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Information on EC 2.1.1.230 - 23S rRNA (adenosine1067-2'-O)-methyltransferase and Organism(s) Streptomyces azureus and UniProt Accession P18644
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2.1.1.230 23S rRNA (adenosine1067-2'-O)-methyltransferaseIUBMB Comments
The methylase that is responsible for autoimmunity in the thiostrepton producer Streptomyces azureus, renders ribosomes completely resistant to thiostrepton .
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The taxonomic range for the selected organisms is: Streptomyces azureus
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Reaction Schemes
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adenosine1067 in 23S rRNA
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2'-O-methyladenosine1067 in 23S rRNA
Synonyms
nosiheptide-resistance methyltransferase, thiostrepton-resistance methyltransferase, nhr protein, nosiheptide resistance methyltransferase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:23S rRNA (adenosine1067-2'-O)-methyltransferase
The methylase that is responsible for autoimmunity in the thiostrepton producer Streptomyces azureus, renders ribosomes completely resistant to thiostrepton [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 + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
S-adenosyl-L-methionine + adenosine in an RNA fragment containing nucleotides 1029-1122 of the 23S ribosomal RNA from Escherichia coli
S-adenosyl-L-homocysteine + 2'-O-methyladenosine in an RNA fragment containing nucleotides 1029-1122 of the 23S ribosomal RNA from Escherichia coli
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-
-
-
?
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
enzyme Tsr uses the co-substrate AdoMet to methylate the 23 S rRNA, presumably prior to the assembly of the 50 S subunit as the L11 and proposed Tsr binding surfaces are overlapping
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-
?
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
58-nt RNA substrate secondary structure, and key longrange interactions within the 58-nt domain tertiary fold, overview
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-
?
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
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-
-
-
?
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
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Streptomyces azureus is the producer of the peptide antibiotic thiostrepton. Thiostrepton inhibits protein synthesis by binding to the complex of 23S rRNA and protein L-11 which blocks processes associated with the GTP-hydrolysis center of the ribosome including the binding of guanine nucleotides, elongation factor proteins and tRNA. 23S rRNA (adenosine1067-2'-O)-methyltransferase confers resistance to tthiostrepton
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-
?
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
-
the enzyme is involved in resistance to thiostrepton
-
-
?
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
-
the methylase enzyme, responsible for autoimmunity in the thiostrepton producer Streptomyces azureus, renders ribosomes completely resistant to thiostrepton
-
-
?
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
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RNA sequence variants of the RNA fragment with mutations in nucleotides 1051-1108 are tested as substrates for the methylase. Methylation is dependent on the secondary structure of the hairpin including nucleotide A1067 and the exact sequence U(1066)-A(1067)-G(1068)-A(1O69)-A(1070) of the single strand
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?
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 + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
enzyme Tsr uses the co-substrate AdoMet to methylate the 23 S rRNA, presumably prior to the assembly of the 50 S subunit as the L11 and proposed Tsr binding surfaces are overlapping
-
-
?
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
-
Streptomyces azureus is the producer of the peptide antibiotic thiostrepton. Thiostrepton inhibits protein synthesis by binding to the complex of 23S rRNA and protein L-11 which blocks processes associated with the GTP-hydrolysis center of the ribosome including the binding of guanine nucleotides, elongation factor proteins and tRNA. 23S rRNA (adenosine1067-2'-O)-methyltransferase confers resistance to tthiostrepton
-
-
?
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
-
the enzyme is involved in resistance to thiostrepton
-
-
?
