Information on EC 6.5.1.3 - RNA ligase (ATP)

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
6.5.1.3
-
RECOMMENDED NAME
GeneOntology No.
RNA ligase (ATP)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(ribonucleotide)n-3'-hydroxyl + 5'-(5'-diphosphoadenosine)-(ribonucleotide)m = (ribonucleotide)n+m + AMP
show the reaction diagram
(1c)
-
-
-
ATP + (ribonucleotide)n-3'-hydroxyl + 5'-phospho-(ribonucleotide)m = (ribonucleotide)n+m + AMP + diphosphate
show the reaction diagram
ATP + [RNA ligase]-L-lysine = [RNA ligase]-N6-(5'-adenylyl)-L-lysine + diphosphate
show the reaction diagram
(1a)
-
-
-
[RNA ligase]-N6-(5'-adenylyl)-L-lysine + 5'-phospho-(ribonucleotide)m = 5'-(5'-diphosphoadenosine)-(ribonucleotide)m + [RNA ligase]-L-lysine
show the reaction diagram
(1b)
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
esterification
-
-
phosphodiester
-
SYSTEMATIC NAME
IUBMB Comments
poly(ribonucleotide)-3'-hydroxyl:5'-phospho-poly(ribonucleotide) ligase (ATP)
The enzyme catalyses the ligation of RNA strands with 3'-hydroxyl and 5'-phosphate termini, forming a phosphodiester and sealing certain types of single-strand breaks in RNA. Catalysis occurs by a three-step mechanism, starting with the activation of the enzyme by ATP, forming a phosphoramide bond between adenylate and a lysine residue. The adenylate group is then transferred to the 5'-phosphate terminus of the substrate, forming the capped structure 5'-(5'-diphosphoadenosine)-[RNA]. Finally, the enzyme catalyses a nucleophilic attack of the 3'-OH terminus on the capped terminus, which results in formation of the phosphodiester bond and release of the adenylate.
CAS REGISTRY NUMBER
COMMENTARY hide
37353-39-2
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain PCC 7120
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
thermophilic bacteriophage that infects the thermophilic eubacterium Rhodothermus marinus
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-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(deoxyribonucleotide)n
circular (deoxyribonucleotide)n
show the reaction diagram
-
-
-
-
?
(ribonucleotide)n
circular (ribonucleotide)n
show the reaction diagram
-
-
-
-
?
5'-adenylated cytidine 5',3'-bisphosphate + diguanosine tetraphosphate
Gp4GpCp + pCpGp4GpCp
show the reaction diagram
-
-
-
?
8-Bromoadenosine 5'-triphosphate + (ribonucleotide)n + (ribonucleotide)m
8-Bromoadenosine monophosphate + (ribonucleotide)n+m
show the reaction diagram
-
-
-
-
-
8-Sulfoadenosine 5'-triphosphate + (ribonucleotide)n + (ribonucleotide)m
8-Sulfoadenosinemonophosphate + phosphate + (ribonucleotide)n+m
show the reaction diagram
-
-
-
-
-
ATP + (2E,6E)-farnesyl diphosphate
(2E,6E)-farnesyl-triphospho(5')adenosine + diphosphate
show the reaction diagram
-
assay contains pyruvate kinase, phosphoenolpyruvate and pyrophosphatase to recover ATP
-
-
ir
ATP + (2E,6E)-farnesyl triphosphate
(2E,6E)-farnesyl-tetraphospho(5')adenosine + diphosphate
show the reaction diagram
-
assay contains pyruvate kinase, phosphoenolpyruvate and pyrophosphatase to recover ATP
-
-
ir
ATP + (deoxyribonucleotide)n
adenylyl-(deoxyribonucleotide)n + diphosphate
show the reaction diagram
-
-
-
-
?
ATP + (ribonucleotide)n
adenylyl-(ribonucleotide)n + diphosphate
show the reaction diagram
-
-
-
-
?
ATP + (ribonucleotide)n + (ribonucleotide)m
?
show the reaction diagram
ATP + (ribonucleotide)n + (ribonucleotide)m
AMP + diphosphate + (ribonucleotide)n+m
show the reaction diagram
ATP + 5'-aaaCCAGUC
AMP + diphosphate + ?
show the reaction diagram
-
RNA-DNA hybrid as substrate, chemical step is represented as irreversible, because reverse reaction, pyrophosphorolysis with formation of a triphosphate is 10000000-times slower than the forward reaction
-
ir
ATP + ATP
P1,P3-bis(5'-adenosyl) triphosphate + ?
show the reaction diagram
-
-
-
?
