreases transfer efficiency 760 and 100-fold respectively
CC (PubMed:18997014). The zinc ion in the active site discriminates
CC against charging of the isosteric amino acid valine (PubMed:10881191).
CC Also activates L-serine, but does not detectably transfer it to
CC tRNA(Thr) (PubMed:15079065). Edits incorrectly charged L-seryl-
CC tRNA(Thr) via its editing domain (PubMed:15079065, PubMed:11136973,
CC PubMed:15525511), in a post-transfer reaction probably via water-
CC mediated hydrolysis (PubMed:15525511). {ECO:0000269|PubMed:10319817,
CC ECO:0000269|PubMed:10881191, ECO:0000269|PubMed:11136973,
CC ECO:0000269|PubMed:15079065, ECO:0000269|PubMed:15525511,
CC ECO:0000269|PubMed:18997014}.
CC -!- FUNCTION: ThrS is also a translational repressor protein, it controls
CC binds its own mRNA in the operator region upstream of the start codon
CC (PubMed:3086882). The mRNA region upstream of the start codon has a
CC tRNA-like secondary structure; mRNA and tRNA compete for binding to
CC ThrRS (PubMed:2254931). ThrRS represses translation by preventing the
CC ribosome from to mRNA, and tRNA(Thr) acts as an antirepressor allowing
CC fine level control of enzyme synthesis (PubMed:2254931). X-ray
CC structures prove that operator mRNA and tRNA bind to overlapping sites
CC in the protein (PubMed:10319817, PubMed:11953757).
CC {ECO:0000269|PubMed:10319817, ECO:0000269|PubMed:11953757,
CC ECO:0000269|PubMed:2254931, ECO:0000269|PubMed:3086882}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + L-threonine + tRNA(Thr) = AMP + diphosphate + H(+) + L-
CC threonyl-tRNA(Thr); Xref=Rhea:RHEA:24624, Rhea:RHEA-COMP:9670,
CC Rhea:RHEA-COMP:9704, ChEBI:CHEBI:15378, ChEBI:CHEBI:30616,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:57926, ChEBI:CHEBI:78442,
CC ChEBI:CHEBI:78534, ChEBI:CHEBI:456215; EC=6.1.1.3;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00184,
CC ECO:0000269|PubMed:10319817, ECO:0000269|PubMed:10881191,
CC ECO:0000269|PubMed:18997014};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00184};
CC Note=Binds 1 zinc ion per subunit. It helps recognize and select the
CC amino acid substrate, and thus has neither a purely catalytic or
CC structural role (PubMed:10881191). {ECO:0000269|PubMed:10319817,
CC ECO:0000269|PubMed:10881191, ECO:0000269|PubMed:11136973,
CC ECO:0000269|PubMed:11953757, ECO:0000269|PubMed:23362938,
CC ECO:0000269|PubMed:25824639};
CC -!- ACTIVITY REGULATION: Inhibited non-competitively by borrelidin (BN, KI
CC is 3.7 nM) which binds in a 1:1 stoichiometry, inhibiting L-thr
CC activation (PubMed:15507440). BN binds to 4 distinct subsites in the
CC protein, preventing binding of all 3 substrates; BN also inhibits human
CC ThrRS, and thus it is not useful as an antibiotic (PubMed:25824639).
CC {ECO:0000269|PubMed:15507440, ECO:0000269|PubMed:25824639}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=94 uM for ATP {ECO:0000269|PubMed:15507440};
CC KM=110 uM for L-threonine activation {ECO:0000269|PubMed:10881191,
CC ECO:0000269|PubMed:15507440};
CC KM=120 uM for L-threonine activation {ECO:0000269|PubMed:18997014};
CC KM=0.86 uM for L-threonine aminoacylation
CC {ECO:0000269|PubMed:18997014};
CC KM=1.95 mM for beta-hydroxynorvaline activation
CC {ECO:0000269|PubMed:10881191};
CC KM=81.5 mM for L-serine activation {ECO:0000269|PubMed:10881191};
CC Note=kcat is 36, 22 and 26 sec(-1) for L-threonine, beta-
CC hydroxynorvaline and L-serine respectively.
CC {ECO:0000269|PubMed:10881191};
CC -!- SUBUNIT: Homodimer (PubMed:10319817, PubMed:11136973, PubMed:10881191,
CC PubMed:11953757, PubMed:23362938, PubMed:25824639); binds 2 tRNA(Thr)
CC per homodimer, each tRNA contacts both monomers and makes specific
CC contacts with the anticodon and acceptor stems (PubMed:10319817).
