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Information on EC 2.7.7.19 - polynucleotide adenylyltransferase and Organism(s) Escherichia coli and UniProt Accession P0ABF1
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2.7.7.19 polynucleotide adenylyltransferaseIUBMB Comments
Also acts slowly with CTP. Catalyses template-independent extension of the 3'- end of a DNA strand by one nucleotide at a time. Cannot initiate a chain de novo. The primer, depending on the source of the enzyme, may be an RNA or DNA fragment, or oligo(A) bearing a 3'-OH terminal group. See also EC 2.7.7.6 DNA-directed RNA polymerase.
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Word Map
- 2.7.7.19
-
polyadenylation
- polymerases
- pre-mrnas
- phosphorylase
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exosome
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rna-binding
- vaccinia
- trna
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oligoa
- helicase
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tramp
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aauaaa
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deadenylation
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polya-binding
- pnpase
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dna-dependent
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meiotic
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noncanonical
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exonuclease
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3'-terminal
- cordycepin
-
oligoadenylation
- exoribonucleases
- polyu
- drug development
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endoribonuclease
- medicine
- snornas
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non-templated
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cca-adding
- hfq
-
polya-specific
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deadenylase
- biotechnology
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polya-tailed
-
snrnp
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pabpn1
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degradosome
-
exonucleolytic
- 3'-deoxyadenosine
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pancreatitis-associated
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
poly(a) polymerase, pap i, fam46c, gld-2, star-pap, fam46a, poly(a) polymerase i, pap ii, papd5, mtpap, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
adenosine triphosphate:ribonucleic acid adenylyltransferase
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-
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AMP polynucleotidylexotransferase
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-
-
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ATP-polynucleotide adenylyltransferase
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-
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ATP:polynucleotidylexotransferase
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-
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neo-PAP
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-
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NTP polymerase
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-
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nucleotidyltransferase, polyadenylate
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-
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PAP
PAP I
poly(A) hydrolase
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-
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poly(A) polymerase
poly(A) synthetase
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-
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polyadenylate nucleotidyltransferase
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-
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polyadenylate polymerase
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-
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polyadenylate synthetase
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-
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polyadenylic acid polymerase
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-
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polyadenylic polymerase
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-
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RNA adenylating enzyme
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-
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RNA formation factors, PF1
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-
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terminal riboadenylate transferase
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-
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PAP
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-
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PAP I
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-
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poly(A) polymerase
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-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
nucleotidyl group transfer
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SYSTEMATIC NAME
IUBMB Comments
ATP:polynucleotide adenylyltransferase
Also acts slowly with CTP. Catalyses template-independent extension of the 3'- end of a DNA strand by one nucleotide at a time. Cannot initiate a chain de novo. The primer, depending on the source of the enzyme, may be an RNA or DNA fragment, or oligo(A) bearing a 3'-OH terminal group. See also EC 2.7.7.6 DNA-directed RNA polymerase.
CAS REGISTRY NUMBER
COMMENTARY
9026-30-6
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + adenosine(5')pentaphospho(5')adenosine
diphosphate + ?
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i.e. AP5A
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-
?
ATP + adenosine(5')tetraphospho(5')adenosine
diphosphate + ?
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i.e. AP4A
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-
?
ATP + adenosine(5')triphospho(5')adenosine
diphosphate + ?
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i.e. AP3A
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-
?
ATP + guanosine(5')pentaphospho(5')guanosine
diphosphate + ?
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i.e. GP5G
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-
?
ATP + guanosine(5')tetraphospho(5')guanosine
diphosphate + ?
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i.e. GP4G
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-
?
ATP + guanosine(5')triphospho(5')guanosine
diphosphate + ?
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i.e. GP3G
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-
?
ATP + RNA
diphosphate + RNA(A)n
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primers: poly(U), poly(C), poly(A), not poly(G)
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-
?
ATP + RNA
diphosphate + RNA(A)n
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Mn2+-activated enzymes are indifferent to primer length
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?
ATP + RNA
diphosphate + RNA(A)n
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no specificity for the 3'-terminal nucleotides
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?
ATP + RNA
diphosphate + RNA(A)n
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enzyme uses all four nucleoside triphosphates
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-
?
ATP + RNA
diphosphate + RNA(A)n
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other nucleotides polymerized at less than 1% of the ATP rate
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?
ATP + RNA
diphosphate + RNA(A)n
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enzyme is unable to catalyze pyrophosphorolysis or phosphorolysis reaction
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?
ATP + RNA
diphosphate + RNA(A)n
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rather low specificity for primer
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?
ATP + RNAn
diphosphate + RNAn+1
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preferentially elongates RNA harbouring poly(A) tails bound by Hfq
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?
additional information
?
