Literature summary for 2.1.1.170 extracted from

Gregory, S.T.; Demirci, H.; Belardinelli, R.; Monshupanee, T.; Gualerzi, C.; Dahlberg, A.E.; Jogl, G.
Structural and functional studies of the Thermus thermophilus 16S rRNA methyltransferase RsmG (2009), RNA, 15, 1693-1704.

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Cloned(Commentary)

Cloned (Comment)	Organism
-	Thermus thermophilus

Crystallization (Commentary)

Crystallization (Comment)	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

Crystallization (Comment)

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	Comment	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	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
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	UniProt	Comment	Textmining
Thermus thermophilus	Q9LCY2	-	-
Thermus thermophilus HB8 / ATCC 27634 / DSM 579	Q9LCY2	-	-

Purification (Commentary)

Purification (Comment)	Organism
the AdoMet cofactor is tightly bound in RsmG and copurifies with the recombinant protein	Thermus thermophilus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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	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	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	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	Thermus thermophilus HB8 / ATCC 27634 / DSM 579	S-adenosyl-L-homocysteine + N7-methylguanine527 in 16S rRNA	-	?

Synonyms

Synonyms	Comment	Organism
16S rRNA methyltransferase RsmG	-	Thermus thermophilus
gidB	-	Thermus thermophilus
ribosomal RNA small subunit methyltransferase G	-	Thermus thermophilus
rsmG	-	Thermus thermophilus

Cofactor

Cofactor	Comment	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 8892 between strand beta1 and helix alpha4). The AdoMet cofactor is tightly bound in RsmG and copurifies with the recombinant protein	Thermus thermophilus

General Information

General Information	Comment	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 rsmGparA 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