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Information on EC 2.1.1.72 - site-specific DNA-methyltransferase (adenine-specific) and Organism(s) Escherichia coli and UniProt Accession P0AEE8
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2.1.1.72 site-specific DNA-methyltransferase (adenine-specific)Specify your search results
Word Map
- 2.1.1.72
- mtases
- n6-methyladenosine
- endonuclease
- chromatin
- duplex
- s-adenosyl-l-methionine
- cytosine
- adomet
- bacteriophage
- mismatch
-
damid
-
unmethylated
-
hemimethylated
-
restriction-modification
- flip
-
protein-dna
-
methylation-sensitive
- s-adenosyl-l-homocysteine
- 2-aminopurine
-
methyltransferase-like
-
epitranscriptomic
-
writer
- m.hhai
-
merip-seq
-
mettl3-mediated
- alkbh5
- crescentus
- medicine
- prophage
-
m6a-dependent
-
adohcy
-
m6a-modified
-
gatatc
-
n6-adenosine
- sinefungin
-
n6-methylation
-
extrahelical
-
methyl-directed
- aquaticus
- analysis
- pharmacology
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
+
adenine in DNA
=
+
N6-methyladenine in DNA
Synonyms
dam mtase, dna adenine methyltransferase, m6a methyltransferase, dam methylase, dna adenine methylase, m.taqi, t4 dam, specific methyltransferase, m.ecorv, ecodam, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
adenine-N6 MTAse
-
-
-
-
Dam
DNA adenine methyltransferase
DNA-[N6-adenine]-methyltransferase
-
-
-
-
modification methylase
-
-
-
-
N6-adenine DNA -methyltransferase
-
-
-
-
restriction-modification system
-
-
-
-
Dam
-
-
DNA adenine methyltransferase
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + adenine in DNA = S-adenosyl-L-homocysteine + N6-methyladenine in DNA
S-adenosyl-L-methionine + adenine in DNA = S-adenosyl-L-homocysteine + N6-methyladenine in DNA
D181 and Y184 are essential for activity
-
S-adenosyl-L-methionine + adenine in DNA = S-adenosyl-L-homocysteine + N6-methyladenine in DNA
ordered bi bi mechanism with S-adenosyl-L-methionine binding first, fast methyl transfer, slow product release
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
methyl group transfer
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
69553-52-2
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + adenine in DNA
S-adenosyl-L-homocysteine + DNA containing N6-methyladenine
-
the enzyme specifically binds double-stranded DNA
-
-
?
S-adenosyl-L-methionine + adenine in DNA
S-adenosyl-L-homocysteine + N6-methyladenine in DNA
-
-
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
S-adenosyl-L-methionine + DNA duplex carrying the 5'-AAGCUU-3'/3'-TTCGAA-5' target sequence
?
-
100% activity
-
-
-
S-adenosyl-L-methionine + double stranded DNA adenine
S-adenosyl-L-homocysteine + double stranded DNA 6-methyladenine
-
-
-
-
?
S-adenosyl-L-methionine + pUC19 DNA adenine
S-adenosyl-L-homocysteine + pUC19 DNA 6-methyladenine
-
-
-
-
?
S-adenosyl-L-methionine + RNA/DNA heteroduplex carrying the 5'-AAGCUU-3'/3'-TTCGAA-5' target sequence
?
-
low activity. Only the DNA strand of the RNA/DNA heteroduplex is methylated
-
-
-
S-adenosyl-L-methionine + single stranded DNA adenine
S-adenosyl-L-homocysteine + single stranded DNA 6-methyladenine
-
the level of single stranded DNA methylation is 7fold lower than double stranded DNA. However, upon star activity conditions (30% DMSO), the efficiency of single stranded DNA modification is raised by up to 50% of the relative activity
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
Dam methylates the N-6 position of the adenine in the sequence 5'-GATC-3', Dam shows a dramatic preference for the in vitro methylation of certain GATC sequences in plasmids and PCR-derived DNAfragments
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
-
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
-
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
Dam belongs to the alpha-class of adenine methyltransferases and transfers a methyl group to the N-6 position of the adenine in the DNA sequence 5'-GATC-3'
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
Dam methylates a GATC recognition site
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
Dam methylates the adenine residue in GATC sites
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
Dam methylates the N-6 position of adenine in the DNA sequence 5'-GATC-3' and is highly processive, catalyzing multiple methyltransfers prior to dissociating from the DNA
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
DNA adenine methyltransferase methylates the N6 positions of adenines in the sequence 5'-GATC-3'
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
EcoP15I MTase adds a methyl group to the second adenine in the recognition sequence 5'-CAGCAG-3' in the presence of S-adenosyl-L-methionine
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
the DNA target sequence is GATC, the natural substrate for the enzyme is hemimethylated DNA, where one strand is methylated and the other is not
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
the target sequence is 5'GATC3'
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
the wild type enzyme shows target specificity for GATC sites
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
the EcoRV DNA methyltransferase methylates the first adenine in the GATATC recognition sequence
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
the enzyme methylates adenines at the N6 position of palindromic 5-GATC-3 sites. The enzyme ethylates both strands of the same site prior to fully dissociating from the DNA, a process referred to as intrasite processivity. Intrasite processivity is disrupted when the DNA flanking a single GATC site is longer than 400 bp on either side. The introduction of a second GATC site within this flanking DNA reinstates intrasite methylation of both sites
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
the enzyme recognizes the palindromic specific sequence 5'-AAGCTT-3' and catalyzes formation of N6-methyladenine at the first A-residue. The enzyme prefers DNA containing a hemimethylated target site
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
-
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
