Information on EC 3.1.21.5 - type III site-specific deoxyribonuclease

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The expected taxonomic range for this enzyme is: Bacteria, Archaea

EC NUMBER
COMMENTARY
3.1.21.5
-
RECOMMENDED NAME
GeneOntology No.
type III site-specific deoxyribonuclease
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
endonucleolytic cleavage of DNA to give specific double-stranded fragments with terminal 5'-phosphates
show the reaction diagram
-
-
-
-
endonucleolytic cleavage of DNA to give specific double-stranded fragments with terminal 5'-phosphates
show the reaction diagram
mechanism
-
endonucleolytic cleavage of DNA to give specific double-stranded fragments with terminal 5'-phosphates
show the reaction diagram
mechanism
prophage P1
-
endonucleolytic cleavage of DNA to give specific double-stranded fragments with terminal 5'-phosphates
show the reaction diagram
mechanism, formation of loop structures and DNA translocation
-
endonucleolytic cleavage of DNA to give specific double-stranded fragments with terminal 5'-phosphates
show the reaction diagram
mechanism, model of DNA translocation and cleavage, overview
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of phosphoric ester
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
ATP-dependent type III restriction endonuclease
-
-
BsaHI
Geobacillus stearothermophilus CPW11
B0LX59
-
-
EC 3.1.23
-
-
-
-
EC 3.1.24
-
-
-
-
PspGI
-
recognizes the sequence CCWGG where W is A or T and cuts DNA before the first C in the cognate sequence
PstII
Providencia stuartii 164
-
-
-
R.EcoP15I
-
-
restriction endonuclease PstII
-
-
restriction endonuclease PstII
Providencia stuartii 164
-
-
-
restriction-modification system
-
-
-
-
type III DNA restriction/modification enzyme
-
-
type III R/M enzyme
-
-
type III RE
-
-
type III restriction endonuclease
-
-
type III restriction enzyme
-
-
-
-
type III restriction enzyme
-
-
type III RM system
Q5F957
-
type III testriction-modification enzyme
-
-
type III-like restriction endonuclease
-
-
additional information
-
a complete listing of all these enzymes and their recognition sites has been produced by R.J. Roberts, this list is updated annually
CAS REGISTRY NUMBER
COMMENTARY
9075-08-5
not distinguished from EC 3.1.21.4
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
strain ATCC 10987, enzyme BceS1
SwissProt
Manually annotated by BRENDA team
EcoP15; EcoPI
-
-
Manually annotated by BRENDA team
EcoPI; enzyme EcoP15I
-
-
Manually annotated by BRENDA team
enzyme EcoK
-
-
Manually annotated by BRENDA team
enzyme EcoP1I
-
-
Manually annotated by BRENDA team
enzymes EcoP1I, EcoP15I
-
-
Manually annotated by BRENDA team
isoform EcoP15I
-
-
Manually annotated by BRENDA team
mutant enzymes: R.EcoPIK90R, R.EcoPIT91A and R.EcoPIH229K; R.EcoPI
-
-
Manually annotated by BRENDA team
Escherichia coli P15I
P15I
-
-
Manually annotated by BRENDA team
enzyme HinFIII
-
-
Manually annotated by BRENDA team
type III restriction-modification system
-
-
Manually annotated by BRENDA team
enzyme LlaFI
-
-
Manually annotated by BRENDA team
resNgoAXP restriction modification protein
UniProt
Manually annotated by BRENDA team
prophage P1
enzyme EcoP1I
-
-
Manually annotated by BRENDA team
; strain 164
-
-
Manually annotated by BRENDA team
Providencia stuartii 164
strain 164
-
-
Manually annotated by BRENDA team
strain GI-H
-
-
Manually annotated by BRENDA team
different strains
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
DNA cleavage by the type III restriction-modification enzymes requires communication in 1D between two distant indirectly-repeated recognitions sites, resulting in non-specific dsDNA cleavage close to only one of the two sites. The cleavage site selection reflects the dynamics of the preceding stochastic enzyme events that are consistent with bidirectional motion in 1D and DNA cleavage following head-on protein collision. The type III REs exhibits site orientation selectivity
physiological function
-
characterization of a type III-like restriction system present in clinical Staphylococcus aureus strains that prevents transformation with DNA from other bacterial species. Some methicillin-resistant strains are deficient in this restriction system, and thus are hypersusceptible to the horizontal transfer of DNA from other species, such as Escherichia coli, and could easily acquire a vancomycin-resistance gene from Enterococci, overview
additional information
-
sliding model for long-range communication on DNA by type III restriction enzymes, distribution of cleavage on head-to-head substrates with end bias, overview
additional information
-
several models for the mode of action of type III R/M enzymes, detailed overview. 1. Translocation, loop extrusion and collision model. 2. The end reversal model. 3. Transient looping and translocation model from atomic force spectroscopy
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
-
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
-
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
