Information on EC 3.1.22.4 - crossover junction endodeoxyribonuclease

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.1.22.4
-
RECOMMENDED NAME
GeneOntology No.
crossover junction endodeoxyribonuclease
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
endonucleolytic cleavage at a junction such as a reciprocal single-stranded crossover between two homologous DNA duplexes (Holliday junction)
show the reaction diagram
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-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
CAS REGISTRY NUMBER
COMMENTARY hide
99676-43-4
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
Uniprot
Manually annotated by BRENDA team
Bacillus subtilis bacteriophage SPP1
-
-
-
Manually annotated by BRENDA team
strain BG633, BG501, BG651, DK53, DK54, DK55, DK56
-
-
Manually annotated by BRENDA team
strain YB886
-
-
Manually annotated by BRENDA team
Borrelia burgdorferi
-
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
several strains
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-
Manually annotated by BRENDA team
cell lines irs3, irs1, irs1SF and V79
-
-
Manually annotated by BRENDA team
Methanothermobacter thermautotrophicum
-
-
-
Manually annotated by BRENDA team
no activity in mammalia
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3'-flapped junction in DNA + H2O
?
show the reaction diagram
5'-flapped junction in DNA + H2O
?
show the reaction diagram
-
the enzyme shows a robust activity with a specific cleavage site in the 5'-overhang strand exactly one nucleotide 3' of the branch point
-
-
?
D-loop junction in DNA + H2O
?
show the reaction diagram
-
-
-
-
?
DNA + H2O
?
show the reaction diagram
DNA + H2O
hydrolyzed DNA
show the reaction diagram
DNA junction 1 + H2O
?
show the reaction diagram
-
DNA junction 1 substrate assembled from strands b50, h50,r50,x50, r55, b1-27, b28-50
-
-
?
Holliday junction in DNA + H2O
?
show the reaction diagram
Holliday junction X0 + H2O
?
show the reaction diagram
Holliday junctions in DNA + H2O
?
show the reaction diagram
-
-
-
-
?
intact Holliday junction + H2O
?
show the reaction diagram
-
-
-
-
?
nXO12 junction in single-stranded DNA + H2O
?
show the reaction diagram
-
-
-
-
?
pXO12-3' junction in single-stranded DNA + H2O
?
show the reaction diagram
-
-
-
-
?
replication fork-like junction in DNA + H2O
?
show the reaction diagram
-
-
-
-
?
restriction fork in DANN + H2O
?
show the reaction diagram
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GEN1 preferentially cleaves strand 1 exactly at the branch point, whereas SEND1 preferentially cleaves one nucleotide in the 3' direction of the branch point. Cleavage in strand 2 is not detected with either GEN1 or SEND1 indicating a preference for cleavage of the lagging strand matrix
-
-
?
splayed Y junction in DNA + H2O
?
show the reaction diagram
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-
-
-
?
additional information
?
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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
DNA + H2O
?
show the reaction diagram
DNA + H2O
hydrolyzed DNA
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Divalent cations
KCl
-
optimal concentration in reaction buffer: 200 mM
MgCl2
-
required, optimal concentration: 5-10 mM
MnCl2
-
can substitute for MgCl2, less efficient
spermidine
-
-
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
anti-Endo VII antibodies
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Endo X3
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Ca2+
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strong inhibition
Cds1
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the DNA replication checkpoint kinase Cds1 negatively regulates Mus81/Eme1 to preserve genomic integrity when replication is perturbed
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EDTA
-
comple inhibition
KCl
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reduces activity 10fold at a concentration of 200 mM
potassium glutamate
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reduces activity 10fold at a concentration of 200 mM
RuvA
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inhibits cleavage of certain DNA substrates
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WRWYCR
-
inhibits junction cleavage
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
PCNA protein
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Holliday junction-resolving enzyme Hjc interacts physically with PCNA via a canonical C-terminal PIP motif. This interaction stimulates the junction cleavage activity of Hjc in vitro. PCNA is a toroidal protein that acts as a processivity factor for many proteins. In Sulfolobus solfataricus, PCNA is a heterotrimer that can function as a molecular toolbelt, forming simultaneous interactions with up to three partner proteins with related functions in a DNA-processing pathway
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00000102 - 0.0000055
3'-flapped junction in DNA
0.0000012
D-loop junction in DNA
-
-
-
0.000066 - 0.000073
DNA junction 1 substrate
0.00000052
intact Holliday junction
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in 25 mM Tris-HCl, pH 8.0, 1 mM MgCl2, 0.2 mM dithiothreitol, at 37C
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0.0000031
nXO12 junction in single-stranded DNA
-
-
-
0.0000056
pXO12-3' junction in single-stranded DNA
-
-
-
0.0000073
replication fork-like junction in DNA
-
-
-
0.0000304
splayed Y junction in DNA