canonical 14mers and various substituted duplexes. Non-selfcomplementary tetradecamer duplex d[GCCGGATCTAGACG]*d[CGTCTAGATCCGGC] containing the hemimethylated GATC target sequence on one or the other strand and modifications in the GATC target sequence of the complementary strands. Large differences in DNA methylation of duplexes containing single dI or dG substitutions of the Dam recognition site are observed compared with the canonical substrate, if the substitution involves the top strand, on the G-C rich side. Substitution in either strand by uracil or 5-ethyluracil result in small perturbation of the methylation patterns. When 2,6-diamino-purine replaces the adenine to be methylated, small but significant methylation is observed
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
non-self-complementary tetradecanucleotide duplexes that contain the GATC target sequence. The enzyme is rather tolerant to base modification, binding of the enzyme is inversely proportional to the thermodynamic stability of the duplex in the ternary complex
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
methylation of DNA in a sequence specific manner, low substrate specificity with respect to the target base. Cytosine residues can be methylated if they are located in a C:T mismatch base pair at the target position of the enzyme, modification of cytosine residues at position N4
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
enzyme is a critical regulator of bacterial virulence
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
methylation of the site GATCprox proximal to the promotor is required for transition to the phase On state by specifically blocking PapI-dependent binding of Lrp to promotor proximal sites 4-6, expression of pyelonephritis-associated pili, i.e. Pap, in uropathogenic Escherichia coli is epigenetically controlled by a reversible OFF to ON switch, overview
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
the enzyme is cell cycle regulated and essential for viability
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
DNA substrate from calf thymus, methylation of adjacent GATC sites is distributive with DNA derived from a genetic element that controls the transcription of the adjacent genes, the first methylation event is followed by enzyme release
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
enzyme methylates the first adenine in the sequence ATGCAT
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
methylation of target sequence GATC
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
required substrate recognition target sequence is GATC, occuring base flipping in absence of S-adenosyl-L-methionine is a biphasic process and very fast, but binding of the flipped base into the active site pocket requiring S-adenosyl-L-methionine is slow, active site contains the conserved DPPY motif, whose tyrosine184 residue stacks to the flipped target base
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
specific for methylation site sequence GATC and structure, an intact GA sequence is essential for activity, altered structural symmetry of the DNA substrate decreases kcat sharply, the best contact between enzyme and DNA is a palindromic interaction site covering the 5'-symmetric residues, which is located in the major groove, and another one in the 3'-half covering the 3'-symmetric residues is located in the minor groove, overview
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
transition of enzyme-DNA interaction from nonspecific to specific interaction utilizing different substrates, identification of discriminatory contacts stabilizing the transition state, and antidiscriminatory contacts not affecting the methylation of the cognate site but disfavor activity at noncognate sites, involved are Y119, N120, L122, R124, and P134, overview
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
the enzyme recognizes 5'-GATC-3'
-
-
?
additional information
?
-
the wild type enzyme is active on hemimethylated and unmethylated DNA substrates
-
-
?
additional information
?
-
relative to GATC, three near-cognate substrates that carry a base-pair substitution at the first position are still methylated by the enzyme, although at rates reduced by 100fold (AATC) or 1000fold (CATC)
-
-
?
additional information
?
-
-
relative to GATC, three near-cognate substrates that carry a base-pair substitution at the first position are still methylated by the enzyme, although at rates reduced by 100fold (AATC) or 1000fold (CATC)
-
-
?
additional information
?
-
-
substrate specificity with different oligonucleotides, substrate binding specificity, overview
-
-
?
additional information
?
-
-
catalyzes the transfer of a methyl group to the C5 position of the 3'-side cytosine of each strand of the recognition sequence, M.EcoRII binding is diminished by factors of 5-30 but the catalytic activity is either abolished or reduced 4-80fold when trans-anti-B[a]P-N2-dG lesions are introduced into the EcoRII recognition sequence, methylation rates are also diminished and in some cases entirely abolished, depending on the position of the lesion within the recognition sequence
-
-
?
additional information
?
-
-
DNA methylation by DAM may not globally affect gene transcription by physically blocking access of transcription factors to binding sites, Dam is down regulated in the stationary phase, which correlates with the enrichment of GATC in binding sites for CRP and Sigma 38
-
-
?
additional information
?
-
-
the preferred substrate consists of the annealed product of oligonucleotides 5'-CATTTACTTGATCCGGTATGC-3' and 5'-GCATACCGGATCAAGTAAATG-3', while the nonpreferred substrate consists of the annealed product of oligonucleotides 5'-CATTTAGACGATCTTTTATGC-3' and 5'-GCATAAAAGATCGTCTAAATG-3'
-
-
?
additional information
?
-
-
the wild type Dam shows no detectable activity at GATT sites
-
-
?
additional information
?
-
-
the DNA substrate preference for M.EcoVIII is: hemimethylated AAGCTT > AAGCTT > GAGCTT >/= CAGCTT > AATCTT > ATGCTT > ACGCTT > TAGCTT > AACCTT = single stranded AAGCTT > AGGCTT = AAACTT
-
-
-
additional information
?
-
-
the enzyme is not able to methylate nucleotide sequence containing two mismatches (5'-CAGCTG-3')
-
-
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
Dam methylates the N-6 position of the adenine in the sequence 5'-GATC-3', Dam shows a dramatic preference for the in vitro methylation of certain GATC sequences in plasmids and PCR-derived DNAfragments
-
-
?
