TGT_ECOLI
ID TGT_ECOLI Reviewed; 375 AA.
AC P0A847; P19675; P19676; P78226; Q2MC20; Q47627;
DT 07-JUN-2005, integrated into UniProtKB/Swiss-Prot.
DT 07-JUN-2005, sequence version 1.
DT 03-AUG-2022, entry version 128.
DE RecName: Full=Queuine tRNA-ribosyltransferase {ECO:0000255|HAMAP-Rule:MF_00168};
DE EC=2.4.2.29 {ECO:0000255|HAMAP-Rule:MF_00168, ECO:0000269|PubMed:11714265, ECO:0000269|PubMed:12909636};
DE AltName: Full=Guanine insertion enzyme {ECO:0000255|HAMAP-Rule:MF_00168};
DE AltName: Full=tRNA-guanine transglycosylase {ECO:0000255|HAMAP-Rule:MF_00168};
GN Name=tgt {ECO:0000255|HAMAP-Rule:MF_00168};
GN OrderedLocusNames=b0406, JW0396;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PATHWAY.
RC STRAIN=K12;
RX PubMed=1706703; DOI=10.1128/jb.173.7.2256-2264.1991;
RA Reuter K., Slany R., Ullrich F., Kersten H.;
RT "Structure and organization of Escherichia coli genes involved in
RT biosynthesis of the deazaguanine derivative queuine, a nutrient factor for
RT eukaryotes.";
RL J. Bacteriol. 173:2256-2264(1991).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PROTEIN SEQUENCE OF 1-9.
RX PubMed=8323579; DOI=10.1006/jmbi.1993.1296;
RA Garcia G.A., Koch K.A., Chong S.;
RT "tRNA-guanine transglycosylase from Escherichia coli. Overexpression,
RT purification and quaternary structure.";
RL J. Mol. Biol. 231:489-497(1993).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RA Chung E., Allen E., Araujo R., Aparicio A.M., Davis K., Duncan M.,
RA Federspiel N., Hyman R., Kalman S., Komp C., Kurdi O., Lew H., Lin D.,
RA Namath A., Oefner P., Roberts D., Schramm S., Davis R.W.;
RT "Sequence of minutes 4-25 of Escherichia coli.";
RL Submitted (JAN-1997) to the EMBL/GenBank/DDBJ databases.
RN [4]
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 [5]
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 [6]
RP PRELIMINARY NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 141-375.
RC STRAIN=K12;
RX PubMed=2170107; DOI=10.1002/j.1460-2075.1990.tb07519.x;
RA Gardel C., Johnson K., Jacq A., Beckwith J.;
RT "The secD locus of E.coli codes for two membrane proteins required for
RT protein export.";
RL EMBO J. 9:3209-3216(1990).
RN [7]
RP ERRATUM OF PUBMED:2170107.
RX PubMed=2249673; DOI=10.1002/j.1460-2075.1990.tb07645.x;
RA Gardel C., Johnson K., Jacq A., Beckwith J.;
RL EMBO J. 9:4205-4206(1990).
RN [8]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 281-375.
RX PubMed=7507921; DOI=10.1128/jb.176.3.804-814.1994;
RA Pogliano K.J., Beckwith J.;
RT "Genetic and molecular characterization of the Escherichia coli secD operon
RT and its products.";
RL J. Bacteriol. 176:804-814(1994).
RN [9]
RP MUTAGENESIS OF SER-90.
RX PubMed=8003468; DOI=10.1021/bi00189a004;
RA Reuter K., Chong S., Ullrich F., Kersten H., Garcia G.A.;
RT "Serine 90 is required for enzymic activity by tRNA-guanine
RT transglycosylase from Escherichia coli.";
RL Biochemistry 33:7041-7046(1994).
RN [10]
RP MUTAGENESIS OF ZINC LIGANDS.