CC {ECO:0000269|PubMed:10319817, ECO:0000269|PubMed:10881191,
CC ECO:0000269|PubMed:11136973, ECO:0000269|PubMed:11953757,
CC ECO:0000269|PubMed:23362938, ECO:0000269|PubMed:25824639}.
CC -!- INTERACTION:
CC P0A8M3; P0A7L0: rplA; NbExp=3; IntAct=EBI-551254, EBI-543771;
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_00184}.
CC -!- DOMAIN: The protein structure shows 2 N-terminal domains, the central
CC catalytic and C-terminal domain (PubMed:10319817). The C-terminal
CC domain recognizes the anticodon region of the tRNA while the acceptor
CC arm is sandwiched between the N-terminal domains and the catalytic
CC domain (PubMed:10319817). The N-terminal also contributes to the
CC precise recognition of tRNA(Thr) (PubMed:10319817). The editing domain
CC encompasses approximately residues 62-224; when it is removed the
CC protein mischarges tRNA(Thr) with L-serine (PubMed:15079065,
CC PubMed:11136973). {ECO:0000269|PubMed:11136973,
CC ECO:0000269|PubMed:15525511, ECO:0000305|PubMed:10319817,
CC ECO:0000305|PubMed:15079065}.
CC -!- DISRUPTION PHENOTYPE: Essential, it cannot be deleted.
CC {ECO:0000269|PubMed:15507440}.
CC -!- BIOTECHNOLOGY: A number of inhibitors with high affinity for bacterial
CC ThrRS and less affinity for human ThrRS have been identified that might
CC make good antibiotics. {ECO:0000269|PubMed:23362938}.
CC -!- SIMILARITY: Belongs to the class-II aminoacyl-tRNA synthetase family.
CC {ECO:0000255|HAMAP-Rule:MF_00184}.
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DR EMBL; V00291; CAA23560.1; -; Genomic_DNA.
DR EMBL; U00096; AAC74789.1; -; Genomic_DNA.
DR EMBL; AP009048; BAA15498.1; -; Genomic_DNA.
DR EMBL; M13549; AAA24674.1; -; Genomic_DNA.
DR PIR; G64930; SYECTT.
DR RefSeq; NP_416234.1; NC_000913.3.
DR RefSeq; WP_001144202.1; NZ_SSZK01000001.1.
DR PDB; 1EVK; X-ray; 2.00 A; A/B=242-642.
DR PDB; 1EVL; X-ray; 1.55 A; A/B/C/D=242-642.
DR PDB; 1FYF; X-ray; 1.65 A; A/B=242-642.
DR PDB; 1KOG; X-ray; 3.50 A; A/B/C/D/E/F/G/H=242-642.
DR PDB; 1QF6; X-ray; 2.90 A; A=1-642.
DR PDB; 1TJE; X-ray; 1.50 A; A=1-224.
DR PDB; 1TKE; X-ray; 1.46 A; A=1-224.
DR PDB; 1TKG; X-ray; 1.50 A; A=1-224.
DR PDB; 1TKY; X-ray; 1.48 A; A=1-224.
DR PDB; 4HWO; X-ray; 1.91 A; A/B=242-642.
DR PDB; 4HWP; X-ray; 1.81 A; A/B=242-642.
DR PDB; 4HWR; X-ray; 1.90 A; A/B=242-642.
DR PDB; 4HWS; X-ray; 1.70 A; A/B=242-642.
DR PDB; 4P3O; X-ray; 2.50 A; A/B=242-642.
DR PDB; 4P3P; X-ray; 2.10 A; A/B=242-642.
DR PDBsum; 1EVK; -.
DR PDBsum; 1EVL; -.
DR PDBsum; 1FYF; -.
DR PDBsum; 1KOG; -.
DR PDBsum; 1QF6; -.
DR PDBsum; 1TJE; -.
DR PDBsum; 1TKE; -.
DR PDBsum; 1TKG; -.
DR PDBsum; 1TKY; -.
DR PDBsum; 4HWO; -.
DR PDBsum; 4HWP; -.
DR PDBsum; 4HWR; -.
DR PDBsum; 4HWS; -.
DR PDBsum; 4P3O; -.
DR PDBsum; 4P3P; -.
DR AlphaFoldDB; P0A8M3; -.
