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Information on EC 3.1.11.6 - exodeoxyribonuclease VII and Organism(s) Escherichia coli and UniProt Accession P04994

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Escherichia coli
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Word Map
The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Reaction Schemes
Exonucleolytic cleavage in either 5'- to 3'- or 3'- to 5'-direction to yield nucleoside 5'-phosphates
Synonyms
exonuclease vii, exovii, escherichia coli exonuclease vii, exodeoxyribonuclease vii, e. coli exonuclease vii, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E. coli exonuclease VII
-
-
-
-
endodeoxyribonuclease VII
-
-
-
-
Escherichia coli exonuclease VII
-
-
-
-
exonuclease VII
ExoVII
nuclease, exodeoxyribo-, VII
-
-
-
-
XseA
-
large subunit of exonuclease VII
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
52933-20-7
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
single-stranded DNA + H2O
hydrolyzed single-stranded DNA
show the reaction diagram
-
-
?
thymine dimers + H2O
hydrolyzed single-stranded DNA
show the reaction diagram
possible role in methyl-directed mismatch repair system
-
?
duplex DNA containing single-stranded termini + H2O
hydrolyzed single-stranded DNA
show the reaction diagram
msDNA Ec78 + H2O
?
show the reaction diagram
exonucleolytic cleavage
-
-
?
msDNA Ec83 + H2O
?
show the reaction diagram
exonucleolytic cleavage
-
-
?
polydeoxyribonucleotides + H2O
hydrolyzed single-stranded DNA
show the reaction diagram
-
-
-
?
single-stranded DNA + H2O
?
show the reaction diagram
-
releases oligonucleotide products
-
-
?
single-stranded DNA + H2O
hydrolyzed single-stranded DNA
show the reaction diagram
thymine dimers + H2O
hydrolyzed single-stranded DNA
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
single-stranded DNA + H2O
hydrolyzed single-stranded DNA
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
-
25% activation
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
N-ethylmaleimide
-
47% inhibition in absence of mercaptoethanol
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
required for 100% activity
phosphate
sulfate
-
stimulates
additional information
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.8 - 7.9
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
xseA
Uniprot
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
physiological function
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
105000
-
heterodimer, 1 * 54000, 4 * 105000, large subunit: xseA gene, small subunit: xseB gene, SDS-PAGE
51820
-
large subunit, amino acid sequence
54000
-
heterodimer, 1 * 54000, 4 * 105000, large subunit: xseA gene, small subunit: xseB gene, SDS-PAGE
88000
-
gel fltration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
exonuclease VII of Escherichia coli is composed of two different subunits, the large subunit, XseA, and the small subunit, XseB
heterodimer
enzyme ExoVII consists of two subunits that are encoded by xseA and xseB. Gene xseA encodes the catalytically active subunit, while xseB encodes for a smaller subunit which is likely to regulate the activity of ExoVII
pentamer
-
heterodimer, 1 * 54000, 4 * 105000, large subunit: xseA gene, small subunit: xseB gene, SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
structural analysis using bioinformatics. Large subunit XseA consists of four domains: an N-terminal OB-fold domain, a middle putatively catalytic domain, a coiled-coil domain and a short C-terminal segment. The OB-fold domain is responsible for DNA binding, the coiled-coil domain is involved in binding multiple copies of the XseB subunit and residues D155, R205, H238 and D241 of the middle domain are important for the catalytic activity but not for DNA binding
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A188T
site-directed mutagenesis of a highly conserved residue. The A188T mutant shows a msDNA cleavage indistinguishable from that of the wild-type, and lethality is reduced. About 10times more colonies are observed from the A188T mutant than from the wild-type
D155A
-
mutant of large subunit XseA, complete loss of catalytic activity
D155N
site-directed mutagenesis of a highly conserved residue. The D155N mutant loses the ability to cleave msDNA and displays little lethality
D241A
-
mutant of large subunit XseA, complete loss of catalytic activity
DELTA397-456
-
deletion mutant of large subunit XseA, complete loss of catalytic activity
F63A
-
mutant of large subunit XseA, decrease in DNA-binding activity
G237R
site-directed mutagenesis of a residue in the glycine-rich motif. No msDNA cleavage is found for the G237R mutant but a low level of cell killing is still observed. The culture viability drops to about 30% after a 2 h induction
H238A
-
mutant of large subunit XseA, complete loss of catalytic activity
L88R
site-directed mutagenesis
Q96A
