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show all sequences of 2.1.1.170

Structural and functional studies of the Thermus thermophilus 16S rRNA methyltransferase RsmG

Gregory, S.T.; Demirci, H.; Belardinelli, R.; Monshupanee, T.; Gualerzi, C.; Dahlberg, A.E.; Jogl, G.; RNA 15, 1693-1704 (2009)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
-
Thermus thermophilus
Crystallization (Commentary)
Crystallization
Organism
microbatch technique under oil at 4°C. Determination of the structure of RsmG (249 amino acids) in three different crystal forms: the enzyme in complex with the cofactor S-adensosyl-L-methionine, the enzyme in complex with S-adenosyl-L-homocysteine, the enzyme in complex with adenosine monophosphate and S-adenosyl-L-methionine. RsmG X-ray crystal structures at up to 1.5 A resolution. Cofactor-bound crystal structures of RsmG reveals a positively charged surface area remote from the active site that binds an adenosine monophosphate molecule
Thermus thermophilus
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
the 30S subunits in their native conformation are not a proper substrate and removal of Mg2+ ions from the subunit is required to open the structure sufficiently to expose elements involved in enzyme binding
Thermus thermophilus
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
S-adenosyl-L-methionine + guanine527 in 16S rRNA
Thermus thermophilus
methylations concentrated in the decoding site of the 30S ribosomal subunit may act to fine tune codon recognition in a manner similar to tRNA modifications. The intact 30S subunit is very unlikely to be the natural substrate for Thermus thermophilus RsmG in vivo. This interpretation is consistent with the position of G527 in the intact 30S subunit, where it is buried and would be inaccessible for methylation without substantial unfolding of the local subunit structure. Deproteinized 16S rRNA is the most active substrate in vitro. In vivo, several ribosomal proteins probably begin binding to the nascent 16S rRNA transcript prior to its completion, making an early assembly intermediate a plausible candidate for the biological substrate of RsmG
S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA
-
-
?
S-adenosyl-L-methionine + guanine527 in 16S rRNA
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
methylations concentrated in the decoding site of the 30S ribosomal subunit may act to fine tune codon recognition in a manner similar to tRNA modifications. The intact 30S subunit is very unlikely to be the natural substrate for Thermus thermophilus RsmG in vivo. This interpretation is consistent with the position of G527 in the intact 30S subunit, where it is buried and would be inaccessible for methylation without substantial unfolding of the local subunit structure. Deproteinized 16S rRNA is the most active substrate in vitro. In vivo, several ribosomal proteins probably begin binding to the nascent 16S rRNA transcript prior to its completion, making an early assembly intermediate a plausible candidate for the biological substrate of RsmG
S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Thermus thermophilus
Q9LCY2
-
-
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
Q9LCY2
-
-
Purification (Commentary)
Commentary
Organism
the AdoMet cofactor is tightly bound in RsmG and copurifies with the recombinant protein
Thermus thermophilus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
S-adenosyl-L-methionine + guanine527 in 16S rRNA
methylations concentrated in the decoding site of the 30S ribosomal subunit may act to fine tune codon recognition in a manner similar to tRNA modifications. The intact 30S subunit is very unlikely to be the natural substrate for Thermus thermophilus RsmG in vivo. This interpretation is consistent with the position of G527 in the intact 30S subunit, where it is buried and would be inaccessible for methylation without substantial unfolding of the local subunit structure. Deproteinized 16S rRNA is the most active substrate in vitro. In vivo, several ribosomal proteins probably begin binding to the nascent 16S rRNA transcript prior to its completion, making an early assembly intermediate a plausible candidate for the biological substrate of RsmG
706725
Thermus thermophilus
S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA
-
-
-
?
S-adenosyl-L-methionine + guanine527 in 16S rRNA
the most active substrate for Thermus thermophilus RsmG in vitro is deproteinized 16S rRNA. 30S subunits in their native conformation are not a proper substrate, removal of Mg2+ ions from the subunit is required to open the structure sufficiently to expose elements involved in enzyme binding. Identification of methylated nucleotide
706725
Thermus thermophilus
S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA
-
-
-
?
S-adenosyl-L-methionine + guanine527 in 16S rRNA
methylations concentrated in the decoding site of the 30S ribosomal subunit may act to fine tune codon recognition in a manner similar to tRNA modifications. The intact 30S subunit is very unlikely to be the natural substrate for Thermus thermophilus RsmG in vivo. This interpretation is consistent with the position of G527 in the intact 30S subunit, where it is buried and would be inaccessible for methylation without substantial unfolding of the local subunit structure. Deproteinized 16S rRNA is the most active substrate in vitro. In vivo, several ribosomal proteins probably begin binding to the nascent 16S rRNA transcript prior to its completion, making an early assembly intermediate a plausible candidate for the biological substrate of RsmG
706725
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA
-
-
-
?