DR SMR; P0A8M3; -.
DR BioGRID; 4262193; 27.
DR BioGRID; 850582; 2.
DR DIP; DIP-35823N; -.
DR IntAct; P0A8M3; 50.
DR MINT; P0A8M3; -.
DR STRING; 511145.b1719; -.
DR MoonProt; P0A8M3; -.
DR iPTMnet; P0A8M3; -.
DR jPOST; P0A8M3; -.
DR PaxDb; P0A8M3; -.
DR PRIDE; P0A8M3; -.
DR EnsemblBacteria; AAC74789; AAC74789; b1719.
DR EnsemblBacteria; BAA15498; BAA15498; BAA15498.
DR GeneID; 58460397; -.
DR GeneID; 946222; -.
DR KEGG; ecj:JW1709; -.
DR KEGG; eco:b1719; -.
DR PATRIC; fig|1411691.4.peg.538; -.
DR EchoBASE; EB0994; -.
DR eggNOG; COG0441; Bacteria.
DR HOGENOM; CLU_008554_0_1_6; -.
DR InParanoid; P0A8M3; -.
DR OMA; FYYDFAY; -.
DR PhylomeDB; P0A8M3; -.
DR BioCyc; EcoCyc:THRS-MON; -.
DR BioCyc; MetaCyc:THRS-MON; -.
DR BRENDA; 6.1.1.3; 2026.
DR SABIO-RK; P0A8M3; -.
DR EvolutionaryTrace; P0A8M3; -.
DR PRO; PR:P0A8M3; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IDA:EcoliWiki.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0002161; F:aminoacyl-tRNA editing activity; IDA:EcoCyc.
DR GO; GO:0004812; F:aminoacyl-tRNA ligase activity; IDA:EcoliWiki.
DR GO; GO:0005524; F:ATP binding; IDA:EcoliWiki.
DR GO; GO:0048027; F:mRNA 5'-UTR binding; IDA:EcoCyc.
DR GO; GO:0000900; F:mRNA regulatory element binding translation repressor activity; IDA:EcoCyc.
DR GO; GO:0003723; F:RNA binding; IDA:EcoliWiki.
DR GO; GO:0004829; F:threonine-tRNA ligase activity; IDA:EcoCyc.
DR GO; GO:0000049; F:tRNA binding; IEA:UniProtKB-KW.
DR GO; GO:0008270; F:zinc ion binding; IDA:EcoCyc.
DR GO; GO:0045947; P:negative regulation of translational initiation; IMP:EcoCyc.
DR GO; GO:0006417; P:regulation of translation; IDA:EcoCyc.
DR GO; GO:0046677; P:response to antibiotic; IEA:UniProtKB-KW.
DR GO; GO:0006435; P:threonyl-tRNA aminoacylation; IDA:EcoCyc.
DR GO; GO:0043039; P:tRNA aminoacylation; IDA:EcoliWiki.
DR GO; GO:0006418; P:tRNA aminoacylation for protein translation; IDA:EcoliWiki.
DR CDD; cd00771; ThrRS_core; 1.
DR Gene3D; 3.10.20.30; -; 1.
DR Gene3D; 3.30.930.10; -; 1.
DR Gene3D; 3.40.50.800; -; 1.
DR HAMAP; MF_00184; Thr_tRNA_synth; 1.
DR InterPro; IPR002314; aa-tRNA-synt_IIb.
DR InterPro; IPR006195; aa-tRNA-synth_II.
DR InterPro; IPR045864; aa-tRNA-synth_II/BPL/LPL.
DR InterPro; IPR004154; Anticodon-bd.
DR InterPro; IPR036621; Anticodon-bd_dom_sf.
DR InterPro; IPR012675; Beta-grasp_dom_sf.
DR InterPro; IPR004095; TGS.
DR InterPro; IPR012676; TGS-like.
DR InterPro; IPR002320; Thr-tRNA-ligase_IIa.
DR InterPro; IPR018163; Thr/Ala-tRNA-synth_IIc_edit.
DR InterPro; IPR033728; ThrRS_core.
DR InterPro; IPR012947; tRNA_SAD.
DR PANTHER; PTHR11451; PTHR11451; 1.
DR Pfam; PF03129; HGTP_anticodon; 1.
DR Pfam; PF02824; TGS; 1.
DR Pfam; PF00587; tRNA-synt_2b; 1.
DR Pfam; PF07973; tRNA_SAD; 1.