S-adenosyl-L-methionine + adenine in DNA
S-adenosyl-L-homocysteine + DNA containing N6-methyladenine
-
the enzyme specifically binds double-stranded DNA
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
-
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
-
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
Dam belongs to the alpha-class of adenine methyltransferases and transfers a methyl group to the N-6 position of the adenine in the DNA sequence 5'-GATC-3'
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
Dam methylates a GATC recognition site
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
Dam methylates the adenine residue in GATC sites
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
Dam methylates the N-6 position of adenine in the DNA sequence 5'-GATC-3' and is highly processive, catalyzing multiple methyltransfers prior to dissociating from the DNA
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
DNA adenine methyltransferase methylates the N6 positions of adenines in the sequence 5'-GATC-3'
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
EcoP15I MTase adds a methyl group to the second adenine in the recognition sequence 5'-CAGCAG-3' in the presence of S-adenosyl-L-methionine
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
the DNA target sequence is GATC, the natural substrate for the enzyme is hemimethylated DNA, where one strand is methylated and the other is not
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
the target sequence is 5'GATC3'
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
the wild type enzyme shows target specificity for GATC sites
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
the enzyme recognizes the palindromic specific sequence 5'-AAGCTT-3' and catalyzes formation of N6-methyladenine at the first A-residue. The enzyme prefers DNA containing a hemimethylated target site
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
-
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
enzyme is a critical regulator of bacterial virulence
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
methylation of the site GATCprox proximal to the promotor is required for transition to the phase On state by specifically blocking PapI-dependent binding of Lrp to promotor proximal sites 4-6, expression of pyelonephritis-associated pili, i.e. Pap, in uropathogenic Escherichia coli is epigenetically controlled by a reversible OFF to ON switch, overview
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
the enzyme is cell cycle regulated and essential for viability
-
-
?
S-adenosyl-L-methionine + DNA adenine
S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
the enzyme recognizes 5'-GATC-3'
-
-
?
additional information
?
-
the wild type enzyme is active on hemimethylated and unmethylated DNA substrates
-
-
?
additional information
?
-
-
the preferred substrate consists of the annealed product of oligonucleotides 5'-CATTTACTTGATCCGGTATGC-3' and 5'-GCATACCGGATCAAGTAAATG-3', while the nonpreferred substrate consists of the annealed product of oligonucleotides 5'-CATTTAGACGATCTTTTATGC-3' and 5'-GCATAAAAGATCGTCTAAATG-3'
-
-
?
additional information
?
-
-
the wild type Dam shows no detectable activity at GATT sites
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
diethyl dicarbonate
-
the inactivation of the enzyme by diethyl dicarbonate is specific for histidine residues, pre-incubating the methylase with DNA is able to protect the enzyme from diethyl dicarbonate inactivation, hydroxylamine is unable to reverse the effect caused by diethyl dicarbonate
methylated DNA
-
mixed-type inhibition against S-adenosyl-L-methionine, uncompetitive against unmethylated DNA
-
S-adenosyl-L-homocysteine
-
competitive against S-adenosyl-L-methionine, uncompetitive against DNA
additional information
-
5'-allylthio-5'-deoxy-9-(1'-beta-D-ribofuranosyl)6-nitrodideazaadenine, 5'-chloro-5'-deoxyadenosine, 9-(1'-beta-D-ribofuranosyl)6-nitro-1,3-dideazaadenine, 5'-S-(propionamide)5'-deoxy-9-(1'-beta-D-ribofuranosyl)-adenine, 5'-S-(ethylpropionate)5'-deoxy-9-(1'-beta-D-ribofuranosyl)adenine, 5'-S-5'-methylthio deoxyadenosine, 5'-S-propargylthio adenosine, 5'-S-allenylthio adenosine, and 5'-S-propynylthioadenosine have no effect on M.EcoP15I activity
-
additional information
-
imidazole (5 mM) and ethanol (0.02% v/v) do not show any effect on enzyme
-
additional information
-
leucine-responsive regulatory protein specifically inhibits Dam methylation of GATC proximal to the pilus genes when pre-assembled onto a pap half-site substrate and when competing kinetically with Dam for access to this site, leucine-responsive regulatory protein does not efficiently inhibit Dam methylation of non-regulatory GATC sites in plasmid DNA, leucine-responsive regulatory protein inhibits Dam methylation of GATC proximal to the pilus genes in a pap substrate where GATC distal to the pilus genes is hemimethylated
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
5'-S-(propionic acid)5'-deoxy-9-(1'-beta-D-ribofuranosyl)1,3-dideazaadenine
-
more than 200% activity in the presence of 0.015 mM 5'-S-(propionic acid)5'-deoxy-9-(1'-beta-D-ribofuranosyl)1,3-dideazaadenine
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0002576
single stranded DNA adenine
-
with specific sequence AAGCTT, at pH 7.0 and 37°C
-
0.0000227
DNA adenine
using the DNA sequence 5'-GCATACCCGATCAAGTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.0000809
DNA adenine
using the DNA sequence 5'-GCATAAAAGATCGTCTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.000007
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R116A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.000008
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R95A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.00001
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme N132A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.000023
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.000027
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme K139A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.000031
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme N126A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.000035
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R116A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.000044
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme K139A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.000076
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme N132A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.000081
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.000083
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R137A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.000086
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R95A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.00022