-
mutant of large subunit XseA, shows only mild effects
R205A
-
mutant of large subunit XseA, complete loss of catalytic activity
R64E/R68E/R69E
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mutant of large subunit XseA, decrease in DNA-binding activity
V19A
site-directed mutagenesis of a highly conserved residue
W31R
site-directed mutagenesis of a highly conserved residue
additional information
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.3 - 7.8
-
50% loss of activity compared to pH 7.8
134037
7.8 - 8.6
-
50% loss of activity compared to pH 7.8
134037
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
half-life: 90 min
45
-
half-life: 45 min
60
-
half-life: 7 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
freezing and thawing causes loss of activity
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 20 mM Tris-HCl, pH 8, 10 mM mercaptoethanol, 0.1 mM EDTA, 0.2 M NaCl, 50% glycerol, 18 months, 10% loss of activity
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
7500fold to 87% homogeneity
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
genes xseA and xseB, overexpression of small subunit-encoding gene xseB in Escherichia coli, N- and C-terminal ends of large subunit-encoding xseA are separately amplified, cloned independently into a T-vector, and then joined together in pCC1fos, a vector with a very low copy number
large subunit
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strain deficient in enzymic activity plus deficiency in RecJ, ExoI and ExoX exonuclease activities as well as a strain deficient in RecJ, ExoVII and ExoI exonucleases grow poorly in presence of 2-aminopurine. Exposure to 2-aminopurine results in filament formation. The quadruple mutant is cold-sensitive with a severe growth defect at 30°C
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strain deficient in enzymic activity plus deficiency in RecJ, ExoI and ExoX exonuclease activities displays a 7fold increase in mutation rate, the availabilty of any one of thr nucleases is enough to support full mismatch correction
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
additional information
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Chase, J.W.; Richardson, C.C.
Exonuclease VII of Escherichia coli
Basic Life Sci.
5A
225-234
1975
Escherichia coli
Manually annotated by BRENDA team
Chase, J.W.; Vales, L.D.
Exonuclease VII of E. coli
Gene Amplif. Anal.
2
147-168
1981
Escherichia coli
Manually annotated by BRENDA team
Chase, J.W.; Richardson, C.C.
Exonuclease VII of Escherichia coli. Purification and properties
J. Biol. Chem.
249
4545-4552
1974
Escherichia coli
Manually annotated by BRENDA team
Chase, J.W.; Richardson, C.C.
Exonuclease VII of Escherichia coli. Mechanism of action
J. Biol. Chem.
249
4553-4561
1974
Escherichia coli
Manually annotated by BRENDA team
Chase, J.W.; Masker, W.E.
Deoxyribonucleic acid repair in Escherichia coli mutants deficient in the 5'-3' exonuclease activity of deoxyribonucleic acid polymerase I and exonuclease VII
J. Bacteriol.
130
667-675
1977
Escherichia coli
Manually annotated by BRENDA team
Hutton, J.R.; Thomas, C.A.
The assay and isolation of DNA rings using an ATP-dependent endonuclease
Biochemistry
14
1432-1436
1975
Escherichia coli
Manually annotated by BRENDA team
Chase, J.W.; Masker, W.E.; Murphy, J.B.
Pyrimidine dimer excision in Escherichia coli strains deficient in exonucleases V and VII and in the 5' leads to 3' exonuclease of DNA polymerase I
J. Bacteriol.
137
234-242
1979
Escherichia coli
Manually annotated by BRENDA team
Vales, L.D.; Rabin, B.A.; Chase, J.W.
Subunit structure of Escherichia coli exonuclease VII
J. Biol. Chem.
257
8799-8805
1982
Escherichia coli
Manually annotated by BRENDA team
Schultz, R.A.; Friedberg, E.C.; Moses, R.E.; Rupp, W.D.; Sancar, A.; Sharma, S.
Use of an Escherichia coli mutant strain permits measurement of single-stranded apurinic-apyrimidinic endonuclease in crude extracts: studies with untransformed cells and cells transformed with plasmids containing the uvrC gene
J. Bacteriol.
154
1459-1461
1983
Escherichia coli
Manually annotated by BRENDA team
Vales, L.D.; Rabin, B.A.; Chase, J.W.
Isolation and preliminary characterization of Escherichia coli mutants deficient in exonuclease VII
J. Bacteriol.
155
1116-1122
1983
Escherichia coli, Escherichia coli xseB
Manually annotated by BRENDA team
Chase, J.W.; Rabin, B.A.; Murphy, J.B.; Stone, K.L.; Williams, K.R.
Escherichia coli exonuclease VII. Cloning and sequencing of the gene encoding the large subunit (xseA)
J. Biol. Chem.
261
14929-14935
1986
Escherichia coli
Manually annotated by BRENDA team
Chassagne, C.; Schwartz, K.
Mapping of mRNA isoforms with an oligonucleotide probe: exonuclease VII compared with endonucleases
Nucleic Acids Res.
20
3256
1992
Escherichia coli
Manually annotated by BRENDA team
Harris, R.S.; Ross, K.J.; Lombardo, M.J.; Rosenberg, S.M.