ATP + ATP
P1,P4-bis(5'-adenosyl) tetraphosphate + diphosphate
show the reaction diagram
-
-
-
?
ATP + clodronate
adenosine 5'-(beta,gamma-dichloromethylenetriphosphate) + ?
show the reaction diagram
-
-
-
-
?
ATP + dimethylallyl diphosphate
dimethylallyl-triphospho(5')adenosine + diphosphate
show the reaction diagram
-
assay contains pyruvate kinase, phosphoenolpyruvate and pyrophosphatase to recover ATP
-
-
ir
ATP + etidronate
?
show the reaction diagram
-
weak activity
-
-
?
ATP + geranyl diphosphate
geranyl-triphospho(5')adenosine + diphosphate
show the reaction diagram
-
assay contains pyruvate kinase, phosphoenolpyruvate and pyrophosphatase to recover ATP
-
-
ir
ATP + geranyl triphosphate
geranyl-tetraphospho(5')adenosine + diphosphate
show the reaction diagram
-
assay contains pyruvate kinase, phosphoenolpyruvate and pyrophosphatase to recover ATP
-
-
ir
ATP + isopentenyl diphosphate
adenosine 5'-isopentenyl triphosphate + diphosphate
show the reaction diagram
-
assay contains pyruvate kinase, phosphoenolpyruvate and pyrophosphatase to recover ATP
-
-
ir
ATP + isopentenyl triphosphate
isopentenyl-tetraphospho(5')adenosine + diphosphate
show the reaction diagram
-
assay contains pyruvate kinase, phosphoenolpyruvate and pyrophosphatase to recover ATP
-
-
ir
ATP + methylenebisphosphonate
adenosine 5'-(beta,gamma-methylenetriphosphate) + ?
show the reaction diagram
-
-
-
-
?
ATP + RNA + 5'-aaCCAGUC
AMP + diphosphate + ?
show the reaction diagram
-
DNA-RNA hybrid
-
?
ATPgammaS + ATP
P1,P4-bis(5'-adenosyl) tetraphosphate + thiodiphosphate
show the reaction diagram
-
-
-
?
dATP + (ribonucleotide)n + (ribonucleotide)m
dAMP + phosphate + (ribonucleotide)n+m
show the reaction diagram
GTP + ATP
P1-(5'-adenosyl),P4-(5'-guanosyl) tetraphosphate + diphosphate
show the reaction diagram
-
-
-
?
ITP + (ribonucleotide)n + (ribonucleotide)m
IMP + phosphate + (ribonucleotide)n+m
show the reaction diagram
-
-
-
-
-
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + (ribonucleotide)n + (ribonucleotide)m
?
show the reaction diagram
ATP + (ribonucleotide)n + (ribonucleotide)m
AMP + diphosphate + (ribonucleotide)n+m
show the reaction diagram
additional information
?
-
-
the group I intron from cyanobacterium Anabaena sp. catalyzes phosphodiester bond formation using a triphosphate on the 5'-terminal nucleotide, much like protein polymerases and engineered ribozymes. In the process, this ribozyme forms a unique circular RNA that incorporates the exogenous guanosine cofactor added during self-splicing
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-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
no detectable activity with Ca2+, Sr2+, Ba2+, Zn2+, Co2+, Cd2+, Pb2+, Co(NH3)6 3+, or spermine
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,N6-Ethenoadenosine 5'-triphosphate
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-
1-amino-4-(3-(aminosulfonyl)anilino)-9,10-dioxo-9,10-dihydro-2-anthracenesulfonic acid
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68.5% residual activity at 0.01 mM, presence of Triton X-100. Molecular dynamics simulations
2',5'-ADP
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poly(A) cyclization
2-naphthoic acid
-
-
2-nitronaphthalene
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-
3-((4-(ethylamino)phenyl)diazenyl)-4,5-dihydroxy-2,7-naphthalenedisulfonic acid
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4.1% residual activity at 0.01 mM, presence of Triton X-100. Molecular dynamics simulations
3-((5-chloro-2-hydroxyphenyl)diazenyl)-4,5-dihydroxy-2,7-naphthalenedisulfonic acid
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8.6% residual activity at 0.01 mM, presence of Triton X-100. Molecular dynamics simulations
4,5-dihydroxy-3-(1-naphthyldiazenyl)-2,7-naphthalenedisulfonic acid
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43.1% residual activity at 0.01 mM, presence of Triton X-100. Molecular dynamics simulations
4,5-dihydroxynaphthalene-2,7-disulfonate