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rRNA fragments and tRNA precursors originating from the internal spacer regions of the rrn operons, in particular, rrnB are abundant poly(A) polymerase targets. Glu tRNA precursors originating from the rrnB and rrnG transcripts exhibit long 3' trailers that are primarily removed by polyribonucleotide nucleotidyltransferase and to a lesser extent by RNase II and poly(A) polymerase. Glu tRNA precursors still harbouring the 5' leader can be degraded by a 3' to 5' quality control pathway involving poly(A) polymerase
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?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
rRNA fragments and tRNA precursors originating from the internal spacer regions of the rrn operons, in particular, rrnB are abundant poly(A) polymerase targets. Glu tRNA precursors originating from the rrnB and rrnG transcripts exhibit long 3' trailers that are primarily removed by polyribonucleotide nucleotidyltransferase and to a lesser extent by RNase II and poly(A) polymerase. Glu tRNA precursors still harbouring the 5' leader can be degraded by a 3' to 5' quality control pathway involving poly(A) polymerase
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?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Mn2+
additional information
-
overview: ion requirements, poly(A) polymerases purified from different sources, and in some cases even from the same source, respond differently to the presence of Mg2+ and Mn2+
Mg2+
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divalent cation requirement may be fulfilled by Mn2+, Mg2+ or a combination of the two depending on the source of the enzyme
Mg2+
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in presence of Mg2+ and a specificity factor required for correct cleavage at the poly(A) site of pre-mRNA
Mn2+
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divalent cation requirement may be fulfilled by Mn2+, Mg2+ or a combination of the two depending on the source of the enzyme
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
stem-loop structure in mRNA 3-ends may be inhibitory
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
host factor I
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Hfq-PAP I interaction facilitates RNA recognition by PAP I
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
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PAP I leader sequence directs the protein to the cellular membrane
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
mature tRNAs, which are normally not substrates for PAP I in wild-type cells, are rapidly polyadenylated as PAP I levels increase upon overexpression, leading to dramatic reductions in the fraction of aminoacylated tRNAs, cessation of protein synthesis and cell death. The toxicity associated with PAP I is exacerbated by the absence of either RNase T and/or RNase PH. Regulation of PAP I is critical not for preventing the decay of mRNAs, but rather for maintaining normal levels of functional tRNAs and protein synthesis
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50000
58000
additional information
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heterogenous, monomers to very large aggregates, all forms being active, gel filtration, recombinant protein
50000
-
x * 50000, heterogenous, monomers to very large aggregates, all forms being active, SDS-PAGE, gel filtration, recombinant protein
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
monomer
?
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x * 50000, heterogenous, monomers to very large aggregates, all forms being active, SDS-PAGE, gel filtration, recombinant protein
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
phosphoprotein
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enzyme PAP I variant with C-terminal His-tag can be phosphorylated both in vivo and in vitro. In vivo phosphorylation impairs activity of the enzyme and may be a regulatory process
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D212A
mutant enzyme forms specifically incorporated A-residues with a fidelity comparable to that of the wild type enzyme
E211A
mutant enzyme forms specifically incorporated A-residues with a fidelity comparable to that of the wild type enzyme
R215A
the mutation leads to a dramatic loss in nucleotide specificity and the formation of poly(N) tails
D170A
-
diphosphate release equivalent to AMP production, 5-30% of wild type activity
D170P
D214A
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diphosphate release equivalent to AMP production, 5-30% of wild type activity
D214P
D88A
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diphosphate release equivalent to AMP production, 5-30% of wild type activity
D88P
D90A
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diphosphate release equivalent to AMP production, 5-30% of wild type activity
D90P
additional information
D170P
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diphosphate release equivalent to AMP production, 5-30% of wild type activity
D214P
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diphosphate release equivalent to AMP production, 5-30% of wild type activity
D88P
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diphosphate release equivalent to AMP production, 5-30% of wild type activity
D90P
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diphosphate release equivalent to AMP production, 5-30% of wild type activity
additional information
-
chimeras of CCA-adding enzyme and PAP were constructed
additional information
-
enzyme knockout mutant, the mRNA levels of bolA, which is induced in response to many forms of stress, are reduced 2.5fold in stationary phase. Absence of enzyme enhances the RssB-mediated deltaS proteolysis specifically in starved cells
additional information
-
enzyme PAP I variant with C-terminal His-tag can be phosphorylated both in vivo and in vitro. In vivo phosphorylation impairs activity of the enzyme
additional information
-
reengineering of enzyme in order to function as (UG) adding enzyme. Double- and triple-mutants in residues 211, 212, 215 add 570 G residues to oligoA15 substrate
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
The wild-type poly(A) polymerase, as well as deletion variants are expressed in Escherichia coli BL21(DE3) (Novagen). Freshly transformed cells are grown at 28-37°C in 700 ml LB medium containing 0.030 mg/ml kanamycin and 0.033 mg/ml chloramphenicol. At mid-log phase (A600 = 0.6), expression is induced by the addition of IPTG to a final concentration of 0.200 mM. After 3-5 hr of incubation at 28-37°C, cells are harvested by centrifugation and lysed by lysozyme treatment and sonication in ice-cold buffer I (20 mM Tris/HCl pH 7.6, 0.5 M NaCl, 5 mM imidazole and 0.75 mg/ml lysozyme). After centrifugation for 30 min at 25000 xg and 4°C, the proteins in the supernatant are purified by FPLC on a 5 ml HiTrap Chelating Sepharose column (Amersham Biosciences) and eluted with 500 mM imidazole. Fractions containing the enzymes as determined by SDS-PAGE are pooled, dialyzed against buffer II (20 mM Tris/HCl pH 7.6, 0.5 M NaCl, 5 mM MgCl2 and 10% glycerol). All proteins are stored in the presence of 40% (v/v) glycerol at -20°C.
