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[1]
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NUCLEOTIDE SEQUENCE [GENOMIC DNA].
PubMed=3009457 [NCBI, ExPASy, EBI, Israel, Japan]
Cudny H.,
Lupski J.R.,
Godson G.N.,
Deutscher M.P.;
"Cloning, sequencing, and species relatedness of the Escherichia coli cca gene encoding the enzyme tRNA nucleotidyltransferase.";
J. Biol. Chem. 261:6444-6449(1986).
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[2]
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NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
STRAIN=K12 / MG1655 / ATCC 47076;
DOI=10.1126/science.277.5331.1453; PubMed=9278503 [NCBI, ExPASy, EBI, Israel, Japan]
Blattner F.R.,
Plunkett G. III,
Bloch C.A.,
Perna N.T.,
Burland V.,
Riley M.,
Collado-Vides J.,
Glasner J.D.,
Rode C.K.,
Mayhew G.F.,
Gregor J.,
Davis N.W.,
Kirkpatrick H.A.,
Goeden M.A.,
Rose D.J.,
Mau B.,
Shao Y.;
"The complete genome sequence of Escherichia coli K-12.";
Science 277:1453-1474(1997).
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[3]
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NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
DOI=10.1038/msb4100049; PubMed=16738553 [NCBI, ExPASy, EBI, Israel, Japan]
Hayashi K.,
Morooka N.,
Yamamoto Y.,
Fujita K.,
Isono K.,
Choi S.,
Ohtsubo E.,
Baba T.,
Wanner B.L.,
Mori H.,
Horiuchi T.;
"Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110.";
Mol. Syst. Biol. 2:E1-E5(2006).
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[4]
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NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 65-76, AND MUTAGENESIS OF GLY-70.
PubMed=3533927 [NCBI, ExPASy, EBI, Israel, Japan]
Zhu L.Q.,
Cudny H.,
Deutscher M.P.;
"A mutation in Escherichia coli tRNA nucleotidyltransferase that affects only AMP incorporation is in a sequence often associated with nucleotide-binding proteins.";
J. Biol. Chem. 261:14875-14877(1986).
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[5]
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FUNCTION, NUCLEOTIDYLTRANSFERASE ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
PubMed=3516995 [NCBI, ExPASy, EBI, Israel, Japan]
Cudny H.,
Deutscher M.P.;
"High-level overexpression, rapid purification, and properties of Escherichia coli tRNA nucleotidyltransferase.";
J. Biol. Chem. 261:6450-6453(1986).
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[6]
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PHOSPHOHYDROLASE ACTIVITIES, CHARACTERIZATION, SUBUNIT, BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF ASP-21; ASP-23; HIS-255; ASP-256; HIS-305 AND ASP-306.
DOI=10.1074/jbc.M405120200; PubMed=15210699 [NCBI, ExPASy, EBI, Israel, Japan]
Yakunin A.F.,
Proudfoot M.,
Kuznetsova E.,
Savchenko A.,
Brown G.,
Arrowsmith C.H.,
Edwards A.M.;
"The HD domain of the Escherichia coli tRNA nucleotidyltransferase has 2',3'-cyclic phosphodiesterase, 2'-nucleotidase, and phosphatase activities.";
J. Biol. Chem. 279:36819-36827(2004).
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- FUNCTION: Catalyzes the addition and repair of the essential 3'-terminal CCA sequence in tRNAs without using a nucleic acid template. Adds these three nucleotides in the order of C, C, and A to the tRNA nucleotide-73, using CTP and ATP as substrates and producing inorganic pyrophosphate. Also shows highest phosphatase activity in the presence of Ni(2+) and hydrolyzes pyrophosphate, canonical 5'-nucleoside tri- and diphosphates, NADP, and 2'-AMP with the production of Pi. Displays a metal-independent phosphodiesterase activity toward 2',3'-cAMP, 2',3'-cGMP, and 2',3'-cCMP. Without metal or in the presence of Mg(2+), this protein hydrolyzes 2',3'-cyclic substrates with the formation of 2'-nucleotides, whereas in the presence of Ni(2+), it also produces some 3'-nucleotides. These phosphohydrolase activities are probably involved in the repair of the tRNA 3'-CCA terminus degraded by intracellular RNases.
- CATALYTIC ACTIVITY: ATP + tRNA(n) = diphosphate + tRNA(n+1).
- CATALYTIC ACTIVITY: CTP + tRNA(n) = diphosphate + tRNA(n+1).
- COFACTOR: Magnesium for nucleotidyltransferase activity.
- COFACTOR: Nickel for phosphatase activity.
