TUT1_LEITA
ID TUT1_LEITA Reviewed; 1120 AA.
AC Q8WQX6;
DT 12-AUG-2020, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2002, sequence version 1.
DT 03-AUG-2022, entry version 52.
DE RecName: Full=Terminal uridylyltransferase 1 {ECO:0000305};
DE Short=TUTase 1 {ECO:0000303|PubMed:11893335};
DE EC=2.7.7.52 {ECO:0000269|PubMed:11893335, ECO:0000269|PubMed:15060068};
DE AltName: Full=3' terminal uridylyl transferase {ECO:0000303|PubMed:11893335};
DE Short=3' TUTase {ECO:0000303|PubMed:11893335};
DE AltName: Full=RNA editing terminal uridylyltransferase 1 {ECO:0000305};
DE Short=RET1 {ECO:0000305};
DE Short=RNA editing TUTase 1 {ECO:0000305};
DE Flags: Precursor;
GN Name=KRET1 {ECO:0000305};
GN ORFNames=LtaPh_1409400 {ECO:0000312|EMBL:GET87125.1};
OS Leishmania tarentolae (Sauroleishmania tarentolae).
OC Eukaryota; Discoba; Euglenozoa; Kinetoplastea; Metakinetoplastina;
OC Trypanosomatida; Trypanosomatidae; Leishmaniinae; Leishmania;
OC lizard Leishmania.
OX NCBI_TaxID=5689 {ECO:0000312|EMBL:AAK38333.1};
RN [1] {ECO:0000312|EMBL:AAK38333.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, COFACTOR,
RP SUBUNIT, INTERACTION WITH P45 AND P50 RNA LIGASES, AND SUBCELLULAR
RP LOCATION.
RX PubMed=11893335; DOI=10.1016/s0092-8674(02)00647-5;
RA Aphasizhev R., Sbicego S., Peris M., Jang S.H., Aphasizheva I.,
RA Simpson A.M., Rivlin A., Simpson L.;
RT "Trypanosome mitochondrial 3' terminal uridylyl transferase (TUTase): the
RT key enzyme in U-insertion/deletion RNA editing.";
RL Cell 108:637-648(2002).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=Parrot Tar II;
RA Goto Y., Yamagishi J.;
RT "Leishmania tarentolae CDS.";
RL Submitted (NOV-2019) to the EMBL/GenBank/DDBJ databases.
RN [3] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBUNIT, AND MUTAGENESIS OF PHE-328; SER-330; ASP-342; ASP-344;
RP ARG-390; LYS-395; 400-LEU--TYR-543; ASP-548; LYS-580; SER-598; TYR-599 AND
RP ASP-775.
RX PubMed=15060068; DOI=10.1074/jbc.m401234200;
RA Aphasizheva I., Aphasizhev R., Simpson L.;
RT "RNA-editing terminal uridylyl transferase 1: identification of functional
RT domains by mutational analysis.";
RL J. Biol. Chem. 279:24123-24130(2004).
CC -!- FUNCTION: Terminal uridylyltransferase which is involved in the post-
CC transcriptional editing of mitochondrial RNA, a process involving the
CC addition and deletion of uridine (U) nucleotides in the pre-mRNA.
CC Specifically, catalyzes the addition of Us to the 3'-hydroxyl group of
CC guided RNA (gRNA), with a preference for RNAs terminating in 6 Us, but
CC also can add Us to RNAs terminating in 6 adenines (A), 6 cytosines (C),
CC or 6 guanines (G) (PubMed:11893335, PubMed:15060068). Can mediate RNA-
CC independent UTP polymerization in vitro (PubMed:15060068). Can mediate
CC pyrophosphate-dependent degradation of synthetic RNA ending with U
CC residues in vitro (PubMed:15060068). {ECO:0000269|PubMed:11893335,
CC ECO:0000269|PubMed:15060068}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=RNA(n) + UTP = diphosphate + RNA(n)-3'-uridine ribonucleotide;
CC Xref=Rhea:RHEA:14785, Rhea:RHEA-COMP:14527, Rhea:RHEA-COMP:17348,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:46398, ChEBI:CHEBI:140395,
CC ChEBI:CHEBI:173116; EC=2.7.7.52;
CC Evidence={ECO:0000269|PubMed:11893335, ECO:0000269|PubMed:15060068};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000305|PubMed:11893335};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000305|PubMed:11893335};
CC Note=Binds 1 Mg(2+) or Mn(2+) per subunit (PubMed:11893335). The type
CC of divalent cation used by the enzyme affects the nucleotide
CC specificity; Mg(2+) induces predominantly uridine (U) incorporation
CC while Mn(2+) induces also substantial incorporation of both adenine (A)
CC and cytosine (C) (PubMed:11893335). {ECO:0000269|PubMed:11893335};
CC -!- ACTIVITY REGULATION: Zinc-binding is required for catalytic activity.
