TUT2_TRYBB
ID TUT2_TRYBB Reviewed; 487 AA.
AC Q86MV5;
DT 22-APR-2020, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-2003, sequence version 1.
DT 03-AUG-2022, entry version 57.
DE RecName: Full=Terminal uridylyltransferase 2 {ECO:0000305};
DE Short=TUTase 2 {ECO:0000305};
DE EC=2.7.7.52 {ECO:0000269|PubMed:12820966, ECO:0000269|PubMed:16281058, ECO:0000269|PubMed:19465686, ECO:0000269|PubMed:20362585};
DE AltName: Full=Mitochondrial protein 57 {ECO:0000303|PubMed:12649499};
DE Short=TbMP57 {ECO:0000303|PubMed:12649499};
DE AltName: Full=RNA editing 3' terminal uridylyltransferase 2 {ECO:0000303|PubMed:12820966};
DE Short=RET2 {ECO:0000303|PubMed:12820966};
DE Short=RNA editing TUTase 2 {ECO:0000303|PubMed:12820966};
DE AltName: Full=RNA editing complex protein MP57 {ECO:0000303|PubMed:12820966};
DE Flags: Precursor;
GN Name=KRET2 {ECO:0000305}; Synonyms=MP57 {ECO:0000312|EMBL:AAO63567.1};
OS Trypanosoma brucei brucei.
OC Eukaryota; Discoba; Euglenozoa; Kinetoplastea; Metakinetoplastina;
OC Trypanosomatida; Trypanosomatidae; Trypanosoma.
OX NCBI_TaxID=5702;
RN [1] {ECO:0000312|EMBL:AAO63567.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP IDENTIFICATION IN THE RECC COMPLEX, INTERACTION WITH MP81, AND SUBCELLULAR
RP LOCATION.
RC STRAIN=IsTar I.7 {ECO:0000312|EMBL:AAO63567.1};
RX PubMed=12820966; DOI=10.1016/s1097-2765(03)00185-0;
RA Ernst N.L., Panicucci B., Igo R.P., Panigrahi A.K., Salavati R., Stuart K.;
RT "TbMP57 is a 3' terminal uridylyl transferase (TUTase) of the Trypanosoma
RT brucei editosome.";
RL Mol. Cell 11:1525-1536(2003).
RN [2] {ECO:0000312|EMBL:AAO63567.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], IDENTIFICATION IN THE RECC COMPLEX, AND
RP SUBCELLULAR LOCATION.
RC STRAIN=IsTar I.7 {ECO:0000312|EMBL:AAO63567.1};
RX PubMed=12649499; DOI=10.1261/rna.2194603;
RA Panigrahi A.K., Schnaufer A., Ernst N.L., Wang B., Carmean N., Salavati R.,
RA Stuart K.;
RT "Identification of novel components of Trypanosoma brucei editosomes.";
RL RNA 9:484-492(2003).
RN [3] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, IDENTIFICATION
RP IN THE RECC COMPLEX, INTERACTION WITH MP81, AND SUBCELLULAR LOCATION.
RX PubMed=19465686; DOI=10.1261/rna.1538809;
RA Aphasizheva I., Ringpis G.E., Weng J., Gershon P.D., Lathrop R.H.,
RA Aphasizhev R.;
RT "Novel TUTase associates with an editosome-like complex in mitochondria of
RT Trypanosoma brucei.";
RL RNA 15:1322-1337(2009).
RN [4] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, IDENTIFICATION
RP IN THE RECC COMPLEX, INTERACTION WITH MP81, DOMAIN, AND DISRUPTION
RP PHENOTYPE.
RX PubMed=20362585; DOI=10.1016/j.jmb.2010.03.050;
RA Ringpis G.E., Aphasizheva I., Wang X., Huang L., Lathrop R.H.,
RA Hatfield G.W., Aphasizhev R.;
RT "Mechanism of U insertion RNA editing in trypanosome mitochondria: the
RT bimodal TUTase activity of the core complex.";
RL J. Mol. Biol. 399:680-695(2010).
RN [5] {ECO:0007744|PDB:2B4V, ECO:0007744|PDB:2B51, ECO:0007744|PDB:2B56}
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) OF 20-487 OF MUTANT TRP-94; TRP-122
RP AND TRP-207 IN COMPLEX WITH MAGNESIUM; MANGANESE AND UTP, FUNCTION,
RP CATALYTIC ACTIVITY, COFACTOR, AND MUTAGENESIS OF TRP-94; TRP-122 AND
RP TRP-207.
RX PubMed=16281058; DOI=10.1038/sj.emboj.7600861;
RA Deng J., Ernst N.L., Turley S., Stuart K.D., Hol W.G.;
RT "Structural basis for UTP specificity of RNA editing TUTases from
RT Trypanosoma brucei.";
RL EMBO J. 24:4007-4017(2005).