DR PRINTS; PR01047; TRNASYNTHTHR.
DR SMART; SM00863; tRNA_SAD; 1.
DR SUPFAM; SSF55186; SSF55186; 1.
DR SUPFAM; SSF55681; SSF55681; 1.
DR SUPFAM; SSF81271; SSF81271; 1.
DR TIGRFAMs; TIGR00418; thrS; 1.
DR PROSITE; PS50862; AA_TRNA_LIGASE_II; 1.
DR PROSITE; PS51880; TGS; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acetylation; Aminoacyl-tRNA synthetase;
KW Antibiotic resistance; ATP-binding; Cytoplasm; Ligase; Metal-binding;
KW Nucleotide-binding; Protein biosynthesis; Reference proteome; Repressor;
KW RNA-binding; Translation regulation; tRNA-binding; Zinc.
FT CHAIN 1..642
FT /note="Threonine--tRNA ligase"
FT /id="PRO_0000100973"
FT DOMAIN 1..61
FT /note="TGS"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01228"
FT REGION 1..224
FT /note="Correctly edits mischarged seryl-tRNA(Thr)"
FT /evidence="ECO:0000269|PubMed:15525511"
FT REGION 1..62
FT /note="N1 domain"
FT /evidence="ECO:0000305|PubMed:10319817"
FT REGION 2..241
FT /note="N-terminal region which includes the editing domain,
FT important for catalytic efficiency, its loss increases
FT mischarging with L-serine, deacylation of incorrectly
FT charged tRNA no longer occurs, partially complements a
FT deletion strain"
FT /evidence="ECO:0000269|PubMed:10319817"
FT REGION 63..224
FT /note="N2 domain, the editing domain"
FT /evidence="ECO:0000305|PubMed:10319817"
FT REGION 200..219
FT /note="tRNA acceptor stem binding"
FT /evidence="ECO:0000269|PubMed:10319817"
FT REGION 243..534
FT /note="Catalytic"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00184,
FT ECO:0000305|PubMed:10319817"
FT REGION 535..642
FT /note="Anticodon recognition"
FT /evidence="ECO:0000305|PubMed:10319817"
FT BINDING 246..249
FT /ligand="mRNA"
FT /ligand_id="ChEBI:CHEBI:33699"
FT /ligand_part="mRNA operator"
FT /evidence="ECO:0000269|PubMed:11953757"
FT BINDING 309
FT /ligand="tRNA(Thr)"
FT /ligand_id="ChEBI:CHEBI:29180"
FT /ligand_part="AMP 3'-end residue"
FT /ligand_part_id="ChEBI:CHEBI:78442"
FT /evidence="ECO:0000269|PubMed:10319817,
FT ECO:0007744|PDB:1QF6"
FT BINDING 313..317
FT /ligand="tRNA"
FT /ligand_id="ChEBI:CHEBI:17843"
FT /ligand_part="tRNA 3'-terminal nucleotidyl-cytidyl-cytidyl-
FT adenosine residue"
FT /ligand_part_id="ChEBI:CHEBI:83071"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 325
FT /ligand="tRNA(Thr)"
FT /ligand_id="ChEBI:CHEBI:29180"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 334
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00184,
FT ECO:0000269|PubMed:10319817, ECO:0000269|PubMed:10881191,
FT ECO:0000269|PubMed:11136973, ECO:0000269|PubMed:11953757"
FT BINDING 342..349
FT /ligand="mRNA"
FT /ligand_id="ChEBI:CHEBI:33699"
FT /ligand_part="mRNA operator"
FT /evidence="ECO:0000269|PubMed:11953757"
FT BINDING 348..349
FT /ligand="tRNA(Thr)"
FT /ligand_id="ChEBI:CHEBI:29180"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 363..365
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000269|PubMed:10319817,
FT ECO:0007744|PDB:1QF6"
FT BINDING 363
FT /ligand="tRNA"
FT /ligand_id="ChEBI:CHEBI:17843"
FT /ligand_part="tRNA 3'-terminal nucleotidyl-cytidyl-cytidyl-
FT adenosine residue"
FT /ligand_part_id="ChEBI:CHEBI:83071"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 375
FT /ligand="tRNA"
FT /ligand_id="ChEBI:CHEBI:17843"
FT /ligand_part="tRNA 3'-terminal nucleotidyl-cytidyl-cytidyl-
FT adenosine residue"
FT /ligand_part_id="ChEBI:CHEBI:83071"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 376
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000269|PubMed:10319817,
FT ECO:0007744|PDB:1QF6"