-
-
4-hydroxy-3-[(E)-(5-hydroxy-7-sulfonaphthalen-2-yl)diazenyl]-7-[(1-phenylethenyl)amino]naphthalene-2-sulfonic acid
-
-
6-Chloropurine riboside 5'-triphosphate
-
-
6-hydroxy-5-[(E)-[3-[(E)-(2-hydroxynaphthalen-1-yl)diazenyl]-5-sulfophenyl]diazenyl]naphthalene-2-sulfonic acid
-
-
6-mercaptopurine riboside 5'-triphosphate
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-
7,7'-iminobis(4-hydroxynaphthalene-2-sulfonic acid)
-
-
7-amino-4-hydroxy-3-[(E)-(5-hydroxy-7-sulfonaphthalen-2-yl)diazenyl]naphthalene-2-sulfonic acid
-
-
8-bromoadenosine 5'-triphosphate
-
-
8-Sulfoadenosine 5'-triphosphate
-
-
adenosine 5'-tetraphosphate
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-
adenosine-N1-oxide 5'-triphosphate
-
-
ATP
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above 2.5 mM, complete inhibition at 10 mM. The inhibition can be partially rescued by adding more Mg2+ to the reaction
beta,gamma-imine-ATP
-
-
beta,gamma-methylene-ATP
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-
Co(NH3)63+
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-
gamma-Thio-ATP
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-
Mg2+
-
inhibits above 10 mM, required at lower concentrations
naphthalene-2-sulfonic acid
-
-
P1,P3-di(adenosine-5')triphosphate
-
-
P1,P4-di(adenosine-5')tetraphosphate
-
-
tripolyphosphate
-
-
additional information
-
inhibitors in infectious microorganisms
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
RNA
-
stimulates exchange reaction between ATP and diphosphate
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.24
(2E,6E)-farnesyl diphosphate
-
-
0.07
(2E,6E)-farnesyl triphosphate
-
-
0.009
(ribonucleotide)n+m
-
pH 8.0, 22°C
0.4
5'-adenylated cytidine 5',3'-bisphosphate
-
pH 7.4, 30°C
0.00001
5'-phosphate termini of RNA
-
-
-
0.0002 - 0.04
ATP
0.54
clodronate
-
-
4
diguanosine tetraphosphate
-
pH 7.4, 30°C
0.86
dimethylallyl diphosphate
-
-
4.3
etidronate
-
-
0.29
geranyl diphosphate
-
-
0.23
geranyl triphosphate
-
-
4.9
isopentenyl diphosphate
-
-
3.6
isopentenyl triphosphate
-
-
0.26
methylenebisphosphonate
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
12.1
(2E,6E)-farnesyl diphosphate
Enterobacteria phage T4
-
-
11.6
(2E,6E)-farnesyl triphosphate
Enterobacteria phage T4
-
-
1.66 - 2.94
(ribonucleotide)n+m
0.05 - 6.08
(RNA)n
0.055
ATP
Enterobacteria phage T4
-
pH 7.4, 30°C, ATP as acceptor of AMP, Ap4A synthesis
0.29
clodronate
Enterobacteria phage T4
-
-
3.1
dimethylallyl diphosphate
Enterobacteria phage T4
-
-
0.028
etidronate
Enterobacteria phage T4
-
-
8.6
geranyl diphosphate
Enterobacteria phage T4
-
-
52.2
geranyl triphosphate
Enterobacteria phage T4
-
-
0.017
GTP
Enterobacteria phage T4
-
pH 7.4, 37°C
0.9
isopentenyl diphosphate
Enterobacteria phage T4
-
-
6.7
isopentenyl triphosphate
Enterobacteria phage T4
-
-
0.28
methylenebisphosphonate
Enterobacteria phage T4
-
-
additional information
additional information
Enterobacteria phage T4
-
the kinetic mechanism of single-turnover nick sealing by T4 Rnl2-AMP is explored by using a rapid mix-quench method and the effects of 3'-OH mispairs and base damage lesions on the rates of nick 5'-adenylylation and phosphodiester synthesis is determined. With respect to the sealing of perfectly paired nicks the rates of step 2 catalysis are rapid (9.5-17.9/sec) and similar in magnitude to the step 3 rates (7.9-32/sec). Rnl2 is kinetically sensitive to all 3'-OH base mispairs
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.9
(2E,6E)-farnesyl diphosphate
Enterobacteria phage T4
-
-
81
166
(2E,6E)-farnesyl triphosphate
Enterobacteria phage T4
-
-
27619
2.7
dimethylallyl diphosphate
Enterobacteria phage T4
-
-
157
2.5
geranyl diphosphate
Enterobacteria phage T4
-
-
175
12
geranyl triphosphate
Enterobacteria phage T4
-
-
8877
4.5
isopentenyl diphosphate
Enterobacteria phage T4
-
-
113
24
isopentenyl triphosphate
Enterobacteria phage T4
-
-
8876
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00216