-
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
-
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
-
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
-
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
EcoP15 recognizes nonpalindromic DNA sequences, it requires a pair of nonmethylated inversely orientated recognition sites
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
the enzyme posseses helicase activity which may be involved in local unwinding of DNA in the cleavage sites
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
R.EcoPI has no strand separation activity
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
EcoP15 restriction is a very fast reaction that requires two unmodified recognition sites in inverted orientation. EcoP15 methylation is a slower reaction and is independent of the orientation of the sites
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
recognizes nonsymetric nucleotide sequences. A necessary condition for DNA cleavage is the presence of two unmethylated recognition sites which are inversely, head-to-head, oriented in the DNA double strand. A DNA substrate possessing one EcoP1 and one EcoP15 site in the head to head configuration can not be cleaved by an individual enzyme, however it is specifically digested in the simultaneously presence of both enzymes. The two different type III enzymes can functionally cooperate in the cleavage of DNA
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
restriction requires a pair of unmethylated, inversely oriented recognition sites. Suicidal restriction by EcoP15I is prevented if all the unmodified sites are in the same orientation. EcoP15I possesses an intrisic ATPase activity, the potential driving force of DNA translocation. The ATPase is recognition site-specific
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
action of type III restriction enzymes takes place on replicated or replicating DNA in VIVO and leaves daughter DNAs with breaks at nonallelic sites, that bacteriophage-mediated homologous recombinantion reconstitutes an intact DNA from them, and that REcBCD exonuclease blocks this repair by degradation from the restriction breaks
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
the intact Res2Mod2 tetramer is a fast endonuclease and slow methyltransferase, thereby favouring DNA cleavage, subassemblies of PstII in which the Res subunits have dissociated are more efficient methyltransferases. DNA cleavage by these lower molecular weight species may only occur if sufficient hsdR associates to form a Res2Mod2 tetramer before methylation occurs. This dynamic association of Res and Mod might play a key role in the control of PstII activity in vivo
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
cleavage site does not depend on the sequence context of the recognition sie. The enzyme can cleave linear DNA having either recognition sites in the same orientation or a single recognition site. Cleavage occurs predominantly at a site proximal to the DNA end in the case of multiple site substrates. The mechanism requires two enzyme molecules cooperating to elicit double-stranded break on DNA. The enzyme translocates on DNA in a 5' to 3' direction from its recognition site, switch in the direction of enzyme motion at the FNA ends
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
communication between type III recognition sites by energy-dependent 1D translocation and not by thermally driven 3D looping
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
ECoP15I recognizes the non-symmetric DNA sequence 5'-CAGCAG. For efficient cleavage, the enzyme needs the interaction with two copies of the recognition sequence that have to be inversely oriented in the DNA double-strand. The enzyme cuts the upper DNA strand 25-26 bp and the lower DNA strand 27-28 bp, respectively, downstream of the recognition sequence
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
recognition sequence: CTGATG. The enzyme requires a substrate with two copies of the recognition site in head-to-head repeat and is dependent on a low level of ATP hydrolysis (about 40 ATP/site/min). PstII can cut DNA with GTP and CTP
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
B0LX59
BsaHI recognises a palindromic sequence of bases and cleaves within this sequence between the purine and cytosine bases: GR/CGYC
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
EcoP15I rrequires the interaction with two inversely oriented 5'-CAGCAG recognition sites for efficient DNA cleavage
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
type III REs require two copies of their asymmetric recognition site in an indirectly repeated, head-to-head orientation cleaving the DNA 25-27 bp downstream of only one of the two sites
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
Geobacillus stearothermophilus CPW11
-
BsaHI recognises a palindromic sequence of bases and cleaves within this sequence between the purine and cytosine bases: GR/CGYC
-
-
?