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R137A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.0003
DNA adenine
-
in 50 mM NaCl, 50 mM HEPES, 1 mM EDTA, pH 7.5, temperature not specified in the publication
0.0000244
double stranded DNA adenine
-
with specific sequence MAGCTT/TTCGAA, at pH 7.0 and 37°C
-
0.0000751
double stranded DNA adenine
-
with specific sequence AAGCTT/TTCGAA, at pH 7.0 and 37°C
-
0.0000782
double stranded DNA adenine
-
with specific sequence CAGCTT/GTCGAA, at pH 7.0 and 37°C
-
0.0000784
double stranded DNA adenine
-
with specific sequence GAGCTT/CTCGAA, at pH 7.0 and 37°C
-
0.0000855
double stranded DNA adenine
-
with specific sequence TAGCTT/ATGCAA, at pH 7.0 and 37°C
-
0.000089
double stranded DNA adenine
-
with specific sequence ACGCTT/TGCGAA, at pH 7.0 and 37°C
-
0.0000912
double stranded DNA adenine
-
with specific sequence AGGCTT/TCCGAA, at pH 7.0 and 37°C
-
0.0000914
double stranded DNA adenine
-
with specific sequence ATGCTT/TACGAA, at pH 7.0 and 37°C
-
0.0001023
double stranded DNA adenine
-
with specific sequence AAACTT/TTTGAA, at pH 7.0 and 37°C
-
0.0001273
double stranded DNA adenine
-
with specific sequence AACCTT/TTGGAA, at pH 7.0 and 37°C
-
0.000143
double stranded DNA adenine
-
with specific sequence AATCTT/TTAGAA, at pH 7.0 and 37°C
-
0.012
S-adenosyl-L-methionine
-
in 50 mM NaCl, 50 mM HEPES, 1 mM EDTA, pH 7.5, temperature not specified in the publication
additional information
additional information
-
Km-values fpr 14-mers and various substituted duplexes
-
additional information
additional information
-
turnover numbers for the canonical 14-mer duplex and various substituted duplexes
-
additional information
additional information
-
detailed kinetics for diverse substrate duplexes and methylation sites
-
additional information
additional information
-
quantitative stopped-flow kinetics using 2-aminopurine as a probe to detect base flipping, wild-type and mutant enzymes
-
additional information
additional information
-
steady-state kinetics, kinetic mechanism, thermodynamics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0005
single stranded DNA adenine
-
with specific sequence AAGCTT, at pH 7.0 and 37°C
-
0.0038
DNA adenine
using the DNA sequence 5'-GCATACCCGATCAAGTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.015
DNA adenine
using the DNA sequence 5'-GCATAAAAGATCGTCTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.00017
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme N132A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.00082
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R95A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.0009
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R116A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.0015
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme N126A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.003
DNA adenine
-
in 100 mM Tris (pH 8.0), 1 mM EDTA, 1 mM dithiothreitol, and 0.2 mg/ml bovine serum albumin, at 22°C
0.0038
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.0055
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme K139A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.008
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme K139A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.0083
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R95A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.013
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R116A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.015
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.019
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R137A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.02
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme N132A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.058
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R137A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
0.0002
double stranded DNA adenine
-
with specific sequence AGGCTT/TCCGAA, at pH 7.0 and 37°C
-
0.00022
double stranded DNA adenine
-
with specific sequence ACGCTT/TGCGAA, at pH 7.0 and 37°C
-
0.00022
double stranded DNA adenine
-
with specific sequence TAGCTT/ATGCAA, at pH 7.0 and 37°C
-
0.00025
double stranded DNA adenine
-
with specific sequence ATGCTT/TACGAA, at pH 7.0 and 37°C
-
0.00027
double stranded DNA adenine
-
with specific sequence AACCTT/TTGGAA, at pH 7.0 and 37°C
-
0.00033
double stranded DNA adenine
-
with specific sequence AAACTT/TTTGAA, at pH 7.0 and 37°C
-
0.00033
double stranded DNA adenine
-
with specific sequence AATCTT/TTAGAA, at pH 7.0 and 37°C
-
0.00033
double stranded DNA adenine
-
with specific sequence CAGCTT/GTCGAA, at pH 7.0 and 37°C
-
0.00033
double stranded DNA adenine
-
with specific sequence GAGCTT/CTCGAA, at pH 7.0 and 37°C
-
0.0015
double stranded DNA adenine
-
with specific sequence AAGCTT/TTCGAA, at pH 7.0 and 37°C
-
0.0021
double stranded DNA adenine
-
with specific sequence MAGCTT/TTCGAA, at pH 7.0 and 37°C
-
additional information
additional information
-
turnover numbers for the canonical 14-mer duplex and various substituted duplexes
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.9
single stranded DNA adenine
-
with specific sequence AAGCTT, at pH 7.0 and 37°C
-
168
DNA adenine
using the DNA sequence 5'-GCATACCCGATCAAGTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
187
DNA adenine
using the DNA sequence 5'-GCATAAAAGATCGTCTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
48.3
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme N126A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
100
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R95A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
117
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R95A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
125
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme K139A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
133
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R116A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
160
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme N132A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
167
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
187
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
223
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R137A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
262
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R137A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
267
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme N132A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
300
DNA adenine
using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme K139A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
367
DNA adenine
using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R116A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C