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-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.12 - 0.97
3'-flapped junction in DNA
0.09
D-loop junction in DNA
Saccharomyces cerevisiae
-
-
-
0.0065 - 0.05
DNA junction 1
0.00014 - 0.02
Holliday junction 3
0.037
intact Holliday junction
Homo sapiens
-
in 25 mM Tris-HCl, pH 8.0, 1 mM MgCl2, 0.2 mM dithiothreitol, at 37C
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0.001
J1T1
Schizosaccharomyces pombe
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-
-
0.00034
J1T2
Schizosaccharomyces pombe
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-
-
0.000006 - 0.0029
junction 1, unconstrained
0.18
nX12 junction in single-stranded DNA
Homo sapiens
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the nX12 substrate has four 25-bp duplex DNA arms, with a 12-bp homologous core that allows branch migration of the junction, and contains a nick at the cross-over point
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1.2
nXO12 junction in single-stranded DNA
Saccharomyces cerevisiae
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-
-
0.32
pXO12-3' junction in single-stranded DNA
Saccharomyces cerevisiae
-
-
-
0.00416
RC1
Escherichia coli
-
-
-
0.000566
RC1(1,3')
Escherichia coli
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-
-
0.0000047
RC1(2,3')
Escherichia coli
-
-
-
1.35
replication fork-like junction in DNA
Saccharomyces cerevisiae
-
-
-
0.26
splayed Y junction in DNA
Saccharomyces cerevisiae
-
-
-
0.0047
X12 junction in single-stranded DNA
Homo sapiens
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like nX12, without the nick at the cross-over point
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
346300
3'-flapped junction in DNA
Homo sapiens
-
in 25 mM Tris-HCl, pH 8.0, 1 mM MgCl2, 0.2 mM dithiothreitol, at 37C
18563
71830
intact Holliday junction
Homo sapiens
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in 25 mM Tris-HCl, pH 8.0, 1 mM MgCl2, 0.2 mM dithiothreitol, at 37C
197715
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
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PCR fragment cleavage assay; plasmid and lambda DNA cleavage assay
7.5 - 8.5
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7.5 - 10
broad optimum
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 10
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less than 50% of maximal activity above and below
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 45
broad optimum
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
16 - 50
activity range, inactive below or above
20 - 40
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less than 50% of maximal activity above and below
45 - 65
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no multiple turnover catalysis at lower temperatures measurable due to the sensitivity of the assay
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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PSNG13, Bloom's syndrome-associated, BLM, helicase-deficient fibroblasts, PSNF5, BLM-complemented fibroblasts
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
wild type enzyme and truncated enzyme forms DELTA1-35 and DELTA1-35/CDELTA15
-
Manually annotated by BRENDA team
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1)
Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
18000
-
Endo X3, SDS-PAGE
18600
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calculation from amino acid sequence
23000
-
Endo X3, sedimentation in a sucrose density gradient
23950
-
MALDI-TOF MS
24000
-
SDS-PAGE
25000
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gel-filtration
33000
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gel filtration
38000
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SDS-PAGE, enzyme has a globular shape
43000
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Endo X3, gel filtration
53900
-
predicted molecular mass of the BpuJI subunit
54000
Bacillus subtilis bacteriophage SPP1
-
gel filtration
73600
-
predicted molecular weight for His10-FLAG-Mus81, verified by PAGE
75500
-
MUS81 with N-terminal His-tag, gel filtration
82000
-
determined by gel filtration using a 3 microM EcME solution, heterodimer
106400
-
predicted molecular weight for GST-Mms4, verified by PAGE
109000
-
homodimer, determined by sedimentation equilibrium experiments using an analytical ultracentrifuge
140000
-
determined by gel filtration using a 88 microM EcME solution, heterotetramer
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterodimer
homodimer
monomer
octamer
-
active enzyme, in the presence of Mg2+, SDS-PAGE
tetramer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
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-
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
prismatic crystals obtained by sparse-matrix screening and sitting-drop vapour-diffusion, plate-like crystals by counter-diffusion in X-ray capillary
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at 17C, hanging drop method, in solution having a mushroom-like appearance, where the cap is formed by the main beta-sheet and flanking helices, structure of RecU superimposes best with PfHjc and archaeal HJR, the next best matches to RecU are a series of type II restriction endonucleases including EcoRV and Pvull
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resolving the enzyme bound to DNA junctions, X-ray diffraction structure determination and analysis
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hanging-drop method, 2.1 A resolution
