Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
2 S-adenosyl-L-methionine + 2-methyladenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 2,8-dimethyladenine2503 in 23S rRNA
-
S-adenosylmethionine is both the methyl donor and the source of a 5'-deoxyadenosyl radical, which activates the substrate for methylation
incubation of Cfr with the wild-type 23S rRNA, already modified at the C2 position by the endogenous RlmN, provides 2,8-dimethyladenosine as the sole product
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
2 S-adenosyl-L-homocysteine + 2,8-dimethyladenine2503 in 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
S-adenosyl-L-methionine + adenine2503 in 23S rRNA + reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + oxidized [2Fe-2S] ferredoxin
-
-
-
?
additional information
?
-
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
verification of RNA methylation at A2503 in 23S rRNA by primer extension method
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
the enzyme methylates the 8 position of 23S rRNA residue A2503 to confer resistance to multiple antibiotic classes acting upon the large subunit of the bacterial ribosome
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
in the presence of sodium dithionite reconstituted Cfr is both reducible and able to cleave S-adenosyl-L-methionine to 5'-deoxyadeonsine, DOA
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
verification of RNA methylation at A2503 in 23S rRNA by primer extension method
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
verification of RNA methylation at A2503 in 23S rRNA by primer extension method
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
verification of RNA methylation at A2503 in 23S rRNA by primer extension method
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
verification of RNA methylation at A2503 in 23S rRNA by primer extension method
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
Cfr is an plasmid-acquired methyltransferase that protects cells from the action of antibiotics
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
S-adenosylmethionine is both the methyl donor and the source of a 5'-deoxyadenosyl radical, which activates the substrate for methylation. The enzyme can utilize protein-free 23S rRNA as a substrate, but not the fully assembled large ribosomal subunit, suggesting that the methylations take place during the assembly of the ribosome. The key recognition elements in the 23S rRNA are helices 90-92 and the adjacent single stranded RNA that encompasses A2503
identification of the reaction products as 8-methyladenine2503, and 2,8-dimethyladenine2503
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
the newly introduced methyl group is assembled from an S-adenosyl-L-methionine (SAM)-derived methylene fragment and a hydrogen atom that had migrated from the substrate amidine carbon. Rather than activating the adenosine nucleotide of the substrate by hydrogen atom abstraction from an amidine carbon, the 5'-deoxyadenosyl radical abstracts hydrogen from the second equivalent of SAM to form the SAM-derived radical cation. This species, or its corresponding sulfur ylide, subsequently adds into the substrate, initiating hydride shift and S-adenosylhomocysteine elimination to complete the formation of the methyl group
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
enzyme Cfr consumes two AdoMet equivalents per reaction cycle to support both methyl transfer to Cfr Cys338 (AdoMet1, step 1) and subsequently generation of the 59dA. radical (AdoMet2, step2)
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
two S-adenosyl-L-methionine molecules serve as the cofactor by providing the 5'-deoxyadenosyl radical for substrate activation and the methyl
-
-
?
additional information
?
-
kinetic analysis of methylation of a 155mer rRNA substrate by Cfr enzymes under limiting and under excess substrate conditions initiated with a protein-based reductant, substrate sequence, overview
-
-
-
additional information
?
-
kinetic analysis of methylation of a 155mer rRNA substrate by Cfr enzymes under limiting and under excess substrate conditions initiated with a protein-based reductant, substrate sequence, overview
-
-
-
additional information
?
-
-
in vitro assays with Escherichia coli RNA
-
-
-
additional information
?
-
kinetic analysis of methylation of a 155mer rRNA substrate by Cfr enzymes under limiting and under excess substrate conditions initiated with a protein-based reductant, substrate sequence, overview
-
-
-
additional information
?
-
-
in vitro assays with Escherichia coli RNA
-
-
-
additional information
?
-
kinetic analysis of methylation of a 155mer rRNA substrate by Cfr enzymes under limiting and under excess substrate conditions initiated with a protein-based reductant, substrate sequence, overview
-
-
-
additional information
?