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA
-
the methylase enzyme, responsible for autoimmunity in the thiostrepton producer Streptomyces azureus, renders ribosomes completely resistant to thiostrepton
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-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Protein L11
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binding of either thiostrepton or protein L11 inhibits methylation
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S-Adenosyl-D-homocysteine
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competitivewith respect to S-adenosyl-L-methionine
Thiostrepton
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binding of either thiostrepton or protein L11 inhibits methylation
additional information
stabilizing the RNA tertiary structure, as in RNA mutant U1061A, decreases Tsr binding affinity and catalytic activity independent of RNA structural changes
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additional information
-
stabilizing the RNA tertiary structure, as in RNA mutant U1061A, decreases Tsr binding affinity and catalytic activity independent of RNA structural changes
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.001
adenosine in an RNA fragment containing nucleotides 1029-1122 of the 23S ribosomal RNA from Escherichia coli
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pH 7.5, 35°C
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7 - 7.8
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pH 7.0: about 60% of maximal activity, pH 7.8: about 60% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
in the absence of the N-terminal domain, Tsr is catalytically inactive despite possessing the intact S-adenosyl-L-methionine binding sites and catalytic center. The Tsr-C-terminal domain dimer binds the RNA 30fold more weakly than the wild-type enzyme and is unable to promote the N-terminal domain-dependent RNA conformational change
physiological function
the thiostrepton producer Streptomyces azureus prevents self-intoxication by expressing the thiostrepton-resistance methyltransferase (Tsr), which methylates the 2'-hydroxyl of 23 S rRNA nucleotide adenosine 1067 within the thiostrepton binding site
physiological function
additional information
physiological function
-
Streptomyces azureus is the producer of the peptide antibiotic thiostrepton. Thiostrepton inhibits protein synthesis by binding to the complex of 23S rRNA and protein L-11 which blocks processes associated with the GTP-hydrolysis center of the ribosome including the binding of guanine nucleotides, elongation factor proteins and tRNA. 23S rRNA (adenosine1067-2'-O)-methyltransferase confers resistance to thiostrepton
physiological function
-
the enzyme is involved in resistance to thiostrepton
physiological function
-
the methylase enzyme, responsible for autoimmunity in the thiostrepton producer Streptomyces azureus, renders ribosomes completely resistant to thiostrepton
additional information
each protomer of the homodimer containing an L30e-like amino-terminal domain and a SPOUT methyltransferase family catalytic carboxyl-terminal domain, both enzyme domains are required for high affinity RNA substrate binding. The Tsr-C-terminal domain has intrinsic, weak RNA affinity that is necessary to direct the specific high-affinity Tsr-RNA interaction via N-terminal domains, which have no detectable RNA affinity when isolated. The N-terminal domains increase RNA binding affinity and are necessary for catalysis, RNA binding mechanism, overview. The N-terminal domain of Tsr is an essential component of the RNA substrate recognition mechanism by both promoting high affinity RNA binding and activation of catalysis by the C-terminal domain
additional information
-
each protomer of the homodimer containing an L30e-like amino-terminal domain and a SPOUT methyltransferase family catalytic carboxyl-terminal domain, both enzyme domains are required for high affinity RNA substrate binding. The Tsr-C-terminal domain has intrinsic, weak RNA affinity that is necessary to direct the specific high-affinity Tsr-RNA interaction via N-terminal domains, which have no detectable RNA affinity when isolated. The N-terminal domains increase RNA binding affinity and are necessary for catalysis, RNA binding mechanism, overview. The N-terminal domain of Tsr is an essential component of the RNA substrate recognition mechanism by both promoting high affinity RNA binding and activation of catalysis by the C-terminal domain
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). TSNR_STRAJ
269
0
28901
Swiss-Prot
other Location (Reliability: 4)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homodimer
each protomer containing an L30e-like amino-terminal domain and a SPOUT methyltransferase family catalytic carboxyl-terminal domain, both enzyme domains are required for high affinity RNA substrate binding, Tsr domain organization, overview
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
R162A
site-directed mutagenesis, the mutant binds RNA with 100fold weaker affinity than wild-type Tsr
R26A
site-directed mutagenesis, the mutant is unable to promote the RNase V1-sensitive RNA structural changes, but maintains its interaction with the RNA under certain conditions for the protection of nucleotides G1068 and G1071 from cleavage by RNase T1
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)-pLysS by nickel affinity and heparin affinity chromatography, followed by gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene tsr, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)-pLysS
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Thompson, J.; Cundliffe, E.
Purification and properties of an RNA methylase produced by Streptomyces azureus and involved in resistance to thiostrepton
J. Gen. Microbiol.
124
291-297
1981
Streptomyces azureus
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Thompson, J.; Schmidt, F.; Cundliffe, E.
Site of action of a ribosomal RNA methylase conferring resistance to thiostrepton
J. Biol. Chem.
257
7915-7917
1982
Streptomyces azureus
Bechthold, A.; Floss, H.G.
Overexpression of the thiostrepton-resistance gene from Streptomyces azureus in Escherichia coli and characterization of recognition sites of the 23S rRNA A1067 2'-methyltransferase in the guanosine triphosphatase center of 23S ribosomal RNA
Eur. J. Biochem.
224
431-437
1994
Streptomyces azureus
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