CCA_SACSH
ID CCA_SACSH Reviewed; 412 AA.
AC P77978;
DT 15-DEC-1998, integrated into UniProtKB/Swiss-Prot.
DT 16-NOV-2001, sequence version 2.
DT 03-AUG-2022, entry version 89.
DE RecName: Full=CCA-adding enzyme;
DE EC=2.7.7.72;
DE AltName: Full=CCA tRNA nucleotidyltransferase;
DE AltName: Full=tRNA CCA-pyrophosphorylase;
DE AltName: Full=tRNA adenylyl-/cytidylyl- transferase;
DE AltName: Full=tRNA nucleotidyltransferase;
DE AltName: Full=tRNA-NT;
GN Name=cca;
OS Saccharolobus shibatae (Sulfolobus shibatae).
OC Archaea; Crenarchaeota; Thermoprotei; Sulfolobales; Sulfolobaceae;
OC Saccharolobus.
OX NCBI_TaxID=2286;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND BIOPHYSICOCHEMICAL
RP PROPERTIES.
RX PubMed=8809016;
RA Yue D., Maizels N., Weiner A.M.;
RT "CCA-adding enzymes and poly(A) polymerases are all members of the same
RT nucleotidyltransferase superfamily: characterization of the CCA-adding
RT enzyme from the archaeal hyperthermophile Sulfolobus shibatae.";
RL RNA 2:895-908(1996).
RN [2]
RP SEQUENCE REVISION TO 349-363.
RA Cho H.D., Weiner A.M.;
RL Submitted (NOV-2001) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP MUTAGENESIS OF ASP-53; ASP-55; ASP-106; GLU-173 AND ASP-215.
RX PubMed=9792681; DOI=10.1074/jbc.273.45.29693;
RA Yue D., Weiner A.M., Maizels N.;
RT "The CCA-adding enzyme has a single active site.";
RL J. Biol. Chem. 273:29693-29700(1998).
RN [4]
RP SUBUNIT.
RX PubMed=11090289; DOI=10.1006/jmbi.2000.4189;
RA Li F., Wang J., Steitz T.A.;
RT "Sulfolobus shibatae CCA-adding enzyme forms a tetramer upon binding two
RT tRNA molecules: a scrunching-shuttling model of CCA specificity.";
RL J. Mol. Biol. 304:483-492(2000).
RN [5]
RP MECHANISM.
RX PubMed=15265870; DOI=10.1074/jbc.m405518200;
RA Cho H.D., Weiner A.M.;
RT "A single catalytically active subunit in the multimeric Sulfolobus
RT shibatae CCA-adding enzyme can carry out all three steps of CCA addition.";
RL J. Biol. Chem. 279:40130-40136(2004).
RN [6]
RP 3D-STRUCTURE MODELING, AND MUTAGENESIS OF HIS-93; TYR-95; HIS-129 AND
RP ARG-221.
RX PubMed=15590678; DOI=10.1074/jbc.m412594200;
RA Cho H.D., Verlinde C.L., Weiner A.M.;
RT "Archaeal CCA-adding enzymes: central role of a highly conserved beta-turn
RT motif in RNA polymerization without translocation.";
RL J. Biol. Chem. 280:9555-9566(2005).
CC -!- FUNCTION: Catalyzes the addition and repair of the essential 3'-
CC terminal CCA sequence in tRNAs without using a nucleic acid template.
CC Adds these three nucleotides in the order of C, C, and A to the tRNA
CC nucleotide-73, using CTP and ATP as substrates and producing inorganic
CC pyrophosphate. {ECO:0000269|PubMed:8809016}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a tRNA precursor + ATP + 2 CTP = a tRNA with a 3' CCA end + 3
CC diphosphate; Xref=Rhea:RHEA:14433, Rhea:RHEA-COMP:10465, Rhea:RHEA-
CC COMP:10468, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:37563,
CC ChEBI:CHEBI:74896, ChEBI:CHEBI:83071; EC=2.7.7.72;
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; Evidence={ECO:0000250};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=30 uM for CTP {ECO:0000269|PubMed:8809016};
CC Note=The KM for ATP is much higher than 30 uM.;
CC pH dependence:
CC Optimum pH is about 9. Active from pH 7 to 10.
CC {ECO:0000269|PubMed:8809016};
CC Temperature dependence:
CC Optimum temperature is 70 degrees Celsius. Inactive at 37 degrees
CC Celsius. {ECO:0000269|PubMed:8809016};
CC -!- SUBUNIT: Homodimer. Forms a tetramer upon binding two tRNAs. However,
CC tRNA-induced tetramer formation is not required for CCA addition.
CC {ECO:0000269|PubMed:11090289}.
CC -!- MISCELLANEOUS: A single active site specifically recognizes both ATP
CC and CTP and is responsible for their addition.
CC -!- SIMILARITY: Belongs to the tRNA nucleotidyltransferase/poly(A)
CC polymerase family. Archaeal CCA-adding enzyme subfamily. {ECO:0000305}.
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DR EMBL; U66004; AAB53697.2; -; Genomic_DNA.
DR PIR; A59397; T48856.
DR AlphaFoldDB; P77978; -.
DR SMR; P77978; -.
DR BRENDA; 2.7.7.72; 6162.
DR SABIO-RK; P77978; -.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-UniRule.