-
kinetic analysis of methylation of a 155mer rRNA substrate by Cfr enzymes under limiting and under excess substrate conditions initiated with a protein-based reductant, substrate sequence, overview
-
-
-
additional information
?
-
kinetic analysis of methylation of a 155mer rRNA substrate by Cfr enzymes under limiting and under excess substrate conditions initiated with a protein-based reductant, substrate sequence, overview
-
-
-
additional information
?
-
Cfr confers a phenotype with resistance to phenicols, lincosamides, oxazolidinones, pleuromutilins, and streptogramin A antibiotics. Methylation of position 8 is the dominant antibiotic resistance determinant, but indigenous modification at position 2 also contributes to low-level resistance
-
-
?
additional information
?
-
Cfr methylates A2503 of 23S rRNA at C8 and C2, while RlmN only performs a C2 methylation using the same mechanism of function
-
-
-
additional information
?
-
kinetic analysis of methylation of a 155mer rRNA substrate by Cfr enzymes under limiting and under excess substrate conditions initiated with a protein-based reductant, substrate sequence, overview
-
-
-
additional information
?
-
kinetic analysis of methylation of a 155mer rRNA substrate by Cfr enzymes under limiting and under excess substrate conditions initiated with a protein-based reductant, substrate sequence, overview
-
-
-
additional information
?
-
-
enzyme Cfr methylates adenosine 2503 of the 23S rRNA in the peptidyltransferase centre (P-site) of the bacterial ribosome. In wild-type Cfr, where Cys338 is methylated, S-adenosyl-L-methionine binding leads to rapid oxidation of the [4Fe-4S] cluster and production of 5'-deoxyadenosine
-
-
?
additional information
?
-
methylation at C8 is similar to that at C2, cf. EC 2.1.1.192
-
-
?
additional information
?
-
the enzyme employs a [4Fe-4S] cluster to supply the requisite electron for reductive cleavage of SAM, usually to L-methionine and a 5'-deoxyadenosyl 5'-radical. SAM is the source of both the 5'-deoxyadenosyl 5'-radical and the appended methyl group
-
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
S-adenosyl-L-methionine + adenine2503 in 23S rRNA + reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + oxidized [2Fe-2S] ferredoxin
-
-
-
?
additional information
?
-
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
the enzyme methylates the 8 position of 23S rRNA residue A2503 to confer resistance to multiple antibiotic classes acting upon the large subunit of the bacterial ribosome
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
Cfr is an plasmid-acquired methyltransferase that protects cells from the action of antibiotics
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
-
?
additional information
?
-
Cfr confers a phenotype with resistance to phenicols, lincosamides, oxazolidinones, pleuromutilins, and streptogramin A antibiotics. Methylation of position 8 is the dominant antibiotic resistance determinant, but indigenous modification at position 2 also contributes to low-level resistance
-
-
?
additional information
?
-
-
enzyme Cfr methylates adenosine 2503 of the 23S rRNA in the peptidyltransferase centre (P-site) of the bacterial ribosome. In wild-type Cfr, where Cys338 is methylated, S-adenosyl-L-methionine binding leads to rapid oxidation of the [4Fe-4S] cluster and production of 5'-deoxyadenosine
-
-
?
additional information
?