4-hydroxy-3-[(E)-(5-hydroxy-7-sulfonaphthalen-2-yl)diazenyl]-7-[(1-phenylethenyl)amino]naphthalene-2-sulfonic acid
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00159
6-hydroxy-5-[(E)-[3-[(E)-(2-hydroxynaphthalen-1-yl)diazenyl]-5-sulfophenyl]diazenyl]naphthalene-2-sulfonic acid
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00836
7,7'-iminobis(4-hydroxynaphthalene-2-sulfonic acid)
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00153
7-amino-4-hydroxy-3-[(E)-(5-hydroxy-7-sulfonaphthalen-2-yl)diazenyl]naphthalene-2-sulfonic acid
Trypanosoma brucei
-
pH and temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00152
-
-
0.0034
-
with geranyl diphosphate as substrate
0.0037
-
with dimethylallyl diphosphate as substrate
0.004
-
with (2E,6E)-farnesyl diphosphate as substrate
0.0062
-
with isopentenyl diphosphate as substrate
0.016
-
with (2E,6E)-farnesyl triphosphate as substrate
0.0166
-
with geranyl triphosphate as substrate
0.0335
-
with isopentenyl triphosphate as substrate
0.0383
-
5'-phosphate terminus rendered resistant to phosphatase
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 6.5
-
in the presence of Mg2+, the reaction has a pH optimum of 6.0–6.5
6.5
-
adenylyltransferase activity
7.5 - 8
-
MOPS buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9.5
-
activity is virtually nil at pH below 5.0 or above 9.5
5.5 - 8
-
adenylyltransferase activity
6.5 - 8
-
around 70% of the maximum at pH 8.0
7 - 8.5
-
pH 7.0: about 40% of maximal activity, pH 8.5: about 30% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
37
-
assay at
65
-
RNA-adenylylate intermediates AppRNA are predominantly ligated to a circular product
65 - 70
-
-
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35
-
phosphodiester bond formation is suppressed and the majority of the RNA-adenylylate intermediates AppRNA are deadenylylated
50 - 75
-
50°C: about 35% of maximal activity, 75°C: about 60% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Clostridium thermocellum (strain ATCC 27405 / DSM 1237 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372)
Clostridium thermocellum (strain ATCC 27405 / DSM 1237 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372)
Clostridium thermocellum (strain ATCC 27405 / DSM 1237 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
43000
-
x * 43000, SDS-PAGE
47000 - 48200
-
gel filtration, high-speed sedimentation equilibrium centrifugation
48000
-
SDS-PAGE
50000
-
immunoblotting
51300
-
calculated from the putative RNA ligase 1 gene
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 43000, SDS-PAGE
dimer
-
2 * 45000, calculated and sedimentation analysis
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2.6 A crystal structure
-
2.2 A resolution, crystal structure of a complete RNA ligase, Rnl1, in complex with adenosine 5'-(alpha,beta-methylenetriphosphate)
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crystal structure of the ligase domain with AMP bound at the active site, space group P2(1)2(1)2, a = 57.72 A, b = 89.89 A, c = 47.74 A
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hanging-drop vapor diffusion. Crystal structures of: 1. Covalent Rnl2-AMP intermediate, 2. Rnl2 bound to an adenylylated nicked duplex, captured immediately following reaction step 2, 3. Rnl2 at an adenylylated nick in a state poised for step 3. These structures illuminate the stereochemistry of nucleotidyl transfer and reveal how remodeling of active-site contacts and conformational changes propel the ligation reaction forward
-
crystal structure of the catalytic domain of TbREL1 at 1.2 A resolution, in complex with ATP and magnesium.The magnesium ion interacts with the beta and gamma-phosphate groups and is almost perfectly octahedrally coordinated by six phosphate and water oxygen atoms. Sitting-drop, vapor-diffusion with 100 mM HEPES, 100 mM Mg2+ and 35% (w/v) PEG3350 at pH 7.0. The crystal belongs to space group P2(1), with cell dimensions of a = 44.9 A, b = 58.6 A, c = 53.0 A and beta = 100.2°, and one molecule in the asymmetric unit