RX PubMed=7893665; DOI=10.1021/bi00011a026;
RA Chong S., Curnow A.W., Huston T.J., Garcia G.A.;
RT "tRNA-guanine transglycosylase from Escherichia coli is a zinc
RT metalloprotein. Site-directed mutagenesis studies to identify the zinc
RT ligands.";
RL Biochemistry 34:3694-3701(1995).
RN [11]
RP ACTIVE SITE CYS-265.
RX PubMed=9055203; DOI=10.1023/a:1026334726357;
RA Garcia G.A., Chong S.;
RT "Cysteine 265 is in the active site of, but is not essential for catalysis
RT by tRNA-guanine transglycosylase (TGT) from Escherichia coli.";
RL J. Protein Chem. 16:11-17(1997).
RN [12]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND ACTIVE
RP SITE.
RX PubMed=11714265; DOI=10.1021/bi0110589;
RA Kittendorf J.D., Barcomb L.M., Nonekowski S.T., Garcia G.A.;
RT "tRNA-guanine transglycosylase from Escherichia coli: molecular mechanism
RT and role of aspartate 89.";
RL Biochemistry 40:14123-14133(2001).
RN [13]
RP FUNCTION, CATALYTIC ACTIVITY, AND ACTIVE SITE.
RX PubMed=12909636; DOI=10.1074/jbc.m304323200;
RA Kittendorf J.D., Sgraja T., Reuter K., Klebe G., Garcia G.A.;
RT "An essential role for aspartate 264 in catalysis by tRNA-guanine
RT transglycosylase from Escherichia coli.";
RL J. Biol. Chem. 278:42369-42376(2003).
CC -!- FUNCTION: Catalyzes the base-exchange of a guanine (G) residue with the
CC queuine precursor 7-aminomethyl-7-deazaguanine (PreQ1) at position 34
CC (anticodon wobble position) in tRNAs with GU(N) anticodons (tRNA-Asp,
CC -Asn, -His and -Tyr). Catalysis occurs through a double-displacement
CC mechanism. The nucleophile active site attacks the C1' of nucleotide 34
CC to detach the guanine base from the RNA, forming a covalent enzyme-RNA
CC intermediate. The proton acceptor active site deprotonates the incoming
CC PreQ1, allowing a nucleophilic attack on the C1' of the ribose to form
CC the product. After dissociation, two additional enzymatic reactions on
CC the tRNA convert PreQ1 to queuine (Q), resulting in the hypermodified
CC nucleoside queuosine (7-(((4,5-cis-dihydroxy-2-cyclopenten-1-
CC yl)amino)methyl)-7-deazaguanosine). {ECO:0000255|HAMAP-Rule:MF_00168,
CC ECO:0000269|PubMed:11714265, ECO:0000269|PubMed:12909636}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=7-aminomethyl-7-carbaguanine + guanosine(34) in tRNA = 7-
CC aminomethyl-7-carbaguanosine(34) in tRNA + guanine;
CC Xref=Rhea:RHEA:24104, Rhea:RHEA-COMP:10341, Rhea:RHEA-COMP:10342,
CC ChEBI:CHEBI:16235, ChEBI:CHEBI:58703, ChEBI:CHEBI:74269,
CC ChEBI:CHEBI:82833; EC=2.4.2.29; Evidence={ECO:0000255|HAMAP-
CC Rule:MF_00168, ECO:0000269|PubMed:11714265,
CC ECO:0000269|PubMed:12909636};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00168,
CC ECO:0000269|PubMed:7893665};
CC Note=Binds 1 zinc ion per subunit. {ECO:0000255|HAMAP-Rule:MF_00168,
CC ECO:0000269|PubMed:7893665};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.12 uM for tRNA(Tyr) {ECO:0000269|PubMed:11714265};
CC KM=0.1 uM for guanine {ECO:0000269|PubMed:11714265};
CC Note=kcat is 0.00121 sec(-1) with tRNA(Tyr) and guanine as
CC substrates. {ECO:0000269|PubMed:11714265};
CC -!- PATHWAY: tRNA modification; tRNA-queuosine biosynthesis.