CC -!- FUNCTION: Terminal uridylyltransferase which, as part of the
CC mitochondrial RNA editing core complex (RECC), 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 (PubMed:12820966, PubMed:19465686, PubMed:20362585). Specifically,
CC catalyzes the addition of one U to single-stranded RNA with a
CC preference for a 3'-terminal A or G and adds the number of Us specified
CC by a guide RNA (gRNA) to precleaved double-stranded RNA editing
CC substrates (PubMed:12820966, PubMed:19465686, PubMed:20362585,
CC PubMed:16281058). Essential for the survival of the bloodstream form
CC (PubMed:16281058). {ECO:0000269|PubMed:12820966,
CC ECO:0000269|PubMed:16281058, ECO:0000269|PubMed:19465686,
CC ECO:0000269|PubMed:20362585}.
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:12820966, ECO:0000269|PubMed:16281058,
CC ECO:0000269|PubMed:19465686, ECO:0000269|PubMed:20362585};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000305|PubMed:16281058};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000305|PubMed:16281058};
CC Note=Binds 1 Mg(2+) or Mn(2+) per subunit.
CC {ECO:0000269|PubMed:16281058};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.2 uM for UTP (with 6(U) single-stranded RNA as substrate)
CC {ECO:0000269|PubMed:19465686};
CC KM=3.4 uM for UTP (with double-stranded RNA as substrate)
CC {ECO:0000269|PubMed:19465686};
CC KM=2.5 uM for UTP (with RNA with no U in 3' position as substrate)
CC {ECO:0000269|PubMed:19465686};
CC KM=12.6 uM for UTP (with double-stranded RNA with one U in 3'
CC position as substrate) {ECO:0000269|PubMed:19465686};
CC Note=kcat is 0.0007 min(-1) with UTP and 6(U) single-stranded RNA as
CC substrates (PubMed:19465686). kcat is 0.002 min(-1) with UTP and
CC double-stranded RNA as substrates (PubMed:19465686). kcat is 0.02
CC min(-1) with UTP and with RNA with no U in 3' position as substrates
CC (PubMed:20362585). kcat is 0.2 min(-1) with UTP and double-stranded
CC RNA with one U in 3' position as substrates (PubMed:20362585).
CC {ECO:0000269|PubMed:19465686, ECO:0000269|PubMed:20362585};
CC -!- SUBUNIT: Component of the mitochondrial RNA editing core complex
CC (RECC), also known as the editosome complex (PubMed:12820966,
CC PubMed:12649499, PubMed:19465686, PubMed:20362585). Interacts (via
CC middle domain) with MP81; the interaction enhances MP57 catalytic
CC activity and probably recruits MP57 to the RECC complex
CC (PubMed:12820966, PubMed:20362585). {ECO:0000269|PubMed:12649499,
CC ECO:0000269|PubMed:12820966, ECO:0000269|PubMed:19465686,
CC ECO:0000269|PubMed:20362585}.
CC -!- SUBCELLULAR LOCATION: Mitochondrion {ECO:0000269|PubMed:12649499,
CC ECO:0000269|PubMed:12820966, ECO:0000269|PubMed:19465686}.
CC -!- DISRUPTION PHENOTYPE: RNAi-mediated knockdown in the procyclic form
CC causes growth arrest. {ECO:0000269|PubMed:20362585}.
CC -!- SIMILARITY: Belongs to the DNA polymerase type-B-like 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; AY228173; AAO63567.1; -; Genomic_DNA.
DR PDB; 2B4V; X-ray; 1.80 A; A=20-487.
DR PDB; 2B51; X-ray; 2.05 A; A=20-487.
DR PDB; 2B56; X-ray; 1.97 A; A=20-487.
DR PDBsum; 2B4V; -.
DR PDBsum; 2B51; -.
DR PDBsum; 2B56; -.
DR AlphaFoldDB; Q86MV5; -.
DR SMR; Q86MV5; -.
DR EvolutionaryTrace; Q86MV5; -.
DR GO; GO:0031019; C:mitochondrial mRNA editing complex; IDA:UniProtKB.
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:0071076; P:RNA 3' uridylation; IDA:UniProtKB.
DR InterPro; IPR043519; NT_sf.
DR InterPro; IPR002058; PAP_assoc.
DR InterPro; IPR041060; Ret2_MD.
DR Pfam; PF03828; PAP_assoc; 1.
DR Pfam; PF18528; Ret2_MD; 1.
DR SUPFAM; SSF81301; SSF81301; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Magnesium; Manganese; Metal-binding; Mitochondrion;
KW mRNA processing; Nucleotide-binding; Nucleotidyltransferase; RNA-binding;
KW Transferase; Transit peptide.