-
virtual screen of REL1 crystal structure for inhibitors
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
stable for at least 2 h
56
-
2 min, complete inactivation
60 - 65
-
optimal for stability
60 - 90
-
relatively stable at 60°C with a half-life of 10 h, activity increases after incubation, not stable at temperatures. Enzyme starts to lose activity rapidly, no activity at all after 1 h at 80°C, loses all of its activity after 5 min at 90°C
65
-
realatively stable at
80
-
loses all activity above
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
activity is unaffected by at least 3 freeze-thaw cycles
-
extremely unstable, when stored at low ionic strength
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, stable for at least 6 months
-
-70°C, stable for at least 1 year
-
4°C, 32% loss of activity after 2 weeks
-
in liquid nitrogen, at 0.5 mg protein per ml, stable for at least 6 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a complex with 4 activities predicted to catalyze RNA editing: gRNA-directed endonuclease, terminal uridyl transferase, 3' U-specific endonuclease, RNA ligase
-
copurifies with 5'-hydroxyl polynucleotide kinase
-
editing complex
-
histidine-tagged
-
mutant enzymes
-
mutant enzymes, expressed in bacteria
-
produced in Escherichia coli
-
purified via a C-terminal tandem affinity purification tag
-
recombinant
-
recombinant enzyme
recombinant protein
-
using Ni-NTA chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
encoded by T4 gene 24.1, mutants expressed in Escherichia coli
-
enzymatically active Leishmania tarentolae REL1 and REL2 and Trypanosoma brucei REL1 proteins from a Baculovirus expression system. The recombinant proteins can each integrate into an L-complex that has been depleted of the cognate ligase by RNAi. In the case of REL1, the integration of the recombinant enzyme functionally complements the REL1-depleted L-complex in an in vitro editing system
expressed in Escherichia coli
-
expressed in Escherichia coli as a His-tagged fusion protein
expressed in Escherichia coli as a His10-tagged fusion protein
-
expressed in Sf9 insect cells
-
expressed in Trypanosoma brucei lines as an epitope-tagged fusion protein, recombinant p48 also expressed in Escherichia coli
-
expression in Escherichia coli
expression in Escherichia coli BL21goldDE3
-
overexpressed in Escherichia coli BL21 (DE3)
-
RNA ligase gene is inserted into a pET23b expression vector with a C-terminal His6-tag. This vector is transformed into Escherichia coli strain BL21
-
RnlA gene overexpressed in Escherichia coli BL21
-
transformed into Escherichia coli BL21 (DE3)
-
transformed into Escherichia coli BL21(DE3)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
DELTA1-105
-
reacts with ATP to form covalent protein-adenylate adducts,mutant protein retains RNA sealing activity
DELTA1-125
-
reacts with ATP to form covalent protein-adenylate adducts, mutant protein retains RNA sealing activity
DELTA1-135
-
does not reacts with ATP to form covalent protein-adenylate adducts, mutant enzyme is unable to seal RNA strands
E230A
-
mutant is defective in phosphodiester formation at a preadenylylated nick; mutant is dysfunctional in ligase adenylylation
E230D
-
mutation reduces nick sealing activity to 1% of the wild-type level
E230Q
-
mutation reduces nick sealing activity to less than 1% of the wild-type level
E278A
-
mutant enzyme retains adenylyltransferase and RNA ligase activities
E305A
-
mutant is defective in phosphodiester formation at a preadenylylated nick; mutant is dysfunctional in ligase adenylylation
E305D
-
mutation reduces nick sealing activity to 1% of the wild-type level
E305Q
-
mutation reduces nick sealing activity to 7% of the wild-type level
F281A
-
mutant is defective in phosphodiester formation at a preadenylylated nick; mutant is dysfunctional in ligase adenylylation
F281L
-