phage lambda DNA + H2O
?
show the reaction diagram
-, Q5F957
restriction activity requires the presence of both the restriction modification endonuclease protein and the methylase protein of the NgoAXP restriction modification system
-
-
-
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
Providencia stuartii 164
-
-, the intact Res2Mod2 tetramer is a fast endonuclease and slow methyltransferase, thereby favouring DNA cleavage, subassemblies of PstII in which the Res subunits have dissociated are more efficient methyltransferases. DNA cleavage by these lower molecular weight species may only occur if sufficient hsdR associates to form a Res2Mod2 tetramer before methylation occurs. This dynamic association of Res and Mod might play a key role in the control of PstII activity in vivo, recognition sequence: CTGATG. The enzyme requires a substrate with two copies of the recognition site in head-to-head repeat and is dependent on a low level of ATP hydrolysis (about 40 ATP/site/min). PstII can cut DNA with GTP and CTP
-
-
?
additional information
?
-
-
the database REBASE contains information about recognition sites and cleavage sites
-
-
-
additional information
?
-
-
cleavage also occurs for two adjacent head to head and tail to tail oriented target sites, but only at one of the two possible sites, DNA translocation is not required
-
?
additional information
?
-
prophage P1
-
cleavage generally depends on ratio of enzyme to cleavage sites, in presence of S-adenosyl-L-methionine, substrates with two cleavage sites in inverted repeats are susceptible, in absence of S-adenosyl-L-methionine, presence of Na+ leads to cleavage only in substrates with two sites in inverted repeats, presence of K+ leads to cleavage of all substrates
-
?
additional information
?
-
-
cooperation of enzyme with EcoP15I in DNa cleavage, cleavage predominantly at EcoP15I site, EcpP1I greatly stimulates EcoP15I nicking activity
-
?
additional information
?
-
prophage P1
-
cooperation of enzyme with EcoP1I in DNa cleavage, cleavage predominantly at EcoP15I site, EcoP1I greatly stimulates EcoP15I nicking activity
-
?
additional information
?
-
-
structural requirements of substrate, overview
-
?
additional information
?
-
-
the enzyme uses both diffuse DNA loop formation and ATPase driven translocation of the intervening DNA contour to communicate between two recognition sites
-
-
-
additional information
?
-
-
type III restriction-modification systems play only a minor role in the overall defence of the cell against invasion by foreign DNA
-
-
-
additional information
?
-
-
the pre-incubation of EcoP15I with DNA has no significant influence on the enzyme's ability to cleave DNA
-
-
-
additional information
?
-
-
on linear DNA in contrast, there is only one route and resolvase completely blocks communication and cleavage, while on circular DNA, there are two routes for communication. If one route is blocked by resolvase, the enzyme can use the alternative route. No stepwise motor mechanism for type III enzymes, the enzyme can bypass triplexes during sliding, DNA end capping stimulates cleavage of tail-to-tail oriented pairs of sites, rates of cleavage of tail-to-tail repeats, ATP hydrolysis rates, and DNA cleavage kinetics, overview
-
-
-
additional information
?
-
-
EcoP15I recognizes 5'-CAGCAG-3' as target site. Cleavage occurs at a defined location next to only one of the sites, and one of the two sites is chosen for cleavage randomly, although this is influenced by the base composition of the DNA. The EcoP15I R/M enzyme cuts the sequence CAGCAG(N)25/27 as long as the underlined adenine is unmethylated. Preference for a two-site substrate. ATP hydrolysis is essential for the overall restriction process
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
-
action of type III restriction enzymes takes place on replicated or replicating DNA in VIVO and leaves daughter DNAs with breaks at nonallelic sites, that bacteriophage-mediated homologous recombinantion reconstitutes an intact DNA from them, and that REcBCD exonuclease blocks this repair by degradation from the restriction breaks
-
-
?
DNA + H2O
specific double-stranded DNA fragments with terminal 5'-phosphate
show the reaction diagram
Providencia stuartii, Providencia stuartii 164
-
the intact Res2Mod2 tetramer is a fast endonuclease and slow methyltransferase, thereby favouring DNA cleavage, subassemblies of PstII in which the Res subunits have dissociated are more efficient methyltransferases. DNA cleavage by these lower molecular weight species may only occur if sufficient hsdR associates to form a Res2Mod2 tetramer before methylation occurs. This dynamic association of Res and Mod might play a key role in the control of PstII activity in vivo
-
-
?
additional information
?