-
methylation at C8 is similar to that at C2, cf. EC 2.1.1.192
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
evolution
-
bioinformatics analysis of the Cfr/RlmN family establishes their significant evolutionary link with radical-S-adenosyl-L-methionine enzymes. The RlmN subfamily is likely the ancestral form, whereas the Cfr subfamily arose via duplication and horizontal gene transfer
evolution
-
ClbA acts via the same mechanism as the Cfr methyltransferase
evolution
ClbB acts via the same mechanism as the Cfr methyltransferase
evolution
ClbC acts via the same mechanism as the Cfr methyltransferase
evolution
the cfr gene can be horizontally transferred to its hosts, as it is always found either on plasmids or together with insertion sequences. The cfr gene with only minor sequence differences are found worldwide in various bacteria isolated from humans and animals. Comparative sequence analysis identifies differentially conserved residues that indicate functional sequence divergence between the two classes of Cfr and RlmN-like sequences. The enzymes are homologous and use the same mechanism involving radical S-adenosyl methionine to methylate RNA via an intermediate involving a methylated cysteine in the enzyme and a transient cross-linking to the RNA, but they differ in which carbon atom in the adenine they methylate. The differentiation between the two classes is supported by experimental evidence from antibiotic resistance, primer extensions, and mass spectrometry. The Cfr- and RlmN-specific conserved sites provide a very good indication of whether a gene is Cfr-like or RlmN-like. Most bacteria have an rlmN-like gene and that all those that have a cfr-like gene also have an rlmN-like gene, evolutionary aspects of the bacterial distribution of Cfr and RlmN-like enzymes, overview
evolution
analysis of RNA methylation by phylogenetically diverse Cfr radical SAM enzymes reveals an ironbinding accessory domain in a clostridial enzyme. Sequence comparisons and phylogenetic analysis and tree, overview. Cfr homologues from Bacillus amyloliquefaciens, Enterococcus faecalis, Paenibacillus lautus, and Clostridioides difficile act as C8 adenine RNA methylases in biochemical assays
evolution
analysis of RNA methylation by phylogenetically diverse Cfr radical SAM enzymes reveals an ironbinding accessory domain in a clostridial enzyme. Sequence comparisons and phylogenetic analysis and tree, overview. Cfr homologues from Bacillus amyloliquefaciens, Enterococcus faecalis, Paenibacillus lautus, and Clostridioides difficile act as C8 adenine RNA methylases in biochemical assays
evolution
analysis of RNA methylation by phylogenetically diverse Cfr radical SAM enzymes reveals an ironbinding accessory domain in a clostridial enzyme. Sequence comparisons and phylogenetic analysis and tree, overview. Cfr homologues from Bacillus amyloliquefaciens, Enterococcus faecalis, Paenibacillus lautus, and Clostridioides difficile act as C8 adenine RNA methylases in biochemical assays
evolution
analysis of RNA methylation by phylogenetically diverse Cfr radical SAM enzymes reveals an ironbinding accessory domain in a clostridial enzyme. Sequence comparisons and phylogenetic analysis and tree, overview. Cfr homologues from Bacillus amyloliquefaciens, Enterococcus faecalis, Paenibacillus lautus, and Clostridioides difficile act as C8 adenine RNA methylases in biochemical assays. Clostridioides difficile Cfr contains an additional Cys-rich C-terminal domain that binds a mononuclear Fe2+ ion in a rubredoxin-type Cys4 motif, which has an important purpose for the observed C-terminal iron in the native fusion protein. Bioinformatic analysis of the Clostridioides difficile Cfr Cys-rich domain shows that it is widespread (about 1400 homologues) as a stand-alone gene in pathogenic or commensal Bacilli and Clostridia, with >10% encoded adjacent to a predicted radical SAM RNA methylase
evolution
-
cfr(C) is part of a putative 24 kb-transposon, which generated a detectable circular intermediate. An element differing by a single nucleotide is found in Clostridium difficile DA00203 from GenBank data, consistent with a recent horizontal transfer