mutation reduces nick sealing activity to 5% of the wild-type level
G168A
-
mutant is defective in phosphodiester formation at a preadenylylated nick; mutant is dysfunctional in ligase adenylylation
H167A
-
mutant is dysfunctional in ligase adenylylation
H167N
-
mutation reduces nick sealing activity to 7% of the wild-type level
H167Q
-
mutation reduces nick sealing activity to less than 1% of the wild-type level
K165A
-
mutant is inert in enzyme-adenylate formation and nick-joining
K186A
-
mutant is dysfunctional in ligase adenylylation
K186Q
-
mutation reduces nick sealing activity to 1% of the wild-type level
K186R
-
mutation reduces nick sealing activity to 3% of the wild-type level
K326Q
-
mutation reduces nick sealing activity to less than 1% of the wild-type level
K326R
-
mutation reduces nick sealing activity to 3% of the wild-type level
S185N
-
mutation reduces nick sealing activity to less than 1% of the wild-type level
S185T
-
mutation reduces nick sealing activity to 12% of the wild-type level
T163A
-
mutant is dysfunctional in ligase adenylylation
T163S
-
mutation reduces nick sealing activity to 2% of the wild-type level
T163V
-
mutation reduces nick sealing activity to 30% of the wild-type level
D120A
-
alanine-scanning mutagenesis
D135A
-
about 5% of the ligation activity of wild-type enzyme, mutant enzyme displays 4% of the adenylyltransferase activity observed with wild-type enzyme
D244A
-
site-directed mutagenesis
D292A
-
no ligation activity
D292E
-
about 10% of the ligation activity of wild-type enzyme
D308A
-
about 70% of the ligation activity of wild-type enzyme
E139A
-
about as active as wild-type enzyme in RNA ligation, mutant displays near wild-type adenylyltransferase activity
E185A
-
site-directed mutagenesis
E204A
-
substitution mutation introduced into the ORFb y PCR with two-stage overlap extension method
E227A
-
site-directed mutagenesis
E227D
-
site-directed mutagenesis
E227Q
-
site-directed mutagenesis
E295A
-
about as active as wild-type enzyme in RNA ligation
E296A
-
about 10% of the ligation activity of wild-type enzyme
E299A
-
about 80% of the ligation activity of wild-type enzyme
E29A
-
about 90% of the RNA ligation activity of wild-type enzyme, mutant displays near wild-type adenylyltransferase activity
E300A
-
about 50% of the ligation activity of wild-type enzyme
E34A
-
alanine-scanning mutagenesis
E34D
-
substitution mutant, introduced into the rnl2 gene by PCR
E34Q
-
substitution mutant, introduced into the rnl2 gene by PCR
E63A
-
about 80% of the RNA ligation activity of wild-type enzyme, mutant displays near wild-type adenylyltransferase activity
E81A
-
site-directed mutagenesis
E99A
-
alanine-scanning mutagenesis
F119A
-
alanine-scanning mutagenesis
F76A
-
site-directed mutagenesis
F77A
-
site-directed mutagenesis
F77L
-
site-directed mutagenesis
G102A
-
site-directed mutagenesis
G228A
-
site-directed mutagenesis
G55A
-
site-directed mutagenesis
H250A
-
pRNA circularization by the mutant enzyme is 6% of the wild-type rate
H37A
-
substitution mutation introduced into the ORFb y PCR with two-stage overlap extension method
H37D
-
substitution mutation introduced into the ORFb y PCR with two-stage overlap extension method
K107A
-
about as active as wild-type enzyme in RNA ligation, mutant displays near wild-type adenylyltransferase activity
K117A
-
mutation is lethal in vivo
K119A
-
site-directed mutagenesis
K119Q
-
site-directed mutagenesis
K119R
-
site-directed mutagenesis
K189A
-
alanine-scanning mutagenesis
K209A
-
alanine-scanning mutagenesis
K225A
-
substitution mutation introduced into the ORFb y PCR with two-stage overlap extension method
K227A
-
substitution mutation introduced into the ORFb y PCR with two-stage overlap extension method
K240A
-
site-directed mutagenesis
K240Q
-
site-directed mutagenesis
K240R
-
site-directed mutagenesis
K242A
-
site-directed mutagenesis
K273A
-
about as active as wild-type enzyme in RNA ligation
K314A
-
about as active as wild-type enzyme in RNA ligation