1.7
double stranded DNA adenine
-
with specific sequence AAACTT/TTTGAA, at pH 7.0 and 37°C
-
1.7
double stranded DNA adenine
-
with specific sequence AGGCTT/TCCGAA, at pH 7.0 and 37°C
-
1.9
double stranded DNA adenine
-
with specific sequence AACCTT/TTGGAA, at pH 7.0 and 37°C
-
2.2
double stranded DNA adenine
-
with specific sequence TAGCTT/ATGCAA, at pH 7.0 and 37°C
-
2.6
double stranded DNA adenine
-
with specific sequence ACGCTT/TGCGAA, at pH 7.0 and 37°C
-
2.7
double stranded DNA adenine
-
with specific sequence ATGCTT/TACGAA, at pH 7.0 and 37°C
-
2.9
double stranded DNA adenine
-
with specific sequence AATCTT/TTAGAA, at pH 7.0 and 37°C
-
4.2
double stranded DNA adenine
-
with specific sequence CAGCTT/GTCGAA, at pH 7.0 and 37°C
-
4.3
double stranded DNA adenine
-
with specific sequence GAGCTT/CTCGAA, at pH 7.0 and 37°C
-
19.6
double stranded DNA adenine
-
with specific sequence AAGCTT/TTCGAA, at pH 7.0 and 37°C
-
86.1
double stranded DNA adenine
-
with specific sequence MAGCTT/TTCGAA, at pH 7.0 and 37°C
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
the enzyme, in addition to being responsible for GATC methylation, can also function as a methylation-independent transcriptional repressor
physiological function
physiological function
-
Dam is a key regulator of the pap operon which codes for the pilus proteins necessary for uropathogenic Escherichia coli cellular adhesion. Lrp, in the presence and in the absence of PapI and nonspecific DNA, specifically protects pap regulatory GATC sites from Dam methylation when allowed to compete with Dam for assembly on unmethylated and hemimethylated pap DNA. Only at low Lrp concentrations will Dam compete effectively for binding and methylation of the proximal GATC site, leading to a phase switch resulting in the expression of pili
physiological function
-
DNA adenine methyltransferase is essential for proper DNA replication timing, gene regulation, and mismatch repair
physiological function
-
synthesis of P pili (pyelonephritis-associated protein) is subjected to phase variation and switching between On and Off states is controlled by Dam methylation, Dam methylation inhibits OxyR binding in the On state, Dam methylation of the upstream GATC site in agn43 increases transcription initiation, which occurs precisely at the G nucleotide of the GATC, finP transcription is activated by Dam methylation, Dam methylation does not regulate tir gene transcription nor tir mRNA stability
physiological function
-
the formation of 6-methyladenine reduces the thermodynamic stability of DNA and changes DNA curvature, as a consequence, the methylation state of specific adenosine moieties can affect DNA-protein interactions. Dam methylation regulates virulence genes at the posttranscriptional level. Dam methylation plays a role in enterohemorrhagic Escherichia coli by controlling the production of the virulence factor Shiga toxin 2
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
additional information
dimer
-
Dam dimerizes on short, synthetic DNA, resulting in enhanced catalysis, however, dimerization is not observed on large genomic DNA
additional information
-
structure analysis of enzyme in ternary complex with partially and fully specific DNA and a methyl-donor analogue
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
in complex with cognate DNA at 1.89 A resolution, in the presence of S-adenosyl-L-homocysteine
in complex with non-cognate DNA, lacking any GATC sequences, hanging drop vapor diffusion method, using 5-40% (w/v) PEG 200 from and 100 mM MES or HEPES (pH 6.4-7.0)
in complex with duplex DNA and AMP, hanging drop vapor diffusion method, using 10% (w/v) PEG 5000 monomethyl ether, 0.1 M HEPES pH 7.5, 0.2M potassium acetate and 15 mM MnCl2 at 20°C
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
L122A
L122A/P134A
the variant methylates hemimethylated DNA with 80% of wild type activity
L122A/V133L
L122G
the mutant has no preference for hemimethylated substrate and is about 1.5fold more active than the wild type enzyme on the hemimethylated DNA substrate
L122I
the variant methylates hemimethylated DNA with 98% of wild type activity
L122S
the variant is able to sense the methylation status of the 5'-GATC-3' double-stranded target recognition site and methylates only hemimethylated DNA with 35-40% of wild type activity
L122T
the variant methylates hemimethylated DNA with 90% of wild type activity
L122V
the variant methylates hemimethylated DNA with 95% of wild type activity
N120A
loses its p-stacking with Gua1, shows small changes in specificity factor S1
P134A
P134G
high catalytic activity, exhibits only a small reduction in the amplitude of the fluorescence change, but no detectable changes in the kinetics of base-flipping, induces base-flipping of the substrate with altered sequence at the third base-pair
R124A
overall reduction in catalytic activity but methylates the near-cognate substrates GATT and GATG faster than the canonical GATC, no base-flipping signal
V133A
the mutant shows increased activity with hemimethylated DNA compared to the wild type enzyme (160%)
V133I
the mutant shows decreased activity with hemimethylated DNA compared to the wild type enzyme
V133L
the mutant shows most strongly decreased activity with hemimethylated DNA compared to the wild type enzyme
V133S
the mutant shows strongly decreased activity with hemimethylated DNA compared to the wild type enzyme
Y138A
loses its interaction with the O6 atom of Gua1, shows small changes in specificity factor S1
D181A
-
site-directed mutagenesis, inactive mutant, mutation abolishes base flipping, D181 seems to contact and stabilize the flipped base, i.e. the intermediate state of the base flipping process
H335A
-
the mutant is catalytically inactive and binds to DNA more tightly than the wild type enzyme
K139A
K9A
L122A
-
site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme, DNA binding is similar to the wild-type enzyme
L175P
slightly lowers the ability of the restriction enzyme to cut DNA
N120A
-
site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme, DNA binding is similar to the wild-type enzyme
N120S
-
site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme, DNA binding is similar to the wild-type enzyme
N126A
N132A
the mutant displays increased kcat value compared to the wild type enzyme using the preferred DNA sequence 5'-GCATACCGGATCAAGTAAATG-3'
P134A
-
site-directed mutagenesis, 2-3fold reduced DNA binding compared to the wild-type enzyme
P134G
-
site-directed mutagenesis, 2-3fold reduced DNA binding compared to the wild-type enzyme
R116A
the mutant displays increased kcat value compared to the wild type enzyme using the preferred DNA sequence 5'-GCATACCGGATCAAGTAAATG-3'
R124A
-
site-directed mutagenesis, over 100fold reduced activity and about 10fold reduced DNA binding compared to the wild-type enzyme