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the structures of RusA-D70N and RusA-D70N-DNA complex are determined at resolutions of 1.2 A and 3.1 A, respectively
three distinct crystal forms, form I: triclinic space group P1, resolution: 4 A, form II: not grown to a size sufficient for X-ray diffraction, form III: monoclinic space group P2-1, resolution: 2.5 A
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improvements in crystallization conditions result in an increased resolution of the native enzyme crystal's diffraction data set of 1.4 A
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the structure of Hjc is determined by using multiple-wavelength anomalous dispersion to a resolution of 2.15A
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resolving enzymes bound to DNA junctions, X-ray diffraction structure determination and analysis
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microbatch method with a silicone oil overlay, crystal structure at 2.0 A resolution
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selenomethionine-containing enzyme crystallized by microbatch method with a silicone oil overlay, hexagonal form of the enzyme
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sitting drop method, Mosquito nanolitre-drop crystallization robot, X-ray data collection
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crystal structure of Sulfolobus Hjc, determined by using multiple-wavelength anomalous dispersion, to a resolution of 2.15 A
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hanging-drop method at 20C
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hanging-drop method, 2.4 A resolution
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hanging drop vapor diffusion method, using 35% (w/v) polyethylene glycol 3350, 0.2M Li2SO4 and 0.1M Tris-HCl (pH 8.5)
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in complex with a synthetic Holliday junction, using 0.1 M citric acid (pH 3.5) and 1.5 M NaCl in 0.3-0.5M ammonium phosphate
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 37
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MUS81-EME2 is more stable at 30C than 37C, but remains active at 37C up to 30 min of incubation
95
-
for 2 min followed by transfer on ice, inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the tetrameric RuvA-Holliday junction complex II is stable in the presence of 750 mM NaCl, whereas the same amount of salt induces more than 80% dissociation of the octameric complex I
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 0.6 ml enzyme solution were diluted with 0.6 ml glycerol
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-20C, fraction VI of the Endo X3 purification can be stored after dialysis against buffer E
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-20C, stable for about 2 weeks
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-20C, stable for at least 8 months without measurable loss of activity
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-80C, Tris-HCl buffer, 50% glycerol
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-80C, Tris-HCl buffer, pH 8, 50% glycerol
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on ice, fraction VI of the Endo X3 purification can be stored for over 1 month, should not be frozen
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
1873fold
-
99% purified, SDS-PAGE and Edman degradation
-
about 95% pure; YDC2
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affinity chromatography on amylose resin and gel filtration of MBP-A22
-
affinity-purified
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amylose and heparin agarose chromatography of MBP-Rap, Rap29K, Rap-S and Rap17K
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carboxy-terminal His-tagged GEN1 is purified using HisTrap column chromatography, heparin column chromatography, ssDNA column chromatography, and MonoS column chromatography
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CCE1, 22fold to near homogeneity
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DEAE-cellulose column chromatography, ammonium sulfate fractionation, Superdex-200 gel filtration, phosphocellulose column chromatography, and hydroxyapatite Bio-Gel HTP column chromatography
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EcME is purified to apparent homogeneity through affinity chromatography, ion exchange, and gel filtration steps
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Endo X3, more than 1325fold, purification beyond fraction V requires addition of pure protein like bovine serum albumin to a final concentration of 0.5 mg/ml
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gel filtration of MBP-RuvC
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gel filtration on a Hi-Load (1.6 x 60 cm) Superdex 200 column, 90% purified by SDS-PAGE
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gel filtration on Superose 12 column, > 98% purified
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gel filtration, Superdex 75 column, greater than 98% purified
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Hi-trap heparin-Sepharose column chromatography
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highly purified
-
highly purified in bacteria, partially purified from the cognate host
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highly purified, 99% pure; RuvC
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hydroxyapatite column chromatography, MonoQ column chromatography, and Superdex 75 gel filtration