-
-
the enzyme uses both diffuse DNA loop formation and ATPase driven translocation of the intervening DNA contour to communicate between two recognition sites
-
-
-
additional information
?
-
-
type III restriction-modification systems play only a minor role in the overall defence of the cell against invasion by foreign DNA
-
-
-
additional information
?
-
-
on linear DNA in contrast, there is only one route and resolvase completely blocks communication and cleavage, while on circular DNA, there are two routes for communication. If one route is blocked by resolvase, the enzyme can use the alternative route. No stepwise motor mechanism for type III enzymes, the enzyme can bypass triplexes during sliding
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ATP
-
ATP-dependent type III restriction endonuclease
S-adenosyl-L-methionine
-
endogenously bound, absolutely required
S-adenosyl-L-methionine
prophage P1
-
allosteric activator and specificity factor that ensures that cleavage only occurs when two endonucleases bind two recognition sites in a designated orientation
S-adenosyl-L-methionine
-
S-adenosyl-L-methionine is not necessary for DNA cleavage, the presence of S-adenosyl-L-methionine dramatically impaires DNA cleavage due to competing DNA methylation
S-adenosyl-L-methionine
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mg2+
-
required
Mg2+
P25241
required
Mg2+
-
required for activity
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
HU protein
-
binding of HU protein interfers with R.EcoP15I cleavage activity
-
Lac repressor
-
cleavage can be abolished by the binding of Lac repressor downstream (3' side) but not upstream (5' side) of the recognition site
-
additional information
-
type III restriction is alleviated by bacteriophage (RecE) homologous recombination function
-
additional information
-
sinefungin neither has an appreciable effect on DNA cleavage by EcoP15I nor compensated for the second recognition site; the catalytic activity of EcoP15I after 3 h of pre-incubation without DNA is strongly decreased
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
N4-adenosyl-N4-methyl-2,4-diaminobutanoic acid
-
may substitute for S-adenosyl-L-methionine
S-adenosylmethionine
-
stimulates
S6-methyl-6-deaminosinefungin
-
may substitute for S-adenosyl-L-methionine
Sinefungin
-
may substitute for S-adenosyl-L-methionine
additional information
-
type III restriction is enhanced by bacterial (RecBCD) function
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8
-
-
assay at
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
assay at
80
-
-
the optimal temperature for DNA cleavage by PspGI is at 80°C or above
PDB
SCOP
CATH
ORGANISM
Bacillus subtilis (strain 168)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
77500
-
-, Q5F957
calculated
407000
-
-
sedimentation studies
409000
-
prophage P1
-
sedimentation studies
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
tetramer
Escherichia coli, prophage P1
-
2 * 105000 + 2 * 74000, deduced from gene sequence
tetramer
-
the intact Res2Mod2 tetramer is a fast endonuclease and slow methyltransferase, thereby favouring DNA cleavage, subassemblies of PstII in which the Res subunits have dissociated are more efficient methyltransferases. DNA cleavage by these lower molecular weight species may only occur if sufficient hsdR associates to form a Res2Mod2 tetramer before methylation occurs. This dynamic association of Res and Mod might play a key role in the control of PstII activity in vivo
tetramer
Providencia stuartii 164
-
the intact Res2Mod2 tetramer is a fast endonuclease and slow methyltransferase, thereby favouring DNA cleavage, subassemblies of PstII in which the Res subunits have dissociated are more efficient methyltransferases. DNA cleavage by these lower molecular weight species may only occur if sufficient hsdR associates to form a Res2Mod2 tetramer before methylation occurs. This dynamic association of Res and Mod might play a key role in the control of PstII activity in vivo
-
?
