TUT7_TRYBB
ID TUT7_TRYBB Reviewed; 406 AA.
AC C7AJA4;
DT 22-APR-2020, integrated into UniProtKB/Swiss-Prot.
DT 22-APR-2020, sequence version 2.
DT 03-AUG-2022, entry version 20.
DE RecName: Full=Terminal uridylyltransferase 7 {ECO:0000305};
DE Short=TUTase 7 {ECO:0000305};
DE EC=2.7.7.52 {ECO:0000269|PubMed:19465686, ECO:0000269|PubMed:20403364};
DE AltName: Full=Mitochondrial editosome-like complex associated TUTase {ECO:0000303|PubMed:19465686};
DE Short=TbMEAT1 {ECO:0000303|PubMed:20403364};
DE Flags: Precursor;
GN Name=MEAT1 {ECO:0000303|PubMed:19465686};
OS Trypanosoma brucei brucei.
OC Eukaryota; Discoba; Euglenozoa; Kinetoplastea; Metakinetoplastina;
OC Trypanosomatida; Trypanosomatidae; Trypanosoma.
OX NCBI_TaxID=5702;
RN [1] {ECO:0000312|EMBL:ACT83521.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, COFACTOR,
RP BIOPHYSICOCHEMICAL PROPERTIES, IDENTIFICATION IN THE RECC-LIKE COMPLEX,
RP INTERACTION WITH REL1, SUBCELLULAR LOCATION, DISRUPTION PHENOTYPE, AND
RP MUTAGENESIS OF ASP-67.
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 [2] {ECO:0007744|PDB:3HIY, ECO:0007744|PDB:3HJ1, ECO:0007744|PDB:3HJ4}
RP X-RAY CRYSTALLOGRAPHY (1.56 ANGSTROMS) OF 3-385 IN COMPLEX WITH UTP AND
RP MAGNESIUM, FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND MUTAGENESIS OF
RP 2-ASN--HIS-25; ASP-67; ASN-141; ASN-181; PHE-183; ASP-335; ASN-338;
RP CYS-342; ARG-344 AND ARG-345.
RX PubMed=20403364; DOI=10.1016/j.jmb.2010.04.021;
RA Stagno J., Aphasizheva I., Bruystens J., Luecke H., Aphasizhev R.;
RT "Structure of the mitochondrial editosome-like complex associated TUTase 1
RT reveals divergent mechanisms of UTP selection and domain organization.";
RL J. Mol. Biol. 399:464-475(2010).
CC -!- FUNCTION: Terminal uridylyltransferase which, as part of the
CC mitochondrial RNA editing core-like complex (RECC-like), is involved in
CC the post-transcriptional editing of mitochondrial RNA, a process
CC involving the addition and deletion of uridine (U) nucleotides in the
CC pre-mRNA (PubMed:19465686). Specifically, catalyzes the addition of U
CC to single-stranded RNA with a preference for a 3'-terminal U and adds
CC the number of Us specified by a guide RNA (gRNA) to precleaved double-
CC stranded RNA editing substrates (PubMed:19465686, PubMed:20403364).
CC Essential for insect and bloodstream developmental forms viability
CC (PubMed:19465686). {ECO:0000269|PubMed:19465686,
CC ECO:0000269|PubMed:20403364}.
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:19465686, ECO:0000269|PubMed:20403364};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:19465686};
CC Note=Binds 1 Mg(2+) per subunit. {ECO:0000269|PubMed:20403364};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.6 uM for UTP (with 6(U) single-stranded RNA as substrate)
CC {ECO:0000269|PubMed:19465686};
CC KM=1 uM for UTP (with double-stranded RNA as substrate)
CC {ECO:0000269|PubMed:19465686};
CC Note=kcat is 0.18 min(-1) with UTP and 6(U) single-stranded RNA as
CC substrates (PubMed:19465686). kcat is 0.04 min(-1) with UTP and
CC double-stranded RNA as substrates (PubMed:19465686).
CC {ECO:0000269|PubMed:19465686};
CC -!- SUBUNIT: Component of the mitochondrial RNA editing core complex-like
CC (RECC-like), also known as the editosome-like complex; only a small
CC proportion of MEAT1 associates with the complex (PubMed:19465686).
CC Interacts with RNA-editing ligase REL1 (PubMed:19465686).
CC {ECO:0000269|PubMed:19465686}.
CC -!- SUBCELLULAR LOCATION: Mitochondrion matrix
CC {ECO:0000269|PubMed:19465686}.
CC -!- DISRUPTION PHENOTYPE: RNAi-mediated knockdown in the procyclic form
CC causes growth arrest (PubMed:19465686). Moderately increases the
CC abundance of mitochondrial RNAs without causing defects in RNA editing
CC (PubMed:19465686). {ECO:0000269|PubMed:19465686}.
CC -!- SIMILARITY: Belongs to the DNA polymerase type-B-like family.
CC {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=ACT83521.1; Type=Miscellaneous discrepancy; Note=The submitted sequence corresponds to a mutated form and contains an Ala residue at position 67 instead of an Asp residue.; Evidence={ECO:0000305};
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DR EMBL; FJ561337; ACT83521.1; ALT_SEQ; Genomic_DNA.
DR PDB; 3HIY; X-ray; 2.30 A; A/B=3-385.
DR PDB; 3HJ1; X-ray; 1.95 A; A/B=1-385.
DR PDB; 3HJ4; X-ray; 1.56 A; A/B=3-385.
DR PDBsum; 3HIY; -.
DR PDBsum; 3HJ1; -.