FT BINDING 379
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 381
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000269|PubMed:10319817,
FT ECO:0007744|PDB:1QF6"
FT BINDING 385
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00184,
FT ECO:0000269|PubMed:10319817, ECO:0000269|PubMed:10881191,
FT ECO:0000269|PubMed:11136973, ECO:0000269|PubMed:11953757"
FT BINDING 462
FT /ligand="tRNA"
FT /ligand_id="ChEBI:CHEBI:17843"
FT /ligand_part="tRNA 3'-terminal nucleotidyl-cytidyl-cytidyl-
FT adenosine residue"
FT /ligand_part_id="ChEBI:CHEBI:83071"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 479..480
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000269|PubMed:10319817,
FT ECO:0007744|PDB:1QF6"
FT BINDING 484
FT /ligand="tRNA"
FT /ligand_id="ChEBI:CHEBI:17843"
FT /ligand_part="tRNA 3'-terminal nucleotidyl-cytidyl-cytidyl-
FT adenosine residue"
FT /ligand_part_id="ChEBI:CHEBI:83071"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 489..503
FT /ligand="mRNA"
FT /ligand_id="ChEBI:CHEBI:33699"
FT /ligand_part="mRNA operator"
FT /evidence="ECO:0000269|PubMed:11953757"
FT BINDING 511
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00184,
FT ECO:0000269|PubMed:10319817, ECO:0000269|PubMed:10881191,
FT ECO:0000269|PubMed:11136973"
FT BINDING 517
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000269|PubMed:10319817,
FT ECO:0007744|PDB:1QF6"
FT BINDING 520
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 547..549
FT /ligand="mRNA"
FT /ligand_id="ChEBI:CHEBI:33699"
FT /ligand_part="mRNA operator"
FT /evidence="ECO:0000269|PubMed:11953757"
FT BINDING 547..549
FT /ligand="tRNA"
FT /ligand_id="ChEBI:CHEBI:17843"
FT /ligand_part="anticodon region of tRNA"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 575..586
FT /ligand="mRNA"
FT /ligand_id="ChEBI:CHEBI:33699"
FT /ligand_part="mRNA operator"
FT /evidence="ECO:0000269|PubMed:11953757"
FT BINDING 575..583
FT /ligand="tRNA"
FT /ligand_id="ChEBI:CHEBI:17843"
FT /ligand_part="anticodon region of tRNA"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 589
FT /ligand="tRNA"
FT /ligand_id="ChEBI:CHEBI:17843"
FT /ligand_part="anticodon region of tRNA"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 595..600
FT /ligand="mRNA"
FT /ligand_id="ChEBI:CHEBI:33699"
FT /ligand_part="mRNA operator"
FT /evidence="ECO:0000269|PubMed:11953757"
FT BINDING 595..600
FT /ligand="tRNA"
FT /ligand_id="ChEBI:CHEBI:17843"
FT /ligand_part="anticodon region of tRNA"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 609
FT /ligand="mRNA"
FT /ligand_id="ChEBI:CHEBI:33699"
FT /ligand_part="mRNA operator"
FT /evidence="ECO:0000269|PubMed:11953757"
FT BINDING 609
FT /ligand="tRNA"
FT /ligand_id="ChEBI:CHEBI:17843"
FT /ligand_part="anticodon region of tRNA"
FT /evidence="ECO:0000269|PubMed:10319817"
FT BINDING 615
FT /ligand="mRNA"
FT /ligand_id="ChEBI:CHEBI:33699"
FT /ligand_part="mRNA operator"
FT /evidence="ECO:0000269|PubMed:11953757"
FT MOD_RES 286
FT /note="N6-acetyllysine"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00184,
FT ECO:0000269|PubMed:18723842"
FT MUTAGEN 73..77
FT /note="HSCAH->ASCAA: No longer edits mischarged L-seryl-
FT tRNA(Thr), mischarges tRNA(Thr) with L-serine, correct
FT acylation is unaffected."
FT /evidence="ECO:0000269|PubMed:11136973"
FT MUTAGEN 156
FT /note="K->A: Mischarges tRNA(Thr) with L-serine."
FT /evidence="ECO:0000269|PubMed:15525511"
FT MUTAGEN 180
FT /note="D->A: No longer edits mischarged L-seryl-tRNA(Thr),
FT mischarges tRNA(Thr) with L-serine, correct acylation is
FT unaffected."
