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S-adenosyl-L-methionine + cytosine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 5-methylcytosine1405 in 16S rRNA
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S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + N7-methylguanine1405 in 16S rRNA
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S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
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S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
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S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
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S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
the enzyme confers resistance to aminoglycosides like gentamicin and sisomicin by specifically methylating G1405 in bacterial 16S rRNA
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S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
the enzyme encodes an enzyme that modifies 16S rRNA and thereby confers resistance to 4,6-disubstituted deoxystreptamine aminoglycosides
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S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
the enzyme produced by the antibiotic-producing bacterium Micromonospora zionensis methylates guanine1405 in 16S rRNA to 7-methylguanine, thereby rendering the ribosome resistant to 4,6-disubstituted deoxystreptamine aminoglycosides, which include gentamicins and kanamycins
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S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
methylation site is experimentally determined as guanine1405 by MALDI-ToF mass spectrometry
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S-adenosyl-L-methionine + cytosine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 5-methylcytosine1405 in 16S rRNA
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?
S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + N7-methylguanine1405 in 16S rRNA
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?
S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
-
-
-
?
S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
the enzyme confers resistance to aminoglycosides like gentamicin and sisomicin by specifically methylating G1405 in bacterial 16S rRNA
-
-
?
S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
the enzyme encodes an enzyme that modifies 16S rRNA and thereby confers resistance to 4,6-disubstituted deoxystreptamine aminoglycosides
-
-
?
S-adenosyl-L-methionine + guanine1405 in 16S rRNA
S-adenosyl-L-homocysteine + 7-methylguanine1405 in 16S rRNA
the enzyme produced by the antibiotic-producing bacterium Micromonospora zionensis methylates guanine1405 in 16S rRNA to 7-methylguanine, thereby rendering the ribosome resistant to 4,6-disubstituted deoxystreptamine aminoglycosides, which include gentamicins and kanamycins
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additional information
Sgm shows intrinsic N7-G1405 16S-RMTase activity
physiological function
encodes an enzyme that modifies 16S rRNA and thereby confers resistance to 4,6-disubstituted deoxystreptamine aminoglycosides. The expression of the sgm gene is regulated by the translational autorepression
physiological function
the enzyme produced by the antibiotic-producing bacterium Micromonospora zionensis methylates guanine1405 in 16S rRNA to 7-methylguanine, thereby rendering the ribosome resistant to 4,6-disubstituted deoxystreptamine aminoglycosides, which include gentamicins and kanamycins
physiological function
most aminoglycosides bind to the decoding aminoacyl-tRNA recognition site (A-site) of the 16S rRNA that composes bacterial 30S ribosome, and subsequently interfere with bacterial growth through blocking of protein synthesis. These aminoglycoside-producing actinomycetes are inherently resistant to aminoglycosides, because they harbor intrinsic 16S rRNA methyltransferase genes, that can confer aminoglycoside resistance to bacteria by modifying specific nucleotide residues in the aminoglycoside binding site of 16S rRNA. Aminoglycoside resistance profile provided by N7-G1405 16S-RMTases, overview
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D158A
mutant with drastically increased sensitivity to kanamycin
K54A
mutant with drastically increased sensitivity to kanamycin
R108A
mutant with drastically increased sensitivity to kanamycin
R187S
mutant with drastically increased sensitivity to kanamycin
R187S/G212S
mutant with strongly reduced activity
R433A
mutant with drastically increased sensitivity to kanamycin
additional information
analysis of sequencefunction relationships of Sgm MTase by limited proteolysis and site-directed and random mutagenesis
D156A
mutant with drastically increased sensitivity to kanamycin
D156A
no binding of S-adenosyl-L-methionine
D182A
mutant with drastically increased sensitivity to kanamycin
D182A
no binding of S-adenosyl-L-methionine
E205A
mutant retains S-adenosyl-L-methionine binding
E205A
mutant with drastically increased sensitivity to kanamycin
E267A
mutant retains S-adenosyl-L-methionine binding
E267A
mutant with drastically increased sensitivity to kanamycin
G135A
mutant with drastically increased sensitivity to kanamycin
G135A
no binding of S-adenosyl-L-methionine
K199A
mutant retains S-adenosyl-L-methionine binding
K199A
mutant with drastically increased sensitivity to kanamycin
R236A
mutant retains S-adenosyl-L-methionine binding
R236A
mutant with drastically increased sensitivity to kanamycin
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cloned in Streptomyces lividans. The sgm gene is expressed in Micromonospora melanosporea, where its own promoter is active, and also in Escherichia coli under the control of the lacZ promoter
cloning of the sgm gene into pET-25b (+) vector with the addition of N-terminal non-cleavable His6 tag, as well as alanine mutagenesis of residues D156, D182 and R108. The constructs are co-transformed along with pGroESL into the strain BL21 (DE3) of Escherichia coli for protein expression
expression in Escherichia coli
gene sgm, phylogenetic analysis, sequence comparisons
introduction of a recombinant version of the Sgm methyltransferase gene from Micromonospora zionensis into an Escherichia coli strain that has a full complement of housekeeping methyltransferases. Analyses of the 16S rRNA shows that the m5C1407-specific housekeeping methyltransferase RsmF (YebU) is outcompeted by Sgm as the resistance methyltransferase gains access to its own m7G1405 target on the 30S ribosomal subunit. A single amino acid change in Sgm, which lowers the level of conferred resistance, reduces the ability of Sgm to interfere with RsmF methylation on the 30S subunit
sgm is cloned into a yeast expression vector to test whether the prokaryotic methylases can modify the 18S rRNA A-site and thus confer resistance to the aminoglycoside antibiotic G-418. Sgm does not provide resistant phenotypes to yeast cells. Despite all similarities in the antibiotic binding site, methylation by Sgm does not occur in yeast, suggesting that the recognition site for these methylases could be different in 30S and 40S subunits
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Maravic Vlahovicek, G.; Cubrilo, S.; Tkaczuk, K.L.; Bujnicki, J.M.