- ENZYME REGULATION: Both phosphatase and phosphodiesterase activities are competitively inhibited by low concentrations of the E.coli tRNA (10 nM). Cu(2+) stimulates the hydrolysis of pyrophosphate and ATP and completely inhibits the hydrolysis of 2'-AMP. The phosphodiesterase activity is inhibited by Zn(2+), Cu(2+) and Co(2+).
- BIOPHYSICOCHEMICAL PROPERTIES:
| Kinetic parameters: |
KM=0.33 mM for ATP (in the tRNA-NT activity assay); | | KM=0.03 mM for CTP (in the tRNA-NT activity assay); | | KM=0.015 mM for tRNA-CC; | | KM=0.02 mM for tRNA-C; | | KM=6.2 mM for pNPP; | | KM=0.10 mM for PPi; | | KM=0.15 mM for NADP; | | KM=0.19 mM for ADP; | | KM=0.18 mM for ATP (in the phosphatase activity assay); | | KM=0.53 mM for CDP; | | KM=0.13 mM for CTP (in the phosphatase activity assay); | | KM=0.76 mM for 2'-AMP; | | KM=0.49 mM for 2',3'-cAMP; | | KM=1.60 mM for 2',3'-cGMP; | | Vmax=12.4 µmol/min/mg enzyme with pNPP as substrate; | | Vmax=3.01 µmol/min/mg enzyme with PPi as substrate; | | Vmax=17.9 µmol/min/mg enzyme with NADP as substrate; | | Vmax=1.49 µmol/min/mg enzyme with ADP as substrate; | | Vmax=4.53 µmol/min/mg enzyme with ATP as substrate (in the phosphatase activity assay); | | Vmax=5.80 µmol/min/mg enzyme with CDP as substrate; | | Vmax=4.03 µmol/min/mg enzyme with CTP as substrate (in the phosphatase activity assay); | | Vmax=3.71 µmol/min/mg enzyme with 2'-AMP as substrate; | | Vmax=3.21 µmol/min/mg enzyme with 2',3'-cAMP as substrate; | | Vmax=2.36 µmol/min/mg enzyme with 2',3'-cGMP as substrate; | | pH dependence: |
Optimum pH is 9.4 for AMP incorporation, 10.0 for CMP incorporation, and 7.0 for the phosphatase and phosphodiesterase activities; | |
- SUBUNIT: Monomer. Can also form homodimers and oligomers, but with low levels.
- INTERACTION:
P0A6M4:dtd; NbExp=1; IntAct=EBI-545256, EBI-562575;
- DOMAIN: Comprises two domains: an N-terminal domain containing the nucleotidyltransferase activity and a C-terminal HD domain associated with both phosphodiesterase and phosphatase activities.
- MISCELLANEOUS: A single active site specifically recognizes both ATP and CTP and is responsible for their addition (By similarity).
- SIMILARITY: Belongs to the tRNA nucleotidyltransferase/poly(A) polymerase family. Bacterial CCA-adding enzyme type 1 subfamily.
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Distributed under the Creative Commons Attribution-NoDerivs License.
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| Length: 412 AA [This is the length of the unprocessed precursor] |
Molecular weight: 46467 Da [This is the MW of the unprocessed precursor] |
CRC64: 947182E6086220F7 [This is a checksum on the sequence] |
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10 20 30 40 50 60
MKIYLVGGAV RDALLGLPVK DRDWVVVGST PQEMLDAGYQ QVGRDFPVFL HPQTHEEYAL
70 80 90 100 110 120
ARTERKSGSG YTGFTCYAAP DVTLEDDLKR RDLTINALAQ DDNGEIIDPY NGLGDLQNRL
130 140 150 160 170 180
LRHVSPAFGE DPLRVLRVAR FAARYAHLGF RIADETLALM REMTHAGELE HLTPERVWKE
190 200 210 220 230 240
TESALTTRNP QVFFQVLRDC GALRVLFPEI DALFGVPAPA KWHPEIDTGI HTLMTLSMAA
250 260 270 280 290 300
MLSPQVDVRF ATLCHDLGKG LTPPELWPRH HGHGPAGVKL VEQLCQRLRV PNEIRDLARL
310 320 330 340 350 360
VAEFHDLIHT FPMLNPKTIV KLFDSIDAWR KPQRVEQLAL TSEADVRGRT GFESADYPQG
370 380 390 400 410
RWLREAWEVA QSVPTKAVVE AGFKGVEIRE ELTRRRIAAV ASWKEQRCPK PE
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P06961 in FASTA format |
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