S-adenosyl-L-methionine + guanine527 in 16S rRNA
the most active substrate for Thermus thermophilus RsmG in vitro is deproteinized 16S rRNA. 30S subunits in their native conformation are not a proper substrate, removal of Mg2+ ions from the subunit is required to open the structure sufficiently to expose elements involved in enzyme binding. Identification of methylated nucleotide
706725
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA
-
-
-
?
Cofactor
Cofactor
Commentary
Organism
Structure
S-adenosyl-L-methionine
S-adenosyl-L-methionine is bound in a canonical conformation above the beta-sheet and close to the conserved GxGxG methyltransferase signature motif (residues 88–92 between strand beta1 and helix alpha4). The AdoMet cofactor is tightly bound in RsmG and copurifies with the recombinant protein
Thermus thermophilus
Cloned(Commentary) (protein specific)
Commentary
Organism
-
Thermus thermophilus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
S-adenosyl-L-methionine
S-adenosyl-L-methionine is bound in a canonical conformation above the beta-sheet and close to the conserved GxGxG methyltransferase signature motif (residues 88–92 between strand beta1 and helix alpha4). The AdoMet cofactor is tightly bound in RsmG and copurifies with the recombinant protein
Thermus thermophilus
Crystallization (Commentary) (protein specific)
Crystallization
Organism
microbatch technique under oil at 4°C. Determination of the structure of RsmG (249 amino acids) in three different crystal forms: the enzyme in complex with the cofactor S-adensosyl-L-methionine, the enzyme in complex with S-adenosyl-L-homocysteine, the enzyme in complex with adenosine monophosphate and S-adenosyl-L-methionine. RsmG X-ray crystal structures at up to 1.5 A resolution. Cofactor-bound crystal structures of RsmG reveals a positively charged surface area remote from the active site that binds an adenosine monophosphate molecule
Thermus thermophilus
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
the 30S subunits in their native conformation are not a proper substrate and removal of Mg2+ ions from the subunit is required to open the structure sufficiently to expose elements involved in enzyme binding
Thermus thermophilus
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
S-adenosyl-L-methionine + guanine527 in 16S rRNA
Thermus thermophilus
methylations concentrated in the decoding site of the 30S ribosomal subunit may act to fine tune codon recognition in a manner similar to tRNA modifications. The intact 30S subunit is very unlikely to be the natural substrate for Thermus thermophilus RsmG in vivo. This interpretation is consistent with the position of G527 in the intact 30S subunit, where it is buried and would be inaccessible for methylation without substantial unfolding of the local subunit structure. Deproteinized 16S rRNA is the most active substrate in vitro. In vivo, several ribosomal proteins probably begin binding to the nascent 16S rRNA transcript prior to its completion, making an early assembly intermediate a plausible candidate for the biological substrate of RsmG
S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA
-
-
?
S-adenosyl-L-methionine + guanine527 in 16S rRNA
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
methylations concentrated in the decoding site of the 30S ribosomal subunit may act to fine tune codon recognition in a manner similar to tRNA modifications. The intact 30S subunit is very unlikely to be the natural substrate for Thermus thermophilus RsmG in vivo. This interpretation is consistent with the position of G527 in the intact 30S subunit, where it is buried and would be inaccessible for methylation without substantial unfolding of the local subunit structure. Deproteinized 16S rRNA is the most active substrate in vitro. In vivo, several ribosomal proteins probably begin binding to the nascent 16S rRNA transcript prior to its completion, making an early assembly intermediate a plausible candidate for the biological substrate of RsmG
S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
the AdoMet cofactor is tightly bound in RsmG and copurifies with the recombinant protein
Thermus thermophilus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
S-adenosyl-L-methionine + guanine527 in 16S rRNA
methylations concentrated in the decoding site of the 30S ribosomal subunit may act to fine tune codon recognition in a manner similar to tRNA modifications. The intact 30S subunit is very unlikely to be the natural substrate for Thermus thermophilus RsmG in vivo. This interpretation is consistent with the position of G527 in the intact 30S subunit, where it is buried and would be inaccessible for methylation without substantial unfolding of the local subunit structure. Deproteinized 16S rRNA is the most active substrate in vitro. In vivo, several ribosomal proteins probably begin binding to the nascent 16S rRNA transcript prior to its completion, making an early assembly intermediate a plausible candidate for the biological substrate of RsmG
706725
Thermus thermophilus
S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA
-
-
-
?
S-adenosyl-L-methionine + guanine527 in 16S rRNA
the most active substrate for Thermus thermophilus RsmG in vitro is deproteinized 16S rRNA. 30S subunits in their native conformation are not a proper substrate, removal of Mg2+ ions from the subunit is required to open the structure sufficiently to expose elements involved in enzyme binding. Identification of methylated nucleotide
706725
Thermus thermophilus
S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA
-
-
-
?