FT TRANSIT 1..24
FT /note="Mitochondrion"
FT /evidence="ECO:0000255"
FT CHAIN 25..487
FT /note="Terminal uridylyltransferase 2"
FT /id="PRO_0000449385"
FT DOMAIN 366..425
FT /note="PAP-associated"
FT /evidence="ECO:0000255"
FT REGION 151..264
FT /note="Middle domain (MD); important for the incorporation
FT into the RECC complex, for catalytic activity and possibly
FT RNA binding"
FT /evidence="ECO:0000269|PubMed:20362585"
FT BINDING 85
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000269|PubMed:16281058,
FT ECO:0007744|PDB:2B51, ECO:0007744|PDB:2B56"
FT BINDING 96..99
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000269|PubMed:16281058,
FT ECO:0007744|PDB:2B51, ECO:0007744|PDB:2B56"
FT BINDING 97
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:16281058"
FT BINDING 99
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:16281058"
FT BINDING 210
FT /ligand="RNA"
FT /ligand_id="ChEBI:CHEBI:33697"
FT /evidence="ECO:0000250|UniProtKB:Q381M1"
FT BINDING 277..278
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000269|PubMed:16281058,
FT ECO:0007744|PDB:2B51, ECO:0007744|PDB:2B56"
FT BINDING 300
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000269|PubMed:16281058,
FT ECO:0007744|PDB:2B51, ECO:0007744|PDB:2B56"
FT BINDING 304
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000269|PubMed:16281058,
FT ECO:0007744|PDB:2B51, ECO:0007744|PDB:2B56"
FT BINDING 318..319
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000269|PubMed:16281058,
FT ECO:0007744|PDB:2B51, ECO:0007744|PDB:2B56"
FT SITE 267
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000305|PubMed:16281058"
FT MUTAGEN 94
FT /note="W->R: No effect on catalytic activity; when
FT associated with R-122 and R-207."
FT /evidence="ECO:0000269|PubMed:16281058"
FT MUTAGEN 122
FT /note="W->R: No effect on catalytic activity; when
FT associated with R-94 and R-207."
FT /evidence="ECO:0000269|PubMed:16281058"
FT MUTAGEN 207
FT /note="W->R: No effect on catalytic activity; when
FT associated with R-94 and R-122."
FT /evidence="ECO:0000269|PubMed:16281058"
FT HELIX 33..50
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 54..74
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 79..83
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 85..89
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 98..103
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 105..108
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 111..126
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 131..134
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 137..140
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 142..145
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 147..150
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 155..163
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 168..174
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 176..183
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 188..198
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 200..202
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 203..210
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 212..223
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 224..231
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 237..240
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 242..247
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 248..253
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 260..262
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 266..270
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 273..287
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 292..304
FT /evidence="ECO:0007829|PDB:2B4V"
FT TURN 307..309
FT /evidence="ECO:0007829|PDB:2B56"
FT STRAND 311..314
FT /evidence="ECO:0007829|PDB:2B56"
FT HELIX 318..331
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 340..342
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 346..348
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 363..379
FT /evidence="ECO:0007829|PDB:2B4V"
FT TURN 383..385
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 386..388
FT /evidence="ECO:0007829|PDB:2B4V"
FT STRAND 390..394
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 398..401
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 406..408
FT /evidence="ECO:0007829|PDB:2B4V"
FT TURN 424..429
FT /evidence="ECO:0007829|PDB:2B4V"
FT TURN 433..436
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 439..458
FT /evidence="ECO:0007829|PDB:2B4V"
FT TURN 461..463
FT /evidence="ECO:0007829|PDB:2B4V"
FT HELIX 468..470
FT /evidence="ECO:0007829|PDB:2B4V"
SQ SEQUENCE 487 AA; 56560 MW; 3B57EB6812D0619A CRC64;
MLMHTAPWLH MRLSRLFRQS PLSLPSTKLN PSPDHYAVWG KAIMAENNRR VGPEHMFRTA
IRAQQQLQGL ADKWTPDAKV YCCGSMVTYG QMEWGSDLDL ACMFDDPYPS HEVQAKRTDK
LWTVIKRYVP HYLRNNLLGL TEARTPVVKL RFANDEKVAR ARYTPLSEEE DRKARTALLD
VRNQCVGDND VEYIAEKMGR DNVEGIWVDR TTYGCRIAIQ CTSKEQMIEA IGFFPDGKIM
TRGMREDYTR DVLDVRFVPE MFMYRWDISF VGYGVKNSYL IRHYLHNGPV AARHTAMAVK
AWGKATNVGA GSGAMLTSYA VTVMFIYYLL VTRQVLWVDP WSLPHPAHLP RYPDFSPLYD
CDPTELGRLL HGFFIFYAHH FDYEREVVSL NRNRRSYRSD IGWNFPQNKK GTFSYNFCIE
DPYEDVGTGG LNLVRHLHPA KFQLVKQEFL RAAQCMERFL PTNAPEKSIL GVKRADLRHF
ERDRDRE