Mismatch repair in Escherichia coli cells lacking single-strand exonucleases ExoI, ExoVII, and RecJ
J. Bacteriol.
180
989-993
1998
Escherichia coli (P04994), Escherichia coli
Manually annotated by BRENDA team
Viswanathan, M.; Lovett, S.T.
Single-strand DNA-specific exonucleases in Escherichia coli. Roles in repair and mutation avoidance
Genetics
149
7-16
1998
Escherichia coli
Manually annotated by BRENDA team
Viswanathan, M.; Burdett, V.; Baitinger, C.; Modrich, P.; Lovett, S.T.
Redundant exonuclease involvement in Escherichia coli methyl-directed mismatch repair
J. Biol. Chem.
276
31053-31058
2001
Escherichia coli
Manually annotated by BRENDA team
Burdett, V.; Baitinger, C.; Viswanathan, M.; Lovett, S.T.; Modrich, P.
In vivo requirement for RecJ, ExoVII, ExoI, and ExoX in methyl-directed mismatch repair
Proc. Natl. Acad. Sci. USA
98
6765-6770
2001
Escherichia coli
Manually annotated by BRENDA team
Dermic, D.; Zahradka, D.; Petranovic, M.
Exonuclease requirements for recombination of lambda phage in recD mutants of Escherichia coli
Genetics
173
2399-2402
2006
Escherichia coli
Manually annotated by BRENDA team
Thoms, B.; Borchers, I.; Wackernagel, W.
Effects of single-strand DNases ExoI, RecJ, ExoVII, and SbcCD on homologous recombination of recBCD+ strains of Escherichia coli and roles of SbcB15 and XonA2 ExoI mutant enzymes
J. Bacteriol.
190
179-192
2008
Escherichia coli
Manually annotated by BRENDA team
Dutra, B.E.; Sutera, V.A.; Lovett, S.T.
RecA-independent recombination is efficient but limited by exonucleases
Proc. Natl. Acad. Sci. USA
104
216-221
2007
Escherichia coli, Escherichia coli AB1157
Manually annotated by BRENDA team
Serment-Guerrero, J.; Brena-Valle, M.; Espinosa-Aguirre, J.J.
In vivo role of Escherichia coli single-strand exonucleases in SOS induction by gamma radiation
Mutagenesis
23
317-323
2008
Escherichia coli, Escherichia coli BW25113
Manually annotated by BRENDA team
Larrea, A.A.; Pedroso, I.M.; Malhotra, A.; Myers, R.S.
Identification of two conserved aspartic acid residues required for DNA digestion by a novel thermophilic Exonuclease VII in Thermotoga maritima
Nucleic Acids Res.
36
5992-6003
2008
Escherichia coli, Thermotoga maritima
Manually annotated by BRENDA team
Noothi, S.K.; Minda, R.; Rao, B.J.
MutS and UvrD proteins stimulate exonuclease action: insights into exonuclease-mediated strand repair
Biochemistry
48
7787-7793
2009
Escherichia coli
Manually annotated by BRENDA team
Rudolph, C.J.; Mahdi, A.A.; Upton, A.L.; Lloyd, R.G.
RecG protein and single-strand DNA exonucleases avoid cell lethality associated with PriA helicase activity in Escherichia coli
Genetics
186
473-492
2010
Escherichia coli, Escherichia coli K-12, MG1655
Manually annotated by BRENDA team
Repar, J.; Briski, N.; Buljubasic, M.; Zahradka, K.; Zahradka, D.
Exonuclease VII is involved in "reckless" DNA degradation in UV-irradiated Escherichia coli
Mutat. Res.
750
96-104
2013
Escherichia coli
Manually annotated by BRENDA team
Poleszak, K.; Kaminska, K.H.; Dunin-Horkawicz, S.; Lupas, A.; Skowronek, K.J.; Bujnicki, J.M.
Delineation of structural domains and identification of functionally important residues in DNA repair enzyme exonuclease VII
Nucleic Acids Res.
40
8163-8174
2012
Escherichia coli
Manually annotated by BRENDA team
Jung, H.; Liang, J.; Jung, Y.; Lim, D.
Characterization of cell death in Escherichia coli mediated by XseA, a large subunit of exonuclease VII
J. Microbiol.
53
820-828
2015
Escherichia coli (P04994 AND P0A8G9), Escherichia coli
Manually annotated by BRENDA team
Midgley-Smith, S.L.; Dimude, J.U.; Rudolph, C.J.
A role for 3' exonucleases at the final stages of chromosome duplication in Escherichia coli
Nucleic Acids Res.
47
1847-1860
2019
Escherichia coli (P04994 AND P0A8G9), Escherichia coli
Manually annotated by BRENDA team