Bacillus subtilis bacteriophage SPP1
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Ni2+-NTA column chromatography and heparin column chromatography
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nickel affinity column chromatography and Mono-S column chromatography
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overexpressed GST-Mms4/His10-Flag-Mus81 is purified by sequential affinity chromatography from Saccharomyces cerevisiae cells
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partial purification
-
partial; partial
phosphocellulose and DNA agarose chromatography, FPLC-gel filtration, 99% purified
-
produced in Escherichia coli
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purification of recombinant fusion protein MBP-YDC2
-
purified by dual affinity chromatography
-
purified to homogeneity
using heparin-Sepharose, blue-Sepharose and AH-Sepharose columns
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
amino acids 260-551 for Mus81 and amino acids 244-571 for Eme1 are cloned for expression in Escherichia coli, the complex is named EcME
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carboxy-terminal His-tagged GEN1 is expressed in Escherichia coli
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cloned into the pDuetMxe vector for overexpression in Escherichia coli BL21DE3pLysS cells
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cloning in Escherichia coli strain NEB 5 alpha, and expression in strain NEB Turbo and NEB T7 Express
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cloning of MUS81 and MMS4 into GAL1/10 divergent promotor, 2 micro-based overexpression vector, based on pJN58
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construction of a MBP-YDC2 fusion protein
-
expressed in Escherichia coli
expressed in Escherichia coli BL21 (DE3) pLysS cells
Bacillus subtilis bacteriophage SPP1
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli BL21(DE3) [pLysS] cells
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expressed in Escherichia coli BL21(DE3)[pLysS] cells
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expressed in Escherichia coli BL21-Codon plus (DE3) RIL cells
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expressed in Escherichia coli strain Origami (DE3)pLysS
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expressed in Saccharomyces cerevisiae strain W303
-
expression in Escherichia coli
expression in Escherichia coli Bl21(DE3) strains Bl21-Al and Bl21 pLysS
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expression in Escherichia coli BL21(DE3)(pLysS)
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expression in Escherichia coli containing pMALc2
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expression in Escherichia coli ruvABC-deficient strain
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expression in Escherichia coli strain BL2 (DE3) Codon Plus RIL
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expression in Escherichia coli strain BL21(DE3) Codon Plus RIL
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expression in Escherichia coli using vector pET19b-Hje with a native N-terminal
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expression of covalently linked endonuclease VII dimers in Escherichia coli BL21
expression of MBP-RuvC in Escherichia coli strain ER2508
expression of mutant enzymes in Escherichia coli
-
expression of wild-type and mutant enzymes D30N and E81Q
-
for expression in Escherichia coli
for expression in Escherichia coli ER2267 cells
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gene gen-1, DNA and amino acid sequence determination and analysis. GEN-1 assignment and phylogenetic relationships
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gene MG352, expression of wild-type and mutant enzymes as MBP fusion protein sin Escherichia coli strain BL21(DE3)
-
gene RecUMge, DNA and amino acid sequence determination and analysis
HJE gene cloned from genomic DNA into the pET19b Escherichia coli expression vector and the recombinant protein is expressed at high levels
-
in Escherichia coli strain BL21(DE3)(pLysS) containing the recU overexpression plasmid pCB210
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overexpression in a baculovirus system
-
overexpression in Escherichia coli BL21
overexpression in Escherichia coli BL21(DE3)(pLysS) and XL1-Blue
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YEN1 coding sequence amplified from BY4741 genomic DNA and cloned into pDONR221. Further subcloning of the wild-type or mutant into pYES-DEST52 or pAG416GPD-ccdB-HA to generate pYES-DEST52-YEN1-V5-6xHis, pAG416GPD-YEN1-3xHA or pAG416GPDYEN1E193A/E195A-3xHA. Coding sequence of Yen1-V5-6xHis amplified from pYES-DEST52-YEN1-V5-6xHis and subcloned into the BamHI and HindIII sites of p416ADH to generate p416ADH-YEN1-V5-6xHis
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F81A
-
naturally occuring mutant, that is sensitive to DNA-damaging agents as a null recU strain, a severely impaired enzyme. The mutant poorly recognizes and distorts Holliday junctions. At high concentrations, RecUF81A binds to Holliday junctions but fails to cleave them. RecUF81A does not inhibit RecA dATPase and strand-exchange activities and it loses structural selectivity for X-shaped structures. Phenotype, detailed overview
K56A
-