FT /evidence="ECO:0000269|PubMed:11136973"
FT MUTAGEN 182
FT /note="C->A: Very high mischarging of tRNA(Thr) with L-
FT serine."
FT /evidence="ECO:0000269|PubMed:15525511"
FT MUTAGEN 186
FT /note="H->A: Mischarges tRNA(Thr) with L-serine."
FT /evidence="ECO:0000269|PubMed:15525511"
FT MUTAGEN 296
FT /note="P->S: Confers resistance to borrelidin (BN); KM for
FT L-Thr is unchanged, KM for ATP increases to 187 uM, KI for
FT BN increases to 4.5 nM."
FT /evidence="ECO:0000269|PubMed:15507440"
FT MUTAGEN 307
FT /note="T->A: KI for BN increases 10-fold, no change in
FT aminoacylation activity."
FT /evidence="ECO:0000269|PubMed:15507440"
FT MUTAGEN 309
FT /note="H->A: 10-fold increase in KM for Thr for activation,
FT 240-fold decrease in aminoacyl transfer. Cells have a long
FT lag phase and reach stationary phase at a lower cell
FT density. KI for BN increases 1000-fold, supports growth in
FT the presence of BN."
FT /evidence="ECO:0000269|PubMed:15507440,
FT ECO:0000269|PubMed:18997014"
FT MUTAGEN 334
FT /note="C->S: Does not complement a deletion strain."
FT /evidence="ECO:0000269|PubMed:10319817"
FT MUTAGEN 337
FT /note="H->A: KI for BN increases 12-fold, no change in
FT aminoacylation activity, supports growth in the presence of
FT BN."
FT /evidence="ECO:0000269|PubMed:15507440"
FT MUTAGEN 363
FT /note="R->A: 700-fold decrease in kcat for Thr activation,
FT 1000-fold decrease in kcat of aminoacylation, no change in
FT KM."
FT /evidence="ECO:0000269|PubMed:18997014"
FT MUTAGEN 381
FT /note="Q->A: 100-fold increase in KM for Thr for
FT activation."
FT /evidence="ECO:0000269|PubMed:18997014"
FT MUTAGEN 385
FT /note="H->A,N: Does not complement a deletion strain."
FT /evidence="ECO:0000269|PubMed:10319817"
FT MUTAGEN 465
FT /note="K->A: 35-fold decrease in kcat for Thr activation,
FT 570-fold decrease in kcat of aminoacylation, no change in
FT KM."
FT /evidence="ECO:0000269|PubMed:18997014"
FT MUTAGEN 479
FT /note="Q->A: Wild-type Thr activation and aminoacylation."
FT /evidence="ECO:0000269|PubMed:18997014"
FT MUTAGEN 489
FT /note="L->M: Confers resistance to borrelidin (BN); KM for
FT L-thr is unchanged, KM for ATP increases to 163 uM, KI for
FT BN increases to 7.8 nM, supports growth in the presence of
FT BN."
FT /evidence="ECO:0000269|PubMed:15507440"
FT MUTAGEN 489
FT /note="L->W: KI for BN increases 1500-fold, no change in
FT aminoacylation activity, supports growth in the presence of
FT BN."
FT /evidence="ECO:0000269|PubMed:15507440"
FT MUTAGEN 511
FT /note="H->A,N: Does not complement a deletion strain, has
FT dominant lethal effect in presence of wild-type gene,
FT probably due to repression of the wild-type gene."
FT /evidence="ECO:0000269|PubMed:10319817"
FT MUTAGEN 531
FT /note="G->GEGK: KI for BN increases 8-fold, decreases
FT aminoacylation activity, does not support growth in the
FT presence of BN."