DR GO; GO:0052929; F:ATP:3'-cytidine-cytidine-tRNA adenylyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0052928; F:CTP:3'-cytidine-tRNA cytidylyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0052927; F:CTP:tRNA cytidylyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0000287; F:magnesium ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0000049; F:tRNA binding; IEA:UniProtKB-UniRule.
DR GO; GO:0016437; F:tRNA cytidylyltransferase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0042245; P:RNA repair; IEA:UniProtKB-KW.
DR GO; GO:0001680; P:tRNA 3'-terminal CCA addition; IEA:UniProtKB-UniRule.
DR CDD; cd05400; NT_2-5OAS_ClassI-CCAase; 1.
DR Gene3D; 1.10.1410.30; -; 1.
DR Gene3D; 3.30.460.10; -; 1.
DR HAMAP; MF_01264; CCA_arch; 1.
DR InterPro; IPR008229; CCA-adding_arc.
DR InterPro; IPR042090; CCA_tRNA_nucleotrans_2.
DR InterPro; IPR006116; NT_2-5OAS_ClassI-CCAase.
DR InterPro; IPR043519; NT_sf.
DR InterPro; IPR011068; NuclTrfase_I-like_C.
DR InterPro; IPR002934; Polymerase_NTP_transf_dom.
DR InterPro; IPR015329; tRNA_NucTransf2.
DR PANTHER; PTHR39643; PTHR39643; 1.
DR Pfam; PF01909; NTP_transf_2; 1.
DR Pfam; PF09249; tRNA_NucTransf2; 1.
DR PIRSF; PIRSF005335; CCA_arch; 1.
DR SUPFAM; SSF55003; SSF55003; 1.
DR SUPFAM; SSF81301; SSF81301; 1.
DR TIGRFAMs; TIGR03671; cca_archaeal; 1.
PE 1: Evidence at protein level;
KW ATP-binding; Magnesium; Metal-binding; Nucleotide-binding;
KW Nucleotidyltransferase; RNA repair; RNA-binding; Transferase;
KW tRNA processing.
FT CHAIN 1..412
FT /note="CCA-adding enzyme"
FT /id="PRO_0000139082"
FT BINDING 41
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01264"
FT BINDING 41
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01264"
FT BINDING 44
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01264"
FT BINDING 44
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01264"
FT BINDING 53
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250"
FT BINDING 55
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250"
FT BINDING 106
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250"
FT BINDING 129
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01264"
FT BINDING 129
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01264"
FT BINDING 149
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01264"
FT BINDING 149
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01264"
FT BINDING 158
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01264"
FT BINDING 158
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01264"
FT MUTAGEN 53
FT /note="D->A: Loss of both AMP and CMP incorporation."
FT /evidence="ECO:0000269|PubMed:9792681"
FT MUTAGEN 55
FT /note="D->A,E: Loss of both AMP and CMP incorporation."
FT /evidence="ECO:0000269|PubMed:9792681"
FT MUTAGEN 93
FT /note="H->V: Loss of AMP incorporation but no loss of CMP
FT incorporation."
FT /evidence="ECO:0000269|PubMed:15590678"
FT MUTAGEN 95
FT /note="Y->V: Adds C75 and A76 but not C74."
FT /evidence="ECO:0000269|PubMed:15590678"
FT MUTAGEN 106
FT /note="D->A: High decrease in AMP incorporation but not in
FT CMP incorporation."
FT /evidence="ECO:0000269|PubMed:9792681"
FT MUTAGEN 129
FT /note="H->V: Loss of CMP incorporation but no loss of AMP
FT incorporation."
FT /evidence="ECO:0000269|PubMed:15590678"
FT MUTAGEN 173
FT /note="E->A: High decrease in both AMP and CMP
FT incorporation."
FT /evidence="ECO:0000269|PubMed:9792681"
FT MUTAGEN 215
FT /note="D->A: High decrease in both AMP and CMP
FT incorporation."
FT /evidence="ECO:0000269|PubMed:9792681"
FT MUTAGEN 221
FT /note="R->A,E: High decrease in both AMP and CMP
FT incorporation."
FT /evidence="ECO:0000269|PubMed:15590678"
SQ SEQUENCE 412 AA; 47848 MW; 745E1E9A8816B43C CRC64;
MIEEEVLKII KPTEEDKKGI EKVLEIIRER LNKLDFEVEG SFRKGTWLRQ DTDVDVFVFY
PKDVGKEYLE RNALNDIINR IKDLDYTLAY AEHPYVIVNI NNVEVDIVPA LRVESGDRAI
TAVDRTPFHT KYVTSHLDER GKDEVRLLKR FMKGIGVYGA ELKVQGFSGY ATELLVIYYG
SFRKVLEAAS KWKHPIKIEL TKPMRAFSEP LIIPDPVDPR RNVTAAVSLK NIATFSVAAK
YYLKNPSMEF FFPSKKVEEK IKGDVLILRL NLDEKSSEDI IWGQIKRSVN KIERALKQSG
FRVIDIQAWG DTSNIVIAVQ LESKNIGQYY LNIGPQYYSE TIDDFIQKND NIWVGEDGRL
YSIKERKEYN AEAIAKKNIV LKVKYNIESY WLQNTEDQQI MKFLRKTPTW LK