R124S/P134S/K139E/F159L/K241E
-
the mutant shows a more than 20fold preference for methylation at GATT, overall corresponding to a 1600fold change in specificity, the mutant is virtually inactive at GATC sites
R137A
R95A
the mutant displays increased kcat value compared to the wild type enzyme using the preferred DNA sequence 5'-GCATACCGGATCAAGTAAATG-3'
S188A
-
site-directed mutagenesis, exchange in the loop next to the active site, 7-8fold reduction of kcat, mutant shows 92% of wild-type enzyme activity
T190A
-
site-directed mutagenesis, mutant shows 75% of wild-type enzyme activity
Y119A
-
site-directed mutagenesis, over 100fold reduced activity compared to the wild-type enzyme, 2-3fold reduced DNA binding compared to the wild-type enzyme
Y138A
-
site-directed mutagenesis, DNA binding is similar to the wild-type enzyme
Y138R
-
the mutant which carries both base Gua1 recognition elements (K9 from EcoDam) is fully active and specific, about 2fold more active than the wild type enzyme
Y184A
-
site-directed mutagenesis, mutant shows 1.7% of wild-type enzyme activity
additional information
L122A
reduces the size of an aliphatic hydrocarbon side chain, is sufficient to convert EcoDam into a bona fide maintenance MTase with pronounced preference for hemimethylated DNA
L122A
the mutant is almost inactive on unmethylated DNA. The variant is able to sense the methylation status of the 5'-GATC-3' double-stranded target recognition site and methylates only hemimethylated DNA with 35-40% of wild type activity
L122A/V133L
the mutant shows increased activity with hemimethylated DNA compared to the wild type enzyme
L122A/V133L
the variant is able to sense the methylation status of the 5'-GATC-3' double-stranded target recognition site and methylates only hemimethylated DNA
P134A
high catalytic activity, exhibits only a small reduction in the amplitude of the fluorescence change, but no detectable changes in the kinetics of base-flipping, induces base-flipping of the substrate with altered sequence at the third base-pair
P134A
the variant methylates hemimethylated DNA with 100% of wild type activity
K139A
-
site-directed mutagenesis, 2-3fold reduced DNA binding compared to the wild-type enzyme
K139A
the mutant displays increased kcat value compared to the wild type enzyme using the preferred DNA sequence 5'-GCATACCGGATCAAGTAAATG-3'
K9A
-
the mutant shows strongly reduced methylation activity towards the sequence GATC
N126A
the mutant displays decreased kcat value compared to the wild type enzyme using the preferred DNA sequence 5'-GCATACCGGATCAAGTAAATG-3'
R137A
-
site-directed mutagenesis, DNA binding is similar to the wild-type enzyme
R137A
the mutant displays increased kcat value compared to the wild type enzyme using the preferred DNA sequence 5'-GCATACCGGATCAAGTAAATG-3'
additional information
EcoDam K9A variant, shows slightly reduced catalytic activity and DNA binding, shows a loss of specificity at the first base-pair, unable to methylate any of the near-cognate sites, base-flipping of substrates carrying a base-pair substitution at the first position of the target site is more efficient than with wild-type
additional information
-
EcoDam K9A variant, shows slightly reduced catalytic activity and DNA binding, shows a loss of specificity at the first base-pair, unable to methylate any of the near-cognate sites, base-flipping of substrates carrying a base-pair substitution at the first position of the target site is more efficient than with wild-type
additional information
-
no methylation of the TGATCprox site after mutation of the site, and subsequently no PapI-dependent binding of Lrp to binding sites 1-3
additional information
-
complementation of the Escherichia coli dam mutant strain GM2163 with the Yersinia enterocolitica dam gene
additional information
-
mutant strains dam, dam mutS, and mutS, deficient in dam and/or mismatch repair, mutS strain does not display many differences from the wild-type at the transcriptional level, both dam and dam mutS strains show differential expression of 206 and 114 genes and expression at higher levels as wild-type, dam mutS strain shows higher variability among some of these induced genes than the dam strain, dam strain has a significantly higher level of basal double-strand breaks than the dam mutS strain
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni2+ affinity column chromatography, UnoS column chromatography, and S75 Sepharose column chromatography
phosphocellulose column chromatography and Blue Sepharose column chromatography
ceramic hydroxylapatite column chromatography and cation exchange column chromatography
-
phosphocellulose column chromatography and Blue Sepharose column chromatography
phosphocellulose column chromatography and Blue Sepharose column chromatography
-
phosphocellulose column chromatography and Blue Sepharose column chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene yhdJ or ccrM, chromosomally encoded, DNA sequence determination and analysis, overvexpression decreases bacterial growth by altering cell division, cells show aberant morphology
-
overproduction in Yersinia enterocolitica, increased spontaneous resistance to chloramphenicol 21.2fold and to streptomycin 39.4fold, decrease in resistance to 2-aminopurine, expression in Yersinia enterocolitica mutant strain GHY121, can complement it
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
the first Gua is recognized by K9, removal of which abrogates the first base-pair recognition, the flipped target Ade binds to the surface of EcoDam in the absence of S-adenosyl-L-methionine, which illustrates a possible intermediate in the base-flipping pathway, the orphaned Thy residue displays structural flexibility by adopting an extrahelical or intrahelical position where it is in contact to N120
analysis
medicine
analysis
-
computational analysis of a published Dam knockout microarray alongside other publicly available
analysis
structural model of the type IC M.EcoR124I DNA methyltransferase, comprising the HsdS subunit, two HsdM subunits, the cofactor S-adenosyl-L-methionine and the substrate DNA molecule
medicine
-
epigenetic effects, in addition to genotoxic effects, need to be considered in chemical carcinogenesis initiated by r7,t8-dihydroxy-t9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene, since the inhibition of methylation may allow the expression of genes that promote tumor development
medicine
-
upregulation of recombinational repair in dam mutants allows for the efficient repair of double-strand breaks whose formation is dependent on functional mismatch repair
medicine
-
DNA adenine methyltransferase identification is a useful technology to map holocarboxylase synthetase binding sites in human chromatin
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Thielking, V.; Du Bois, S.; Eritja, R.; Guschlbauer, W.