DR PDBsum; 3HJ4; -.
DR AlphaFoldDB; C7AJA4; -.
DR SMR; C7AJA4; -.
DR OMA; KILCAIR; -.
DR BRENDA; 2.7.7.52; 6519.
DR GO; GO:0005759; C:mitochondrial matrix; IEA:UniProtKB-SubCell.
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: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 Gene3D; 3.30.460.10; -; 1.
DR InterPro; IPR043519; NT_sf.
DR SUPFAM; SSF81301; SSF81301; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Magnesium; Metal-binding; Mitochondrion; mRNA processing;
KW Nucleotide-binding; Nucleotidyltransferase; Transferase; Transit peptide.
FT TRANSIT 1..15
FT /note="Mitochondrion"
FT /evidence="ECO:0000255"
FT CHAIN 16..406
FT /note="Terminal uridylyltransferase 7"
FT /evidence="ECO:0000255"
FT /id="PRO_0000449794"
FT BINDING 54
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000269|PubMed:20403364,
FT ECO:0007744|PDB:3HIY, ECO:0007744|PDB:3HJ1"
FT BINDING 64..65
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000269|PubMed:20403364,
FT ECO:0007744|PDB:3HIY"
FT BINDING 65
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:20403364,
FT ECO:0007744|PDB:3HIY"
FT BINDING 67
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:20403364,
FT ECO:0007744|PDB:3HIY"
FT BINDING 138..142
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000269|PubMed:20403364,
FT ECO:0007744|PDB:3HIY, ECO:0007744|PDB:3HJ1"
FT BINDING 164
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000269|PubMed:20403364,
FT ECO:0007744|PDB:3HIY, ECO:0007744|PDB:3HJ1"
FT BINDING 168
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000269|PubMed:20403364,
FT ECO:0007744|PDB:3HIY, ECO:0007744|PDB:3HJ1"
FT BINDING 181..183
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000269|PubMed:20403364,
FT ECO:0007744|PDB:3HIY, ECO:0007744|PDB:3HJ1"
FT MUTAGEN 2..25
FT /note="Missing: No effect on catalytic activity. However,
FT shows a moderate lengthening in the extension of the
FT product at longer incubation time. Complete loss of
FT catalytic activity; when associated with A-67; A-181; D-
FT 181; F-183; A-335 and A-345. Partial loss of catalytic
FT activity; when associated with A-141; A-338 and R-344. No
FT loss of catalytic activity; when associated with A-344."
FT /evidence="ECO:0000269|PubMed:20403364"
FT MUTAGEN 67
FT /note="D->A: Causes growth arrest of the procyclic form.
FT Complete loss of catalytic activity; when associated with
FT 2-N--H-25 DEL."
FT /evidence="ECO:0000269|PubMed:19465686,
FT ECO:0000269|PubMed:20403364"
FT MUTAGEN 141
FT /note="N->A: Partial loss of catalytic activity; when
FT associated with 2-N--H-25 DEL."
FT /evidence="ECO:0000269|PubMed:20403364"
FT MUTAGEN 181
FT /note="N->A,D: Complete loss of catalytic activity; when
FT associated with 2-N--H-25 DEL."
FT /evidence="ECO:0000269|PubMed:20403364"
FT MUTAGEN 183
FT /note="F->A: Complete loss of catalytic activity; when
FT associated with 2-N--H-25 DEL."
FT /evidence="ECO:0000269|PubMed:20403364"
FT MUTAGEN 335
FT /note="D->A: Complete loss of catalytic activity; when
FT associated with 2-N--H-25 DEL."
FT /evidence="ECO:0000269|PubMed:20403364"
FT MUTAGEN 338
FT /note="N->A: Partial loss of catalytic activity; when
FT associated with 2-N--H-25 DEL."
FT /evidence="ECO:0000269|PubMed:20403364"
FT MUTAGEN 342
FT /note="C->A: No effect on catalytic activity; when
FT associated with 2-N--H-25 DEL."
FT /evidence="ECO:0000269|PubMed:20403364"
FT MUTAGEN 344
FT /note="R->A: Partial loss of catalytic activity; when
FT associated with 2-N--H-25 DEL."
FT /evidence="ECO:0000269|PubMed:20403364"
FT MUTAGEN 345
FT /note="R->A: Complete loss of catalytic activity; when
FT associated with 2-N--H-25 DEL."
FT /evidence="ECO:0000269|PubMed:20403364"
SQ SEQUENCE 406 AA; 46764 MW; 5840BE19ED366A1E CRC64;
MNVAKREFIR GMMAHYRASL PPPEHSVVIH ELQKRVLDIG MLAVNKAHVE LFGSHVSGFC
TPHSDADISL TYRNFSPWLQ GMERVDEQNN KRMTRFGKEA SAMGMEDVRY IRARIPVVQF
TDGVTGIHCD VSIGNIGGVE NSKILCAIRQ VFPDFYGAYI HLVKAWGKAR EVIAPERSTF
NSFTVTTMAL MVLQELGLLP VFSKPTGEFG ELTVADAEML LQEFKLPPIY DSLHDDDEKL
GEAVFFCLQR FAEYYAKYDF SAGTVSLIHP RRHRTVYERV VRRHLELLGS RKRLEWEKHI
AEHKEDGPLD ENDFSASMQN ETTQRPSNSP YVVEDFVNYV NCGRRVQASR VRHIQQEFNR
LREMLIDKES ELKFDEVFRE SDTVPRFQGF EGVGTRDHRV KTFRPQ