CC {ECO:0000269|PubMed:15060068}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC Note=KM is 45-65 uM for UTP (at 27 degrees Celsius)
CC (PubMed:15060068). KM is 18-30 uM for RNA (at 27 degrees Celsius)
CC (PubMed:15060068). {ECO:0000269|PubMed:15060068};
CC -!- SUBUNIT: Homotetramer (PubMed:11893335, PubMed:15060068). Part of a
CC 700kDa complex (PubMed:11893335). Interacts with p45 and p50 RNA
CC ligases (PubMed:11893335). {ECO:0000269|PubMed:11893335,
CC ECO:0000269|PubMed:15060068}.
CC -!- SUBCELLULAR LOCATION: Mitochondrion {ECO:0000269|PubMed:11893335}.
CC -!- SIMILARITY: Belongs to the DNA polymerase type-B-like family.
CC {ECO:0000305}.
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DR EMBL; AY029069; AAK38333.1; -; Genomic_DNA.
DR EMBL; BLBS01000018; GET87125.1; -; Genomic_DNA.
DR AlphaFoldDB; Q8WQX6; -.
DR SMR; Q8WQX6; -.
DR VEuPathDB; TriTrypDB:LtaPh_1409400; -.
DR PhylomeDB; Q8WQX6; -.
DR GO; GO:0005739; C:mitochondrion; IDA:UniProtKB.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0003723; F:RNA binding; IEA:UniProtKB-KW.
DR GO; GO:0050265; F:RNA uridylyltransferase activity; IDA:UniProtKB.
DR GO; GO:0006397; P:mRNA processing; IEA:UniProtKB-KW.
DR GO; GO:0051260; P:protein homooligomerization; IDA:UniProtKB.
DR GO; GO:0071076; P:RNA 3' uridylation; IDA:UniProtKB.
DR Gene3D; 3.30.460.10; -; 1.
DR InterPro; IPR043519; NT_sf.
DR InterPro; IPR002058; PAP_assoc.
DR Pfam; PF03828; PAP_assoc; 1.
DR SUPFAM; SSF81301; SSF81301; 1.
DR PROSITE; PS50157; ZINC_FINGER_C2H2_2; 1.
PE 1: Evidence at protein level;
KW Magnesium; Manganese; Metal-binding; Mitochondrion; mRNA processing;
KW Nucleotide-binding; Nucleotidyltransferase; RNA-binding; Transferase;
KW Transit peptide; Zinc; Zinc-finger.
FT TRANSIT 1..?
FT /note="Mitochondrion"
FT /evidence="ECO:0000305"
FT CHAIN ?..1120
FT /note="Terminal uridylyltransferase 1"
FT /id="PRO_0000450679"
FT DOMAIN 659..697
FT /note="PAP-associated"
FT /evidence="ECO:0000255"
FT ZN_FING 222..253
FT /note="C2H2-type; atypical"
FT /evidence="ECO:0000250|UniProtKB:Q8WQX5"
FT REGION 1..156
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 196..221
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 750..1120
FT /note="Important for catalytic activity and RNA binding"
FT /evidence="ECO:0000269|PubMed:15060068"
FT REGION 800..900
FT /note="Involved in oligomerization"
FT /evidence="ECO:0000269|PubMed:15060068"
FT REGION 1047..1076
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT MOTIF 773..782
FT /note="Nucleotide recognition motif (NRM)"
FT /evidence="ECO:0000250|UniProtKB:Q8WQX5"
FT COMPBIAS 1..56
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 87..101
FT /note="Basic and acidic residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 102..128
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 205..221
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 1047..1068
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT BINDING 227
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q8WQX5"
FT BINDING 230
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q8WQX5"
FT BINDING 244
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q8WQX5"
FT BINDING 249
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q8WQX5"
FT BINDING 330
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000250|UniProtKB:Q381M1"
FT BINDING 341..344
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000250|UniProtKB:Q381M1"
FT BINDING 342
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000250|UniProtKB:Q86MV5"
FT BINDING 344
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000250|UniProtKB:Q86MV5"
FT BINDING 390
FT /ligand="RNA"
FT /ligand_id="ChEBI:CHEBI:33697"
FT /evidence="ECO:0000250|UniProtKB:Q381M1"
FT BINDING 548
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000305|PubMed:15060068"
FT BINDING 555..559
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000250|UniProtKB:Q8WQX5"
FT BINDING 580
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000250|UniProtKB:Q8WQX5"
FT BINDING 584
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000250|UniProtKB:Q8WQX5"
FT BINDING 598..599
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000250|UniProtKB:Q8WQX5"
FT SITE 548
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000250|UniProtKB:Q86MV5"
FT MUTAGEN 328
FT /note="F->G: 3-fold reduction in affinity for RNA. Does not
FT affect affinity for UTP."