FT /evidence="ECO:0000269|PubMed:15507440"
FT CONFLICT 195
FT /note="H -> R (in Ref. 1; CAA23560)"
FT /evidence="ECO:0000305"
FT STRAND 3..5
FT /evidence="ECO:0007829|PDB:1TKE"
FT STRAND 11..13
FT /evidence="ECO:0007829|PDB:1TKE"
FT HELIX 20..27
FT /evidence="ECO:0007829|PDB:1TKE"
FT HELIX 29..34
FT /evidence="ECO:0007829|PDB:1TKE"
FT STRAND 37..40
FT /evidence="ECO:0007829|PDB:1TKE"
FT STRAND 43..46
FT /evidence="ECO:0007829|PDB:1TKE"
FT STRAND 55..60
FT /evidence="ECO:0007829|PDB:1TKE"
FT HELIX 65..86
FT /evidence="ECO:0007829|PDB:1TKE"
FT STRAND 101..107
FT /evidence="ECO:0007829|PDB:1TKE"
FT HELIX 114..128
FT /evidence="ECO:0007829|PDB:1TKE"
FT STRAND 135..138
FT /evidence="ECO:0007829|PDB:1TKE"
FT HELIX 141..150
FT /evidence="ECO:0007829|PDB:1TKE"
FT HELIX 154..163
FT /evidence="ECO:0007829|PDB:1TKE"
FT STRAND 171..175
FT /evidence="ECO:0007829|PDB:1TKE"
FT STRAND 178..183
FT /evidence="ECO:0007829|PDB:1TKE"
FT HELIX 190..192
FT /evidence="ECO:0007829|PDB:1TKE"
FT STRAND 196..205
FT /evidence="ECO:0007829|PDB:1TKE"
FT HELIX 206..208
FT /evidence="ECO:0007829|PDB:1TKE"
FT STRAND 214..222
FT /evidence="ECO:0007829|PDB:1TKE"
FT HELIX 226..240
FT /evidence="ECO:0007829|PDB:1QF6"
FT HELIX 244..250
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 268..287
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 297..300
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 301..306
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 309..312
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 314..316
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 319..322
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 325..329
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 331..333
FT /evidence="ECO:0007829|PDB:1QF6"
FT HELIX 334..341
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 348..350
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 352..362
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 367..369
FT /evidence="ECO:0007829|PDB:1EVL"
FT TURN 372..374
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 377..388
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 390..392
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 393..409
FT /evidence="ECO:0007829|PDB:1EVL"
FT TURN 410..412
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 417..421
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 431..447
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 453..455
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 461..463
FT /evidence="ECO:0007829|PDB:4HWR"
FT STRAND 465..471
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 477..488
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 489..492
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 502..505
FT /evidence="ECO:0007829|PDB:1EVK"
FT STRAND 508..517
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 518..529
FT /evidence="ECO:0007829|PDB:1EVL"
FT TURN 535..537
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 542..548
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 549..551
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 552..564
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 569..572
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 574..576
FT /evidence="ECO:0007829|PDB:4P3O"
FT HELIX 578..587
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 591..596
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 598..603
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 605..610
FT /evidence="ECO:0007829|PDB:1EVL"
FT STRAND 615..620
FT /evidence="ECO:0007829|PDB:1EVL"
FT HELIX 621..633
FT /evidence="ECO:0007829|PDB:1EVL"
SQ SEQUENCE 642 AA; 74014 MW; 38D8913B19CDAB7B CRC64;
MPVITLPDGS QRHYDHAVSP MDVALDIGPG LAKACIAGRV NGELVDACDL IENDAQLSII
TAKDEEGLEI IRHSCAHLLG HAIKQLWPHT KMAIGPVIDN GFYYDVDLDR TLTQEDVEAL
EKRMHELAEK NYDVIKKKVS WHEARETFAN RGESYKVSIL DENIAHDDKP GLYFHEEYVD
MCRGPHVPNM RFCHHFKLMK TAGAYWRGDS NNKMLQRIYG TAWADKKALN AYLQRLEEAA
KRDHRKIGKQ LDLYHMQEEA PGMVFWHNDG WTIFRELEVF VRSKLKEYQY QEVKGPFMMD
RVLWEKTGHW DNYKDAMFTT SSENREYCIK PMNCPGHVQI FNQGLKSYRD LPLRMAEFGS
CHRNEPSGSL HGLMRVRGFT QDDAHIFCTE EQIRDEVNGC IRLVYDMYST FGFEKIVVKL
STRPEKRIGS DEMWDRAEAD LAVALEENNI PFEYQLGEGA FYGPKIEFTL YDCLDRAWQC
GTVQLDFSLP SRLSASYVGE DNERKVPVMI HRAILGSMER FIGILTEEFA GFFPTWLAPV
QVVIMNITDS QSEYVNELTQ KLSNAGIRVK ADLRNEKIGF KIREHTLRRV PYMLVCGDKE
VESGKVAVRT RRGKDLGSMD VNEVIEKLQQ EIRSRSLKQL EE