mutant with a putative separation-of-function phenotype, the mutant is about 5times less active than the wild type enzyme in cleavage of Holliday junctions, fails to inhibit the dATPase activity of RecA because it does not bind ssDNA
R71A
-
mutant with a putative separation-of-function phenotype, fails to inhibit the dATPase activity of RecA because it does not bind ssDNA
Y80A
-
naturally occuring mutant showing an an intermediate phenotype. The mutant poorly recognizes and distorts Holliday junctions, and cleaves them with low efficiency. RecUY80A does not inhibit RecA dATPase and strand-exchange activities and it loses structural selectivity for X-shaped structures. Phenotype, detailed overview
D292N
-
1.2fold decrease in binding of four-way DNA junction, 80fold decrease in catalytic activity
D293N
-
1.2fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
D294N
-
1.2fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
E145Q
-
0.85fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
E231A
-
21fold decrease in binding of four-way DNA junction, 4fold decrease in catalytic activity
F79A
-
70fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
K291A
-
130fold decrease in binding of four-way DNA junction, 47fold decrease in catalytic activity; 30fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
K291R
-
130fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
Q147A
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7.5fold decrease in binding of four-way DNA junction, 100fold decrease in catalytic activity
R146A
-
70fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
R150A
-
18fold decrease in binding of four-way DNA junction, 70fold decrease in catalytic activity
R231K
-
330fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
D70N
catalytically inactive mutant
E68G/H136R
-
the mutation affects octamer formation, DNA binding, and the stimulation of RuvB helicase activity
H29R/E40G/E68G/K129E/F140S/S177G/D184N
-
the mutation affects octamer formation, DNA binding and the stimulation of RuvB helicase activity
H29R/E40G/Q58R/K129E/F140S/S177G/D184N
-
the mutation affects octamer formation, DNA binding and the stimulation of RuvB helicase activity
K76Q
-
failure of the mutant enzyme to promote DNA repair
K76R
-
failure of the mutant enzyme to promote DNA repair
N73A
-
mutant enzyme with 20% of the wild-type activity at 50 nM protein concentration
N79D/N100D
-
the mutation affects octamer formation, DNA binding and the stimulation of RuvB helicase activity
R69A
-
failure of the mutant enzyme to promote DNA repair
R69Q
-
failure of the mutant enzyme to promote DNA repair
D132A
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D132C
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D135A
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D135C
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D135N
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D55A
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D7A
-
site-directed mutagenesis, inactive mutant
D7C
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D7N
-
site-directed mutagenesis, inactive mutant, the active-site mutant resolvase holds the arms in a more planar arrangement in EDTA, Ca2+, or Mg2+ conditions
E33A
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
E60A
-
site-directed mutagenesis, inactive mutant
E60C
-
site-directed mutagenesis, mutant with reduced activity, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
E60N
-
site-directed mutagenesis, mutant with reduced activity, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
E60Q
-
site-directed mutagenesis, inactive mutant
K102A
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
K102R
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
R13M
-
mutant is used to include a second methionine in addition to Met-56
D307A
-
D307 is critical for catalysis
D338A/D339A
-
mutant, lacking potential catalytic residues
Y141A/L142A/N143A/V148A
-
the mutations completely abrogate GEN1 centrosome accumulation
D8N
-
mutant defective in one of four conserved residues known to comprise the catalytic site, slightly improved binding to DNA but unable to cleave it
D112A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding and cleaving activities
D57A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but no cleaving activities
D68A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but no cleaving activities
E11A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but no cleaving activities
E70A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but no cleaving activities
F103A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
F108A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
F69A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
F79A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
G100A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
G60A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
G64A
-
site-directed mutagenesis, the mutant shows slightly reduced Holliday junction binding but unaltered cleaving activities compared to the wild-type enzyme
G7A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but reduced cleaving activities compared to the wild-type enzyme
H87A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but reduced cleaving activities compared to the wild-type enzyme
H91A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
K31A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
K72A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
K76A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
L10A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding and cleaving activities