Dam methyltransferase from Escherichia coli. Kinetic studies using modified DNA oligomers. Nonmethylated substrates
Biol. Chem.
378
407-415
1997
Escherichia coli
Marzabal, S.; DuBois, S.; Thielking, V.; Cano, A.; Eritja, R.; Guschlbauer, W.
Dam methylase from Escherichia coli: kinetic studies using modified DNA oligomers: hemimethylated substrates
Nucleic Acids Res.
23
3648-3655
1995
Escherichia coli
Jeltsch, A.; Christ, F.; Fatemi, M.; Roth, M.
On the substrate specificity of DNA methyltransferases. Adenine-N6 DNA methyltransferases also modify cytosine residues at position N4
J. Biol. Chem.
274
19538-19544
1999
Escherichia coli, Planomicrobium okeanokoites
Horton, J.R.; Liebert, K.; Hattman, S.; Jeltsch, A.; Cheng, X.
Transition from nonspecific to specific DNA interactions along the substrate-recognition pathway of dam methyltransferase
Cell
121
349-361
2005
Tequatrovirus T4, Escherichia coli
Kossykh, V.G.; Lloyd, R.S.
A DNA adenine methyltransferase of Escherichia coli that is cell cycle regulated and essential for viability
J. Bacteriol.
186
2061-2067
2004
Escherichia coli
Mashhoon, N.; Carroll, M.; Pruss, C.; Eberhard, J.; Ishikawa, S.; Estabrook, R.A.; Reich, N.
Functional characterization of Escherichia coli DNA adenine methyltransferase, a novel target for antibiotics
J. Biol. Chem.
279
52075-52081
2004
Escherichia coli
Liebert, K.; Hermann, A.; Schlickenrieder, M.; Jeltsch, A.
Stopped-flow and mutational analysis of base flipping by the Escherichia coli Dam DNA-(adenine-N6)-methyltransferase
J. Mol. Biol.
341
443-454
2004
Escherichia coli
Hernday, A.D.; Braaten, B.A.; Low, D.A.
The mechanism by which DNA adenine methylase and PapI activate the pap epigenetic switch
Mol. Cell
12
947-957
2003
Escherichia coli
Zinoviev, V.V.; Yakishchik, S.I.; Evdokimov, A.A.; Malygin, E.G.; Hattman, S.
Symmetry elements in DNA structure important for recognition/methylation by DNA [amino]-methyltransferases
Nucleic Acids Res.
32
3930-3934
2004
Tequatrovirus T4, Escherichia coli
Baskunov, V.B.; Subach, F.V.; Kolbanovskiy, A.; Kolbanovskiy, M.; Eremin, S.A.; Johnson, F.; Bonala, R.; Geacintov, N.E.; Gromova, E.S.
Effects of benzo[a]pyrene-deoxyguanosine lesions on DNA methylation catalyzed by EcoRII DNA methyltransferase and on DNA cleavage effected by EcoRII restriction endonuclease
Biochemistry
44
1054-1066
2005
Escherichia coli
Bheemanaik, S.; Reddy, Y.V.; Rao, D.N.
Structure, function and mechanism of exocyclic DNA methyltransferases
Biochem. J.
399
177-190
2006
Enterobacter cloacae, Caulobacter vibrioides, Escherichia coli, Klebsiella pneumoniae, Streptococcus pneumoniae (P04043), Tequatrovirus T4 (P04392), Thermus aquaticus (P14385), Cereibacter sphaeroides (P14751), Moraxella bovis (P23192)
Robbins-Manke, J.L.; Zdraveski, Z.Z.; Marinus, M.; Essigmann, J.M.
Analysis of global gene expression and double-strand-break formation in DNA adenine methyltransferase- and mismatch repair-deficient Escherichia coli
J. Bacteriol.
187
7027-7037
2005
Escherichia coli
Horton, J.R.; Liebert, K.; Bekes, M.; Jeltsch, A.; Cheng, X.
Structure and substrate recognition of the Escherichia coli DNA adenine methyltransferase
J. Mol. Biol.
358
559-570
2006
Escherichia coli (P0AEE8), Escherichia coli
Faelker, S.; Schmidt, M.A.; Heusipp, G.
DNA methylation in Yersinia enterocolitica: role of the DNA adenine methyltransferase in mismatch repair and regulation of virulence factors
Microbiology
151
2291-2299
2005
Escherichia coli, Yersinia enterocolitica
Obarska, A.; Blundell, A.; Feder, M.; Vejsadova, S.; Sisakova, E.; Weiserova, M.; Bujnicki, J.M.; Firman, K.
Structural model for the multisubunit Type IC restriction-modification DNA methyltransferase M.EcoR124I in complex with DNA
Nucleic Acids Res.
34
1992-2005
2006
Escherichia coli (P10484)
Seshasayee, A.S.
An assessment of the role of DNA adenine methyltransferase on gene expression regulation in E. coli
PLoS ONE
2
e273
2007
Escherichia coli
Jois, P.; Madhu, N.; Rao, D.
Role of histidine residues in EcoP15I DNA methyltransferase activity as probed by chemical modification and site-directed mutagenesis
Biochem. J.
410
543-553
2008
Escherichia coli
Coffin, S.R.; Reich, N.O.
Escherichia coli DNA adenine methyltransferase: intrasite processivity and substrate-induced dimerization and activation
Biochemistry
48
7399-7410
2009
Escherichia coli
Kumar, R.; Srivastava, R.; Singh, R.; Surolia, A.; Rao, D.