K315A
-
about 90% of the ligation activity of wild-type enzyme
K319A
-
about as active as wild-type enzyme in RNA ligation
K35A
-
substitution mutation introduced into the ORFb y PCR with two-stage overlap extension method
K54A
-
about 15% of the RNA ligation activity of wild-type enzyme, mutant displays near wild-type adenylyltransferase activity
K75A
-
site-directed mutagenesis
K75Q
-
site-directed mutagenesis
K75R
-
site-directed mutagenesis
K99A
-
site-directed mutagenesis
L104A
-
L104A strain is temperature sensitive, no growth at 37°C; pRNA circularization by the mutant enzyme is 6% of the wild-type rate
N184A
-
site-directed mutagenesis
N309A
-
about 70% of the ligation activity of wild-type enzyme
N40A
-
alanine-scanning mutagenesis
N40D
-
substitution mutant, introduced into the rnl2 gene by PCR
N40Q
-
substitution mutant, introduced into the rnl2 gene by PCR
N40R
-
substitution mutant, introduced into the rnl2 gene by PCR
N78A
-
site-directed mutagenesis
Q106A
-
about 20% of the RNA ligation activity of wild-type enzyme, mutant displays near wild-type adenylyltransferase activity
R155A
-
about 35% of the ligation activity of wild-type enzyme, mutant enzyme displays 16% of the adenylyltransferase activity observed with wild-type enzyme
R166A
-
site-directed mutagenesis
R182A
-
site-directed mutagenesis
R221A
-
about 60% of the ligation activity of wild-type enzyme, mutant displays near wild-type adenylyltransferase activity
R266A
-
no ligation activity
R266K
-
about 50% of the ligation activity of wild-type enzyme
R266Q
-
no ligation activity
R33A
-
no ligation activity, mutant enzyme displays 3% of the adenylyltransferase activity observed with wild-type enzyme
R54A
-
site-directed mutagenesis
R54K
-
site-directed mutagenesis
R54Q
-
site-directed mutagenesis
R55A
-
alanine-scanning mutagenesis
R71A
-
site-directed mutagenesis
S103A
-
pRNA circularization by the mutant enzyme is 5% of the wild-type rate; S103A strain is temperature sensitive, no growth at 37°C
S118A
-
S118 strain is temperature sensitive, no growth at 37°C
S170A
-
about 5% of the ligation activity of wild-type enzyme, mutant enzyme displays 8% of the adenylyltransferase activity observed with wild-type enzyme
S272A
-
about 50% of the ligation activity of wild-type enzyme
S292N
-
about 25% of the ligation activity of wild-type enzyme
W329A
-
about as active as wild-type enzyme in RNA ligation
Y136A
-
about 95% of the ligation activity of wild-type enzyme, mutant displays near wild-type adenylyltransferase activity
Y5A
-
about 10% of the RNA ligation activity of wild-type enzyme, mutant enzyme displays 39% of the adenylyltransferase activity observed with wild-type enzyme
D425N
-
lethal mutation
D726A
-
lethal mutation
E218A
-
lethal mutation
E326A
-
lethal mutation
E741A
-
lethal mutation
H1060A
-
lethal mutation
H515A
-
lethal mutation
H515N
-
lethal mutation
H515Q
-
lethal mutation
H777N
-
lethal mutation
H777Q
-
lethal mutation
K152A
-
lethal mutation
K541A
-
lethal mutation
K543A
-
lethal mutation
R463A
-
lethal mutation
R463K
-
lethal mutation
R463Q
-
lethal mutation
R511K
-
lethal mutation
R511Q
-
lethal mutation
S701A
-
lethal mutation
T1001A
-
lethal mutation
T675A
-
lethal mutation
T675V
-
lethal mutation
K109R
-
site-directed mutagenesis
K86R
-
site-directed mutagenesis
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
-
regulated repression of the enzyme blocks editing, this repression is lethal in bloodforms of the parasite, indicating that editing is essential in the mammalian stage of the life cycle, the editing complex, which is present in all kinetoplastid parasites, may thus be a chemotherapeutic target
molecular biology
-
RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE); the properties of TS2126 RNA ligase 1 makes it very attractive for processes like adaptor ligation, and single-stranded solid phase gene synthesis
synthesis
Show AA Sequence (906 entries)
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