evolution
enzyme Cfr belongs to the radical SAM (RS) superfamily of enzymes, catalysts that use S-adenosyl-L-methionine (SAM) as an oxidant to perform difficult and often complex transformations by radical mechanisms. RS superfamily enzymes employ a [4Fe-4S] cluster to supply the requisite electron for reductive cleavage of SAM, usually to L-methionine and a 5'-deoxyadenosyl 5'-radical. Similar enzyme RlmN (EC 2.1.1.192) is proposed to be an evolutionary precursor to Cfr. Residues conserved among both enzymes in a pairwise alignment of Escherichia coli RlmN and Staphylococcus aureus Cfr are mapped onto the RlmN structure. The catalytic residues in the active site are strictly conserved as are most of the surrounding residues within the core of the barrel, supporting the proposal that the enzymes use a common mechanism for C-methylation. The high degree of sequence conservation near the active site suggests that methylation site specificity during the reaction may be controlled in part by more distant structural elements. In Cfr, two large conformationally flexible regions in the RlmN structure are absent
evolution
evolutionary relationship between the Cfr and RlmN (EC 2.1.1.192) enzymes, phylogenetic analysis, overview
evolution
RlmN and Cfr belong to the radical SAM (RS) superfamily of enzymes. RlmN is proposed to be an evolutionary precursor to Cfr. The catalytic residues in theactive site are strictly conserved as are most of the surrounding residues within the core of the barrel
evolution
-
three cfr-like genes implicated in antibiotic resistance have been described, two of which, cfr(B) and cfr(C), have been sporadically detected in Clostridium difficile. The methylase activity of Cfr(C) has not been confirmed. cfr(B), cfr(C), and a cfr-like genes show only 51 to 58% protein sequence identity to Cfr and Cfr-like enzymes in clinical Clostridium difficile isolates recovered across nearly a decade in Mexico, Honduras, Costa Rica, and Chile. This resistance gene is termed cfr(E). The predicted protein sequence of Cfr(E) shows homology to C8 RNA-methylating enzymes. Enzymes Cfr(C) or Cfr(E) are determined to methylate A2503 at the C8 position. The cfr-like gene of isolate DF11 (Cfr(E))is found integrated into an undescribed MGE that shows partial hits to genomic sequences of various intestinal Firmicutes, but in all cases, shared regions do not include cfr(E) or its adjacent genes, overview
evolution
-
analysis of RNA methylation by phylogenetically diverse Cfr radical SAM enzymes reveals an ironbinding accessory domain in a clostridial enzyme. Sequence comparisons and phylogenetic analysis and tree, overview. Cfr homologues from Bacillus amyloliquefaciens, Enterococcus faecalis, Paenibacillus lautus, and Clostridioides difficile act as C8 adenine RNA methylases in biochemical assays
-
evolution
-
analysis of RNA methylation by phylogenetically diverse Cfr radical SAM enzymes reveals an ironbinding accessory domain in a clostridial enzyme. Sequence comparisons and phylogenetic analysis and tree, overview. Cfr homologues from Bacillus amyloliquefaciens, Enterococcus faecalis, Paenibacillus lautus, and Clostridioides difficile act as C8 adenine RNA methylases in biochemical assays
-
evolution
-
cfr(C) is part of a putative 24 kb-transposon, which generated a detectable circular intermediate. An element differing by a single nucleotide is found in Clostridium difficile DA00203 from GenBank data, consistent with a recent horizontal transfer
-
evolution
-
analysis of RNA methylation by phylogenetically diverse Cfr radical SAM enzymes reveals an ironbinding accessory domain in a clostridial enzyme. Sequence comparisons and phylogenetic analysis and tree, overview. Cfr homologues from Bacillus amyloliquefaciens, Enterococcus faecalis, Paenibacillus lautus, and Clostridioides difficile act as C8 adenine RNA methylases in biochemical assays
-
evolution
-
ClbA acts via the same mechanism as the Cfr methyltransferase
-
evolution
-
ClbB acts via the same mechanism as the Cfr methyltransferase
-
evolution
-