S-adenosyl-L-methionine + guanine527 in 16S rRNA
methylations concentrated in the decoding site of the 30S ribosomal subunit may act to fine tune codon recognition in a manner similar to tRNA modifications. The intact 30S subunit is very unlikely to be the natural substrate for Thermus thermophilus RsmG in vivo. This interpretation is consistent with the position of G527 in the intact 30S subunit, where it is buried and would be inaccessible for methylation without substantial unfolding of the local subunit structure. Deproteinized 16S rRNA is the most active substrate in vitro. In vivo, several ribosomal proteins probably begin binding to the nascent 16S rRNA transcript prior to its completion, making an early assembly intermediate a plausible candidate for the biological substrate of RsmG
706725
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA
-
-
-
?
S-adenosyl-L-methionine + guanine527 in 16S rRNA
the most active substrate for Thermus thermophilus RsmG in vitro is deproteinized 16S rRNA. 30S subunits in their native conformation are not a proper substrate, removal of Mg2+ ions from the subunit is required to open the structure sufficiently to expose elements involved in enzyme binding. Identification of methylated nucleotide
706725
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA
-
-
-
?
General Information
General Information
Commentary
Organism
malfunction
construction of an rsmG null allele by deleting the rsmG coding sequence and replacing it with htk, encoding a heat-stable kanamycin adenyltransferase. This null allele retains the very N- and C-terminal rsmG coding sequences, in-frame with the htk coding sequence, in order to maintain the rsmG–parA overlap and minimize any effects on parA expression. This allele is designated DrsmGThtk2 and the mutant containing this allele is designated HG 917. Thermus thermophilus rsmG mutants are weakly resistant to the aminoglycoside antibiotic streptomycin. Growth competition experiments indicate a physiological cost to loss of RsmG activity, consistent with the conservation of the modification site in the decoding region of the ribosome
Thermus thermophilus
General Information (protein specific)
General Information
Commentary
Organism
malfunction
construction of an rsmG null allele by deleting the rsmG coding sequence and replacing it with htk, encoding a heat-stable kanamycin adenyltransferase. This null allele retains the very N- and C-terminal rsmG coding sequences, in-frame with the htk coding sequence, in order to maintain the rsmG–parA overlap and minimize any effects on parA expression. This allele is designated DrsmGThtk2 and the mutant containing this allele is designated HG 917. Thermus thermophilus rsmG mutants are weakly resistant to the aminoglycoside antibiotic streptomycin. Growth competition experiments indicate a physiological cost to loss of RsmG activity, consistent with the conservation of the modification site in the decoding region of the ribosome
Thermus thermophilus
Other publictions for EC 2.1.1.170
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
733472
Benitez-Paez
Impairing methylations at ribo ...
Escherichia coli
Biomedica
34
41-49
2014
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1
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2
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1
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1
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1
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1
1
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721042
Benitez-Paez
Regulation of expression and c ...
Escherichia coli
RNA
18
795-806
2012
1
-
1
-
15
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2
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1
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1
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1
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1
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1
1
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15
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1
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1
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1
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2
3
3
2
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-
704222
Tanaka
Activation of secondary metabo ...
Streptomyces griseus, Streptomyces griseus IFO13189
J. Antibiot.
62
669-673
2009
-
-
-
-
-
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2
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2
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1
1
-
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706725
Gregory
Structural and functional stud ...
Thermus thermophilus, Thermus thermophilus HB8 / ATCC 27634 / DSM 579
RNA
15
1693-1704
2009
-
-
1
1
-
-
-
-
-
1
-
2
-
5
-
-
1
-
-
-
-
-
4
-
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-
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-
1
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1
1
1
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1
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2
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1
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-
4
-
-
-
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-
-
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1
1
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-
704272
Nishimura
Mutations in rsmG, encoding a ...
Streptomyces coelicolor, Streptomyces coelicolor A3(2)
J. Bacteriol.
189
3876-3883
2007
-
-
-
-
-
-
-
-
-
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6
-
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-
-
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-
-
-
-
1
1
-
-
-
704273
Nishimura
Identification of the RsmG met ...
Bacillus subtilis 168, Bacillus subtilis
J. Bacteriol.
189
6068-6073
2007
-
-
-
-
-
-
-
-
-
-
-
2
-
90
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4
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2
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4
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1
1
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705767
Okamoto
Loss of a conserved 7-methylgu ...
Escherichia coli, Escherichia coli BW25113
Mol. Microbiol.
63
1096-1106
2007
-
-
1
-
1
-
-
-
-
-
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28
-
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1
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2
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1
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1
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1
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1
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1
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2
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1
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1
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1
1
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706646
Romanowski
Crystal structure of the Esche ...
Escherichia coli
Proteins
47
563-567
2002
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1
1
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2
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2
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1
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1
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