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Edmonds, M.
Poly(A) adding enzymes
The Enzymes,3rd Ed. (Boyer,P. D. ,ed. )
15
217-244
1982
Bos taurus, Cricetulus griseus, Coturnix sp., Escherichia coli, Homo sapiens, Mus musculus, Rattus norvegicus, Vaccinia virus
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Sippel, A.E.
Purification and characterization of adenosine triphosphate: ribonucleic acid adenyltransferase from Escherichia coli
Eur. J. Biochem.
37
31-40
1973
Escherichia coli
Raynal, L.C.; Krisch, H.M.; Carpousis, A.J.
Bacterial poly(A) polymerase: an enzyme that modulates RNA stability
Biochimie
78
390-398
1996
Escherichia coli
Raynal, L.C.; Carpousis, A.J.
Poly(A) polymerase I of Escherichia coli: characterization of the catalytic domain, an RNA binding site and regions for the interaction with proteins involved in mRNA degradation
Mol. Microbiol.
32
765-775
1999
Escherichia coli
Yehudai-Resheff, S.; Schuster, G.
Characterization of the E.coli poly(A) polymerase: nucleotide specificity, RNA-binding affinities and RNA structure dependence
Nucleic Acids Res.
28
1139-1144
2000
Escherichia coli
Sillero, M.A.; Socorro, S.; Baptista, M.J.; Del Valle, M.; De Diego, A.; Sillero, A.
Poly(A) polymerase from Escherichia coli adenylylates the 3'-hydroxyl residue of nucleosides, nucleoside 5'-phosphates and nucleoside(5')oligophospho(5')nucleosides (NpnN)
Eur. J. Biochem.
268
3605-3611
2001
Escherichia coli
Betat, H.; Rammelt, C.; Martin, G.; Moerl, M.
Exchange of regions between bacterial poly(A) polymerase and the CCA-adding enzyme generates altered specificities
Mol. Cell
15
389-398
2004
Escherichia coli
Jasiecki, J.; Wegrzyn, G.
Localization of Escherichia coli poly(A) polymerase I in cellular membrane
Biochem. Biophys. Res. Commun.
329
598-602
2005
Escherichia coli
Folichon, M.; Allemand, F.; Regnier, P.; Hajnsdorf, E.
Stimulation of poly(A) synthesis by Escherichia coli poly(A)polymerase I is correlated with Hfq binding to poly(A) tails
FEBS J.
272
454-463
2005
Escherichia coli
Jasiecki, J.; Wegrzyn, G.
Phosphorylation of Escherichia coli poly(A) polymerase I and effects of this modification on the enzyme activity
FEMS Microbiol. Lett.
261
118-122
2006
Escherichia coli
Santos, J.M.; Freire, P.; Mesquita, F.S.; Mika, F.; Hengge, R.; Arraiano, C.M.
Poly(A)-polymerase I links transcription with mRNA degradation via sigmaS proteolysis
Mol. Microbiol.
60
177-188
2006
Escherichia coli
Cho, H.D.; Verlinde, C.L.; Weiner, A.M.
Reengineering CCA-adding enzymes to function as (U,G)- or dCdCdA-adding enzymes or poly(C,A) and poly(U,G) polymerases
Proc. Natl. Acad. Sci. USA
104
54-59
2007
Escherichia coli
Just, A.; Butter, F.; Trenkmann, M.; Heitkam, T.; Moerl, M.; Betat, H.
A comparative analysis of two conserved motifs in bacterial poly(A) polymerase and CCA-adding enzyme
Nucleic Acids Res.
36
5212-5220
2008
Escherichia coli (P0ABF1), Escherichia coli
Maes, A.; Gracia, C.; Hajnsdorf, E.; Regnier, P.
Search for poly(A) polymerase targets in E. coli reveals its implication in surveillance of Glu tRNA processing and degradation of stable RNAs
Mol. Microbiol.
83
436-451
2012
Escherichia coli
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