analysis of RNA methylation by phylogenetically diverse Cfr radical SAM enzymes reveals an ironbinding accessory domain in a clostridial enzyme. Sequence comparisons and phylogenetic analysis and tree, overview. Cfr homologues from Bacillus amyloliquefaciens, Enterococcus faecalis, Paenibacillus lautus, and Clostridioides difficile act as C8 adenine RNA methylases in biochemical assays. Clostridioides difficile Cfr contains an additional Cys-rich C-terminal domain that binds a mononuclear Fe2+ ion in a rubredoxin-type Cys4 motif, which has an important purpose for the observed C-terminal iron in the native fusion protein. Bioinformatic analysis of the Clostridioides difficile Cfr Cys-rich domain shows that it is widespread (about 1400 homologues) as a stand-alone gene in pathogenic or commensal Bacilli and Clostridia, with >10% encoded adjacent to a predicted radical SAM RNA methylase
-
evolution
-
three cfr-like genes implicated in antibiotic resistance have been described, two of which, cfr(B) and cfr(C), have been sporadically detected in Clostridium difficile. The methylase activity of Cfr(C) has not been confirmed. cfr(B), cfr(C), and a cfr-like genes show only 51 to 58% protein sequence identity to Cfr and Cfr-like enzymes in clinical Clostridium difficile isolates recovered across nearly a decade in Mexico, Honduras, Costa Rica, and Chile. This resistance gene is termed cfr(E). The predicted protein sequence of Cfr(E) shows homology to C8 RNA-methylating enzymes. Enzymes Cfr(C) or Cfr(E) are determined to methylate A2503 at the C8 position. The cfr-like gene of isolate DF11 (Cfr(E))is found integrated into an undescribed MGE that shows partial hits to genomic sequences of various intestinal Firmicutes, but in all cases, shared regions do not include cfr(E) or its adjacent genes, overview
-
malfunction
Clostridioides difficile Cfr contains an additional Cys-rich C-terminal domain that binds a mononuclear Fe2+ ion in a rubredoxin-type Cys4 motif. The C-terminal domain can be truncated with minimal impact on Cfr activity, but the rate of turnover is decreased upon disruption of the Fe2+-binding site by Zn2+ substitution or ligand mutation
malfunction
-
Clostridioides difficile Cfr contains an additional Cys-rich C-terminal domain that binds a mononuclear Fe2+ ion in a rubredoxin-type Cys4 motif. The C-terminal domain can be truncated with minimal impact on Cfr activity, but the rate of turnover is decreased upon disruption of the Fe2+-binding site by Zn2+ substitution or ligand mutation
-
metabolism
methyl transfer is essential in the synthesis of cellular metabolites and clinically relevant natural products, and in the modification of RNA, DNA, lipids, and proteins
metabolism
-
several groups of antibiotics inhibit bacterial growth by binding to bacterial ribosomes. Mutations in ribosomal protein L3 have been associated with resistance to linezolid and tiamulin, which both bind at the peptidyl transferase center in the ribosome. Resistance to these and other antibiotics also occurs through methylation of 23S rRNA at position A2503 by the methyltransferase Cfr. The resistance from Cfr is, in all cases, stronger than the effects of the L3 mutations, but various effects are obtained with the combinations of Cfr and L3 mutations ranging from a synergistic to an antagonistic effect. Linezolid and tiamulin susceptibility vary greatly among the L3 mutations, while no significant effects on florfenicol and Q-D susceptibility are seen. Relative positions of L3 mutations, methylation of the 23S rRNA at position A2503, and antibiotics, three-dimensional structure model, overview. Analysis of antibiotic susceptibilities of the L3 mutant strains with and without Cfr expression
physiological function
Cfr confers a phenotype with resistance to phenicols, lincosamides, oxazolidinones, pleuromutilins, and streptogramin A antibiotics. The RlmN knock out strain JW2501-1 is less sensitive to the antibiotics than standard laboratory strain HB101
physiological function
-
cfr confers combined resistance to chloramphenicol, florfenicol and clindamycin
physiological function
-
the Cfr rRNA methyltransferase confers resistance to phenicols, lincosamides, oxazolidinones, pleuromutilins, and streptogramin A antibiotics
physiological function
-