L122A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but reduced cleaving activities compared to the wild-type enzyme
L92A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
M8A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding and cleaving activities
N15A
-
site-directed mutagenesis, the mutant shows slightly reduced Holliday junction binding and cleaving activities
N5A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding and cleaving activities
Q88A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but no cleaving activities
S54A
-
site-directed mutagenesis, the mutant shows slightly reduced Holliday junction binding and cleaving activities
T74A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
V46A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but reduced cleaving activities compared to the wild-type enzyme
Y58A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
Y62A
-
site-directed mutagenesis, the mutant shows slightly reduced Holliday junction binding but unaltered cleaving activities compared to the wild-type enzyme
Y66A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
G64A
-
site-directed mutagenesis, the mutant shows slightly reduced Holliday junction binding but unaltered cleaving activities compared to the wild-type enzyme
-
H91A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
-
K72A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
-
K76A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
-
DELTA1-5
-
mutation causes a considerable decrease in Hjc-Holliday junction complex formation and cleavage activity
E110A
-
mutant enzyme is as active as the wild-type enzyme
E11A
-
mutant enzyme is as active as the wild-type enzyme
E46A
-
no cleavage of Holliday junction
E9A
-
no cleavage of Holliday junction
F21A
-
mutant enzyme is as active as the wild-type enzyme
F68A
-
no cleavage of Holliday junction
F72A
-
no cleavage of Holliday junction. Mutant enzyme exists as monomer more frequently in solution than as dimer
F89A
-
mutant enzyme is as active as the wild-type enzyme
K30A/K31A
-
mutant enzyme retains proper binding ability to the Holliday junction, little or almost no cleavage activity
K48A
-
no cleavage of Holliday junction. Mutant enzyme exists as monomer more frequently in solution than as dimer
K51A/K52A
-
mutant enzyme retains proper binding ability to the Holliday junction, weak cleavage activity
K81A
-
no cleavage of Holliday junction
R10A
-
no cleavage of Holliday junction
R25A
-
no cleavage of Holliday junction
R3A/K4A
-
mutation reduces the activity by 20fold as compared with the wild-type enzyme. The binding to the Holliday junction is substantially lowered
Y56A
-
some decrease in activity
D414A/D415A
-
mutations introduce a diagnostic NheI restriction site
E193A/E195A
-
catalytically inactive
DELTA1-35
-
mutant enzyme is not able to resolve the synthetic four-way junction X12
DELTA1-35/CDELTA15
-
mutant enzyme is not able to resolve the synthetic four-way junction X12
E226N
-
inactive mutant of YDC2
T239A
-
mutant lacking Cds1-dependent regulation
S30A
-
serine 30 on a flexible loop is catalytically essential, mutants show a decrease in catalytic rate of 3-4 orders of magnitude
S30C
-
serine 30 on a flexible loop is catalytically essential, mutants show a decrease in catalytic rate of 3-4 orders of magnitude
S30T
-
has a slightly higher activity than mutants S30A and S30C but is still severely compromised
D146N
-
inactive
E70Q
-
inactive
F73A
-
the mutant shows 50% reduced activity compared to the wild type enzyme
F74A
-
the mutant shows 30% reduced activity compared to the wild type enzyme
H143D
-
the mutant shows increased activity compared to the wild type enzyme
L80C
-
inactive
M108C
-
the mutant shows reduced activity compared to the wild type enzyme
P40C
-
the mutant shows reduced activity compared to the wild type enzyme
Q77C
-
inactive
R140C
-
the mutant shows reduced activity compared to the wild type enzyme
R76C
-
inactive
T11C
-
the mutant shows reduced activity compared to the wild type enzyme
Y75A/H143D
-
the mutant shows increased activity compared to the wild type enzyme
D146N
-
inactive
-
E70Q
-
inactive
-
F73A
-
the mutant shows 50% reduced activity compared to the wild type enzyme
-
F74A
-
the mutant shows 30% reduced activity compared to the wild type enzyme
-
H143D
-
the mutant shows increased activity compared to the wild type enzyme
-
T11C
-
the mutant shows reduced activity compared to the wild type enzyme
-
Y75A
-
the mutant shows about 1.9fold increase of activity compared to the wild type enzyme; the mutant shows markedly increased activity compared to the wild type
-
Y75A/H143D
-
the mutant shows increased activity compared to the wild type enzyme
-
D151A
-
mutant with amino acid active site substitution
D151N
-
mutant with amino acid active site substitution
D152A
-
mutant with amino acid active site substitution
D152N
-
mutant with amino acid active site substitution
D155A
-
mutant with amino acid active site substitution
D30A
-
mutant with amino acid active site substitution
D81Q
-
mutation eliminates catalytic activity without affecting specific DNA binding
E81A
-
mutant with amino acid active site substitution
E81Q
-
mutant with amino acid active site substitution
K124A
-
mutant with amino acid active site substitution
R121A
-
mutant with amino acid active site substitution
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
Show AA Sequence (10304 entries)
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