Modeling and experimental analyses reveal a two-domain structure and amino acids important for the activity of aminoglycoside resistance methyltransferase Sgm
Biochim. Biophys. Acta
1784
582-590
2008
Micromonospora zionensis (Q7M0R2)
brenda
Kojic, M.; Topisirovic, L.; Vasiljevic, B.
Cloning and characterization of an aminoglycoside resistance determinant from Micromonospora zionensis
J. Bacteriol.
174
7868-7872
1992
Micromonospora zionensis (Q7M0R2), Micromonospora zionensis
brenda
Kojic, M.; Topisirovic, L.; Vasiljevic, B.
Translational autoregulation of the sgm gene from Micromonospora zionensis
J. Bacteriol.
178
5493-5498
1996
Micromonospora zionensis (Q7M0R2), Micromonospora zionensis
brenda
Savic, M.; Ilic-Tomic, T.; Macmaster, R.; Vasiljevic, B.; Conn, G.L.
Critical residues for cofactor binding and catalytic activity in the aminoglycoside resistance methyltransferase Sgm
J. Bacteriol.
190
5855-5861
2008
Micromonospora zionensis (Q7M0R2)
brenda
Savic, M.; Lovric, J.; Tomic, T.I.; Vasiljevic, B.; Conn, G.L.
Determination of the target nucleosides for members of two families of 16S rRNA methyltransferases that confer resistance to partially overlapping groups of aminoglycoside antibiotics
Nucleic Acids Res.
37
5420-5431
2009
Frankia sp. (Q2J7L5), Frankia sp. CcI3 (Q2J7L5), Micromonospora echinospora (Q70KC8), Micromonospora zionensis (Q7M0R2)
brenda
Husain, N.; Tkaczuk, K.L.; Tulsidas, S.R.; Kaminska, K.H.; Cubrilo, S.; Maravic-Vlahovicek, G.; Bujnicki, J.M.; Sivaraman, J.
Structural basis for the methylation of G1405 in 16S rRNA by aminoglycoside resistance methyltransferase Sgm from an antibiotic producer: a diversity of active sites in m7G methyltransferases
Nucleic Acids Res.
38
4120-4132
2010
Micromonospora zionensis (Q7M0R2), Micromonospora zionensis
brenda
Tomic, T.I.; Moric, I.; Conn, G.L.; Vasiljevic, B.
Aminoglycoside resistance genes sgm and kgmB protect bacterial but not yeast small ribosomal subunits in vitro despite high conservation of the rRNA A-site
Res. Microbiol.
159
658-662
2008
Micromonospora zionensis (Q7M0R2)
brenda
Cubrilo, S.; Babic, F.; Douthwaite, S.; Maravic Vlahovicek, G.
The aminoglycoside resistance methyltransferase Sgm impedes RsmF methylation at an adjacent rRNA nucleotide in the ribosomal A site
RNA
15
1492-1497
2009
Micromonospora zionensis (Q7M0R2)
brenda
Wachino, J.; Arakawa, Y.
Exogenously acquired 16S rRNA methyltransferases found in aminoglycoside-resistant pathogenic Gram-negative bacteria: an update
Drug Resist. Updat.
15
133-148
2012
Citrobacter freundii (E9N3S1), Micromonospora echinospora (Q70KC8), Micromonospora inyonensis (Q5Y813), Micromonospora olivasterospora (Q08325), Micromonospora rosea (P24619), Micromonospora zionensis (Q7M0R2), Proteus mirabilis (Q33DX5), Proteus mirabilis ARS68 (Q33DX5), Pseudomonas aeruginosa (A0MK31), Pseudomonas aeruginosa (Q8GRA1), Serratia marcescens (Q76G15), Streptoalloteichus hindustanus (O52472), Streptomyces kanamyceticus (Q75PS4)
brenda
Correa, L.L.; Witek, M.A.; Zelinskaya, N.; Picao, R.C.; Conn, G.L.
Heterologous expression and functional characterization of the exogenously acquired aminoglycoside resistance methyltransferases RmtD, RmtD2, and RmtG
Antimicrob. Agents Chemother.
60
699-702
2015
Escherichia coli, Micromonospora zionensis (Q7M0R2)
brenda