ClbA confers resistance to antibiotics, florfenicol, clindamycin, linezolid, tiamulin, and streptogramin A/streptogramin B, to the cell, also when expressed in Escherichia coli strain AS19, overview
physiological function
ClbB confers resistance to antibiotics, florfenicol, clindamycin, linezolid, tiamulin, and streptogramin A/streptogramin B, to the cell, also when expressed in Escherichia coli strain AS19, overview
physiological function
ClbC confers resistance to antibiotics, florfenicol, clindamycin, linezolid, tiamulin, and streptogramin A/streptogramin B, to the cell, also when expressed in Escherichia coli strain AS19, overview
physiological function
-
rRNA methyltransferase Cfr that methylates the conserved 23S rRNA residue A2503, located in a functionally critical region of the ribosome, confers resistance to an array of ribosomal antibiotics, including linezolid
physiological function
-
the enzyme methylates the 8 position of 23S rRNA residue A2503 to confer resistance to multiple antibiotic classes acting upon the large subunit of the bacterial ribosome. Radical-SAM enzymes use an Fe-S cluster to generate the 5'-deoxyadenosyl radical from SAM, enabling them to modify intrinsically unreactive centres such as adenosine C8
physiological function
-
enzyme Cfr methylates adenosine 2503 of the 23S rRNA in the peptidyltransferase centre (P-site) of the bacterial ribosome. This modification protects host bacteria, notably methicillin-resistant Staphylococcus aureus (MRSA), from numerous antibiotics, including agents (e.g. linezolid, retapamulin)
physiological function
the Cfr methyltransferase primarily methylates C-8 in A2503 of 23S rRNA in the peptidyl transferase region of bacterial ribosomes. Enzyme Cfr confers resistance to antibiotics binding to the peptidyl transferase center on the ribosome, defining a PhLOPSa phenotype that reflects resistance to phenicol, lincosamide, oxazolidinone, pleuromutilin, and streptogramin A antibiotic classes. Cfr also provides resistance to some large macrolide antibiotics. The cfr gene is thus a health threat when spreading in pathogenic bacteria because many clinically important antibiotics become useless for treatment
physiological function
the Cfr RNA methyltransferase causes multiple resistances to peptidyl transferase inhibitors by methylation of A2503 23S rRNA
physiological function
antibiotic resistance effects of wild-type and mutant enzymes, overview
physiological function
-
Cfr is a radical S-adenosyl-L-methionine (SAM) enzyme that confers cross-resistance to antibiotics targeting the 23S rRNA through hypermethylation of nucleotide A2503
physiological function
Cfr is a radical S-adenosylmethionine (SAM) RNA methylase linked to multidrug antibiotic resistance in bacterial pathogens. It catalyzes a chemically challenging C-C bond-forming reaction to methylate C8 of A2503 (Escherichia coli numbering) of 23S rRNA during ribosome assembly
physiological function
Cfr is a radical S-adenosylmethionine (SAM) RNA methylase linked to multidrug antibiotic resistance in bacterial pathogens. It catalyzes a chemically challenging C-C bond-forming reaction to methylate C8 of A2503 (Escherichia coli numbering) of 23S rRNA during ribosome assembly
physiological function
Cfr is a radical S-adenosylmethionine (SAM) RNA methylase linked to multidrug antibiotic resistance in bacterial pathogens. It catalyzes a chemically challenging C-C bond-forming reaction to methylate C8 of A2503 (Escherichia coli numbering) of 23S rRNA during ribosome assembly
physiological function
Cfr is a radical S-adenosylmethionine (SAM) RNA methylase linked to multidrug antibiotic resistance in bacterial pathogens. It catalyzes a chemically challenging C-C bond-forming reaction to methylate C8 of A2503 (Escherichia coli numbering) of 23S rRNA during ribosome assembly
physiological function
-
Cfr modifies C8 of A2503 in 23S rRNA conferring resistance to multiple classes of antibiotics. A2503 is also methylated at C8 by RlmN, which is both evolutionarily and mechanistically related to Cfr. Methylation of C8 of A2503 is the only known in vivo activity of Cfr, while RlmN also installs a C2 methyl group at adenosine 37