CC {ECO:0000255|HAMAP-Rule:MF_00168, ECO:0000305|PubMed:1706703}.
CC -!- SUBUNIT: Homodimer. Within each dimer, one monomer is responsible for
CC RNA recognition and catalysis, while the other monomer binds to the
CC replacement base PreQ1. {ECO:0000255|HAMAP-Rule:MF_00168}.
CC -!- SIMILARITY: Belongs to the queuine tRNA-ribosyltransferase family.
CC {ECO:0000255|HAMAP-Rule:MF_00168}.
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DR EMBL; M63939; AAA24667.1; -; Genomic_DNA.
DR EMBL; U00096; AAC73509.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE76186.1; -; Genomic_DNA.
DR EMBL; U82664; AAB40162.1; -; Genomic_DNA.
DR EMBL; X56175; CAA39631.1; ALT_SEQ; Genomic_DNA.
DR EMBL; X56175; CAA39632.1; ALT_SEQ; Genomic_DNA.
DR EMBL; S68715; AAC60467.2; -; Genomic_DNA.
DR PIR; C38530; C38530.
DR RefSeq; NP_414940.1; NC_000913.3.
DR RefSeq; WP_000667319.1; NZ_STEB01000007.1.
DR AlphaFoldDB; P0A847; -.
DR SMR; P0A847; -.
DR BioGRID; 4263531; 21.
DR DIP; DIP-36020N; -.
DR IntAct; P0A847; 39.
DR STRING; 511145.b0406; -.
DR jPOST; P0A847; -.
DR PaxDb; P0A847; -.
DR PRIDE; P0A847; -.
DR EnsemblBacteria; AAC73509; AAC73509; b0406.
DR EnsemblBacteria; BAE76186; BAE76186; BAE76186.
DR GeneID; 66671295; -.
DR GeneID; 949130; -.
DR KEGG; ecj:JW0396; -.
DR KEGG; eco:b0406; -.
DR PATRIC; fig|1411691.4.peg.1871; -.
DR EchoBASE; EB0989; -.
DR eggNOG; COG0343; Bacteria.
DR HOGENOM; CLU_022060_0_1_6; -.
DR InParanoid; P0A847; -.
DR OMA; GIDLFDC; -.
DR PhylomeDB; P0A847; -.
DR BioCyc; EcoCyc:EG10996-MON; -.
DR BioCyc; MetaCyc:EG10996-MON; -.
DR BRENDA; 2.4.2.29; 2026.
DR SABIO-RK; P0A847; -.
DR UniPathway; UPA00392; -.
DR PRO; PR:P0A847; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IBA:GO_Central.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0008479; F:queuine tRNA-ribosyltransferase activity; IDA:EcoCyc.
DR GO; GO:0008270; F:zinc ion binding; IDA:EcoCyc.
DR GO; GO:0008616; P:queuosine biosynthetic process; IMP:EcoCyc.
DR GO; GO:0002099; P:tRNA wobble guanine modification; IMP:EcoCyc.
DR GO; GO:0101030; P:tRNA-guanine transglycosylation; IEA:InterPro.
DR Gene3D; 3.20.20.105; -; 1.
DR HAMAP; MF_00168; Q_tRNA_Tgt; 1.
DR InterPro; IPR004803; TGT.
DR InterPro; IPR036511; TGT-like_sf.
DR InterPro; IPR002616; tRNA_ribo_trans-like.
DR Pfam; PF01702; TGT; 1.
DR SUPFAM; SSF51713; SSF51713; 1.
DR TIGRFAMs; TIGR00430; Q_tRNA_tgt; 1.
DR TIGRFAMs; TIGR00449; tgt_general; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; Glycosyltransferase; Metal-binding;
KW Queuosine biosynthesis; Reference proteome; Transferase; tRNA processing;
KW Zinc.