FT /evidence="ECO:0000269|PubMed:15060068"
FT MUTAGEN 330
FT /note="S->A: 50-fold reduction in affinity for RNA and 7-
FT fold increase in affinity for UTP."
FT /evidence="ECO:0000269|PubMed:15060068"
FT MUTAGEN 342
FT /note="D->A: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:15060068"
FT MUTAGEN 344
FT /note="D->A: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:15060068"
FT MUTAGEN 390
FT /note="R->F: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:15060068"
FT MUTAGEN 395
FT /note="K->D: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:15060068"
FT MUTAGEN 400..543
FT /note="Missing: Loss of catalytic activity. Does not affect
FT tetramerization."
FT /evidence="ECO:0000269|PubMed:15060068"
FT MUTAGEN 548
FT /note="D->A: Loss of catalytic activity. Does not affect
FT gRNA binding."
FT /evidence="ECO:0000269|PubMed:15060068"
FT MUTAGEN 580
FT /note="K->A: 6-fold reduction in affinity for UTP. Does not
FT affect affinity for RNA."
FT /evidence="ECO:0000269|PubMed:15060068"
FT MUTAGEN 598
FT /note="S->A: Does not affect affinity for RNA and UTP."
FT /evidence="ECO:0000269|PubMed:15060068"
FT MUTAGEN 599
FT /note="Y->F: Does not affect affinity for RNA and UTP."
FT /evidence="ECO:0000269|PubMed:15060068"
FT MUTAGEN 775
FT /note="D->A: 25-fold reduction in affinity for RNA. 2-fold
FT increase in affinity for UTP."
FT /evidence="ECO:0000269|PubMed:15060068"
SQ SEQUENCE 1120 AA; 120930 MW; EA45A807ECB0A328 CRC64;
MSKYSLLFNQ GTKDGTNASS GSEANSANIT SSSAPASSTN TSSPTSSESA VVSPPASTSP
RRRLIHRRHG SAGAAEVAPL SLPKRPQQPN EEKHENFISD SVHHCSNRGA SGSELKALTT
SGSETVMSAS PDIAFEAPSP PTASASPPLE STSAVESDGD VVIDDMMRYQ EGDSGGSRSA
TSAAAAGRAV STNDAAALIN GDPGPLSSAV SSSSSGSPHT PPRLFTCDMC LQYVSTSYEA
LEQHALDQHG DALADYTRLR SVAEKLVPVW DEVLKRKASV VQQWGKRIFA VAVQRDAGAE
KMAEAHRARA QLELVVQRWH PRAKVFIFGS SVAFGVWDGI SDIDFTVVDV DELEAGTWPP
SEKNAVRSIT ELLRRAGFSF INLEPISHAR VPIIKHHASL PIRLTDEQRH RLYEEARQSA
AAVDLVAAES LASSSPSSAQ ETTDEKGLTQ LEAELIIARS VRYSLNLPAG PPDSAILEAS
IRLAVGSAAV QQVWWNRTRD MCCMTFDTTT NAVKASTCPL HFMSAGMRAR VQPLHEECRP
ELYGMDFDLS FRAFGIRNSH LLRRYLLSHP CARPGALVLK DWSKTSGVNN SVNGYLTSYA
INIMWIYYLV HRGVIRYVCP ARDIPASLRC NVDADPQYAA MVDPTWTPEE RAAMEAQAGE
LLLGFFYYYA FEFDWVNHVV SLNRPGITTK ASLGWDVEDV AQTGSPAPHF GVAGSQHQYN
LAGAEGQQGD LHSGTSLSAP QTRPLTGYDG MMASSASAAA RRSRATTRYS FCIEDPYEEN
LNLGRHMGVT KTLRVQTELY RGLLSLLKDD PQHCCVFAGS TNSSGSTDSN GAMASGAAEP
AMVAARTPSE PTELPVRVLY KLMAISTREL AIARRRYSAT VTAGTEFPGA LLSDLEAAFL
AQAPTEWKLA RQVWNKHQLL HRLGLKLHAR EYVLPRREVG VRRLAAKAPP GVVPASAPEP
TFTAEEVAAA AAESGQAPFS AEHAPTSSEE VTQMNRAFLG AFPARRLPED LMLAMTKGYS
CLTPSWVAWS KPWAALSAWW TDRLHSPSTT TQGEDPLASG TCEQGGVSPS LPTGAPHHIS
AVPEKSAGAM HQTRTQLRRH VVAEIASTPA ARRVLRLLFR