-
x * 106000, subunit Res, + x * 75000, subunit Mod, deduced from gene sequence
additional information
-
separation of Res and Mod subunits by gel filtration in presence of 2 M NaCl
additional information
-
functional interaction of enzymes EcoP1I and EcoP15I
additional information
-
in the presence of specific DNA, the entire modification subunit of the enzyme is protected from trypsin digestion, whereas in the absence of DNA stable protein domains of the modification subunit are not detected. In contrast, the restriction subunit is comprised of two trypsin-resistant domains of about 77000-79000 Da and 27000-29000 Da, respectively. Both structural restriction domains are connected by a flexible linker region that spans 23 amino acid residues
additional information
-
the type III R/M enzymes comprise two modification subunits, each containing a target recognition domain to bind to the target sequence and a methyltransferase catalytic domain to monitor the methylation status of an adenine in the target, and two restriction subunits each containing a DNA helicase and ATP-hydrolysing domain and an endonuclease DNA cleavage domain
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
database information: http://rebase.neb.com
-
real-time imaging of enzyme at scan rates of 1-3 frames per s. EcoP15I translocates DNA in an ATP-dependent manner, at a rate of 79 bp/s, resulting in the accumulation of supercoiling. EcoP15I bound to its recognition site also makes nonspecific contacts with other DNA sites, thus forming DNA loops and reducing the distance between the two recognition sites
-
study of enzyme-DNA pre-cleavage complexes by atomic force microscopy. DNA loops observed are formed by a contact between site-bound EcoP15I enzyme and a nonspecific region of DNA, and do not result from translocation. Model for restricition by type III enzymes involving both structural elements, and a functional reason for the unusual site orientation required for the cleavage reaction
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
95
-
-
half life of 2 h at 95°C
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
EcoP15I is stabilized in the presence of specific DNA
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Ni-NTA agarose column chromatography
B0LX59
P11 cellulose phosphate column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
cloned into pQE-16 plasmid vector that provides the enzyme with a C-terminal 6His-tag
-
expression in Escherichia coli
-
expressed in Escherichia coli strain ER2566
B0LX59
expression in Escherichia coli
-, Q5F957
expressed in Escherichia coli strain ER2744
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D898A
-
Res subunit, no enzymic activity
E916A
-
Res subunit, no enzymic activity
G448S
-
mutant of Mod subunit, not able to bind S-adenosyl-L-methionine, no DNA cleavage
R534A
-
Res subunit, no enzymic activity
D354A
B0LX59
mutant shows reduced activity compared to the wild type enzyme
E350A
B0LX59
mutant shows reduced activity compared to the wild type enzyme
F353A
B0LX59
mutant shows reduced activity compared to the wild type enzyme
P349A
B0LX59
mutant shows reduced activity compared to the wild type enzyme
Q344A
B0LX59
mutant shows activity similar to wild type enzyme
R351A
B0LX59
the activity of the R351A mutant is negligible
R352A
B0LX59
mutant shows activity similar to the wild type enzyme
D898A
prophage P1
-
Res subunit, no enzymic activity
E916A
prophage P1
-
Res subunit, no enzymic activity
K918A
prophage P1
-
Res subunit, no enzymic activity
P897A
prophage P1
-
Res subunit, activity similar to wild type
R534A
prophage P1
-
Res subunit, no enzymic activity
D138A
-
catalytically inactive
K918A
-
Res subunit, no enzymic activity
additional information
-
mutant c2-134, unable to cleave DNA, mutant c2-440, poor ability to cleave DNA
additional information
-
study on the predicted linker region between the two domains of the restriction subunit by insertional mutagenesis. Introduction of up to 18 amino acids in the N- and C-terminal region of the linker. The region tolerated the introduced genetic alterations without loss of catalytic function or changes in cleavage position
S348A
B0LX59
mutant shows activity similar to the wild type enzyme
additional information
-
analysis of several strains for phase variable methyltransferase activity, mod, and restriction endonuclease activity. Of 41 strains in the survey that contain a phase variable mod gene, seven have an obvious mutation in the restriction domain and appear to be dedicated phasevarions. The remaining 15 strains that do not contain a phase variable mod gene are likely to be dedicated, functional, type III restriction-modification systems
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
reconstitution of separated Res and Mod subunits leads to an inactive apoenzyme lacking S-adenosyl-L-methionine, reconstitution in presence of S-adenosyl L-methionine was not successful
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
analysis
-
transcriptome analysis
additional information
-
inactivation of this restriction system dramatically increases the transformation efficiency of clinical Staphylococcus aureus methicillin-resistant strains, opening the field of molecular genetic manipulation of these strains using DNA of exogenous origin