FT CHAIN 1..375
FT /note="Queuine tRNA-ribosyltransferase"
FT /id="PRO_0000135471"
FT REGION 245..251
FT /note="RNA binding"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00168"
FT REGION 269..273
FT /note="RNA binding; important for wobble base 34
FT recognition"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00168"
FT ACT_SITE 89
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00168,
FT ECO:0000305|PubMed:11714265"
FT ACT_SITE 264
FT /note="Nucleophile"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00168,
FT ECO:0000305|PubMed:12909636"
FT BINDING 89..93
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00168"
FT BINDING 143
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00168"
FT BINDING 187
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00168"
FT BINDING 214
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00168"
FT BINDING 302
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00168,
FT ECO:0000305|PubMed:7893665"
FT BINDING 304
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00168,
FT ECO:0000305|PubMed:7893665"
FT BINDING 307
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00168,
FT ECO:0000305|PubMed:7893665"
FT BINDING 333
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00168"
FT MUTAGEN 28
FT /note="C->A: Slight loss of activity."
FT /evidence="ECO:0000269|PubMed:7893665"
FT MUTAGEN 89
FT /note="D->E: Reduces catalytic activity by 51%."
FT /evidence="ECO:0000269|PubMed:11714265"
FT MUTAGEN 90
FT /note="S->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:8003468"
FT MUTAGEN 90
FT /note="S->C: Loss of activity."
FT /evidence="ECO:0000269|PubMed:8003468"
FT MUTAGEN 90
FT /note="S->F: Loss of activity."
FT /evidence="ECO:0000269|PubMed:8003468"
FT MUTAGEN 145
FT /note="C->A: No loss of activity."
FT /evidence="ECO:0000269|PubMed:7893665"
FT MUTAGEN 232
FT /note="C->A: Slight loss of activity."
FT /evidence="ECO:0000269|PubMed:7893665"
FT MUTAGEN 264
FT /note="D->A,N,Q,K,H: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:12909636"
FT MUTAGEN 264
FT /note="D->E: Reduces catalytic activity by 88%."
FT /evidence="ECO:0000269|PubMed:12909636"
FT MUTAGEN 265
FT /note="C->A: Significant loss of activity."
FT /evidence="ECO:0000269|PubMed:7893665"
FT MUTAGEN 302
FT /note="C->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:7893665"
FT MUTAGEN 304
FT /note="C->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:7893665"
FT MUTAGEN 307
FT /note="C->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:7893665"
FT MUTAGEN 316
FT /note="H->A: Slight loss of activity."
FT /evidence="ECO:0000269|PubMed:7893665"
FT MUTAGEN 317
FT /note="H->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:7893665"
FT MUTAGEN 321
FT /note="C->A: No loss of activity."
FT /evidence="ECO:0000269|PubMed:7893665"
FT CONFLICT 54..55
FT /note="IL -> MV (in Ref. 1 and 2)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 375 AA; 42594 MW; 40271CA08D8A8820 CRC64;
MKFELDTTDG RARRGRLVFD RGVVETPCFM PVGTYGTVKG MTPEEVEATG AQIILGNTFH
LWLRPGQEIM KLHGDLHDFM QWKGPILTDS GGFQVFSLGD IRKITEQGVH FRNPINGDPI
FLDPEKSMEI QYDLGSDIVM IFDECTPYPA DWDYAKRSME MSLRWAKRSR ERFDSLGNKN
ALFGIIQGSV YEDLRDISVK GLVDIGFDGY AVGGLAVGEP KADMHRILEH VCPQIPADKP
RYLMGVGKPE DLVEGVRRGI DMFDCVMPTR NARNGHLFVT DGVVKIRNAK YKSDTGPLDP
ECDCYTCRNY SRAYLHHLDR CNEILGARLN TIHNLRYYQR LMAGLRKAIE EGKLESFVTD
FYQRQGREVP PLNVD