Activation and inhibition of DNA methyltransferases by S-adenosyl-L-homocysteine analogues
Bioorg. Med. Chem.
16
2276-2285
2008
Escherichia coli
Elsawy, H.; Podobinschi, S.; Chahar, S.; Jeltsch, A.
Transition from EcoDam to T4Dam DNA recognition mechanism without loss of activity and specificity
ChemBioChem
10
2488-2493
2009
Escherichia coli
Low, D.; Casadesus, J.
Clocks and switches: bacterial gene regulation by DNA adenine methylation
Curr. Opin. Microbiol.
11
106-112
2008
Caulobacter vibrioides, Escherichia coli, Salmonella enterica, Yersinia pseudotuberculosis
Marinus, M.G.; Casadesus, J.
Roles of DNA adenine methylation in host-pathogen interactions: mismatch repair, transcriptional regulation, and more
FEMS Microbiol. Rev.
33
488-503
2009
Aggregatibacter actinomycetemcomitans, Aeromonas hydrophila, Brucella abortus, Campylobacter jejuni, Caulobacter vibrioides, Escherichia coli, Salmonella enterica, Klebsiella pneumoniae, Pasteurella multocida, Yersinia enterocolitica, Vibrio cholerae serotype O1
Bang, J.; Bae, S.H.; Park, C.J.; Lee, J.H.; Choi, B.S.
Structural and dynamics study of DNA dodecamer duplexes that contain un-, hemi-, or fully methylated GATC sites
J. Am. Chem. Soc.
130
17688-17696
2008
Escherichia coli
Coffin, S.R.; Reich, N.O.
Modulation of Escherichia coli DNA methyltransferase activity by biologically derived GATC-flanking sequences
J. Biol. Chem.
283
20106-20116
2008
Escherichia coli (P0AEE8), Escherichia coli
Coffin, S.R.; Reich, N.O.
Escherichia coli DNA adenine methyltransferase: the structural basis of processive catalysis and indirect read-out
J. Biol. Chem.
284
18390-18400
2009
Escherichia coli (P0AEE9)
Peterson, S.N.; Reich, N.O.
Competitive Lrp and Dam assembly at the pap regulatory region: implications for mechanisms of epigenetic regulation
J. Mol. Biol.
383
92-105
2008
Escherichia coli
Chahar, S.; Elsawy, H.; Ragozin, S.; Jeltsch, A.
Changing the DNA recognition specificity of the EcoDam DNA-(adenine-N6)-methyltransferase by directed evolution
J. Mol. Biol.
395
79-88
2010
Escherichia coli
Singh, D.; Pannier, A.; Zempleni, J.
Identification of holocarboxylase synthetase chromatin binding sites in human mammary cell lines using the DNA adenine methyltransferase identification technology
Anal. Biochem.
413
55-59
2011
Escherichia coli
Bonnist, E.Y.; Liebert, K.; Dryden, D.T.; Jeltsch, A.; Jones, A.C.
Using the fluorescence decay of 2-aminopurine to investigate conformational change in the recognition sequence of the EcoRV DNA-(adenine-N6)-methyltransferase on enzyme binding
Biophys. Chem.
160
28-34
2012
Escherichia coli
Pollak, A.J.; Reich, N.O.
Proximal recognition sites facilitate intrasite hopping by DNA adenine methyltransferase: Mechanistic exploration of epigenetic gene regulation
J. Biol. Chem.
287
22873-22881
2012
Escherichia coli
Clark, T.A.; Murray, I.A.; Morgan, R.D.; Kislyuk, A.O.; Spittle, K.E.; Boitano, M.; Fomenkov, A.; Roberts, R.J.; Korlach, J.
Characterization of DNA methyltransferase specificities using single-molecule, real-time DNA sequencing
Nucleic Acids Res.
40
e29
2012
Escherichia coli
Pollak, A.; Reich, N.
DNA adenine methyltransferase facilitated diffusion is enhanced by protein-DNA roadblock complexes that induce DNA looping
Biochemistry
54
2181-2192
2015
Escherichia coli
Elsawy, H.; Chahar, S.
Increasing DNA substrate specificity of the EcoDam DNA-(adenine N6)-methyltransferase by site-directed mutagenesis
Biochemistry
79
1262-1266
2014
Escherichia coli (P0AEE8)
Horton, J.; Zhang, X.; Blumenthal, R.; Cheng, X.
Structures of Escherichia coli DNA adenine methyltransferase (Dam) in complex with a non-GATC sequence: Potential implications for methylation-independent transcriptional repression
Nucleic Acids Res.
43
4296-4308
2015
Escherichia coli (P0AEE8), Escherichia coli
Wons, E.; Mruk, I.; Kaczorowski, T.
Relaxed specificity of prokaryotic DNA methyltransferases results in DNA site-specific modification of RNA/DNA heteroduplexes
J. Appl. Genet.
56
539-546
2015
Escherichia coli, Lactococcus cremoris, Lactococcus cremoris W15, Escherichia coli E1585-68
Gupta, Y.K.; Chan, S.H.; Xu, S.Y.; Aggarwal, A.K.
Structural basis of asymmetric DNA methylation and ATP-triggered long-range diffusion by EcoP15I
Nat. Commun.
6
7363
2015
Caulobacter vibrioides, Escherichia coli
Wons, E.; Mruk, I.; Kaczorowski, T.
Isospecific adenine DNA methyltransferases show distinct preferences towards DNA substrates
Sci. Rep.
8
8243
2018
Escherichia coli, Haemophilus influenzae, Lactococcus cremoris, Haemophilus influenzae RD, Lactococcus cremoris W15, Escherichia coli E1585-68
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