physiological function
-
Clostridium bolteae strain 90B3 is multiresistant to various antibiotics including ampicillin, as well as to linezolid, florfenicol, streptogramin A, and tiamulin due to methylation of 23S rRNA at A2503
physiological function
gene cfr(C) confers linezolid resistance is common in Clostridium difficile. Clostridium difficile strain T10 is resistant to erythromycin, chloramphenicol, clindamycin, florfenicol, linezolid, streptogramin A, and tiamulin due to methylation of 23S rRNA at A2503
physiological function
-
the cfr gene encodes an rRNA methyltransferase that adds a methyl group at the C-8 position of 23S rRNA nucleotide A2503 at the peptidyl transferase center (PTC) in the ribosome. This m8A2503 modification confers resistance to more than six classes of antibiotics that bind at overlapping nonidentical sites at the PTC
physiological function
the radical SAM (RS) enzymes RlmN and Cfr methylate 23S ribosomal RNA, modifying the C2 or C8 position of adenosine 2503. The methyl groups are installed by a two-step sequence involving initial methylation of a conserved Cys residue (RlmN Cys 355) by SAM. Methyl transfer to the substrate requires reductive cleavage of a second equivalent of SAM. Cfr confers antibiotic resistance by methylating C8 of A2503
physiological function
-
Cfr is a radical S-adenosylmethionine (SAM) RNA methylase linked to multidrug antibiotic resistance in bacterial pathogens. It catalyzes a chemically challenging C-C bond-forming reaction to methylate C8 of A2503 (Escherichia coli numbering) of 23S rRNA during ribosome assembly
-
physiological function
-
gene cfr(C) confers linezolid resistance is common in Clostridium difficile. Clostridium difficile strain T10 is resistant to erythromycin, chloramphenicol, clindamycin, florfenicol, linezolid, streptogramin A, and tiamulin due to methylation of 23S rRNA at A2503
-
physiological function
-
Cfr is a radical S-adenosylmethionine (SAM) RNA methylase linked to multidrug antibiotic resistance in bacterial pathogens. It catalyzes a chemically challenging C-C bond-forming reaction to methylate C8 of A2503 (Escherichia coli numbering) of 23S rRNA during ribosome assembly
-
physiological function
-
Clostridium bolteae strain 90B3 is multiresistant to various antibiotics including ampicillin, as well as to linezolid, florfenicol, streptogramin A, and tiamulin due to methylation of 23S rRNA at A2503
-
physiological function
-
Cfr is a radical S-adenosylmethionine (SAM) RNA methylase linked to multidrug antibiotic resistance in bacterial pathogens. It catalyzes a chemically challenging C-C bond-forming reaction to methylate C8 of A2503 (Escherichia coli numbering) of 23S rRNA during ribosome assembly
-
physiological function
-
ClbA confers resistance to antibiotics, florfenicol, clindamycin, linezolid, tiamulin, and streptogramin A/streptogramin B, to the cell, also when expressed in Escherichia coli strain AS19, overview
-
physiological function
-
ClbB confers resistance to antibiotics, florfenicol, clindamycin, linezolid, tiamulin, and streptogramin A/streptogramin B, to the cell, also when expressed in Escherichia coli strain AS19, overview
-
physiological function
-
Cfr is a radical S-adenosylmethionine (SAM) RNA methylase linked to multidrug antibiotic resistance in bacterial pathogens. It catalyzes a chemically challenging C-C bond-forming reaction to methylate C8 of A2503 (Escherichia coli numbering) of 23S rRNA during ribosome assembly
-
physiological function
-
Cfr is a radical S-adenosyl-L-methionine (SAM) enzyme that confers cross-resistance to antibiotics targeting the 23S rRNA through hypermethylation of nucleotide A2503
-
physiological function
-
rRNA methyltransferase Cfr that methylates the conserved 23S rRNA residue A2503, located in a functionally critical region of the ribosome, confers resistance to an array of ribosomal antibiotics, including linezolid
-
additional information
-
acquisition of the cfr gene does not produce any appreciable reduction in the cell growth rate, analysis of fitness cost of cfr expression, overview. Genes ermB and cfr are coexpressed under the Perm promoter in the mlr operon. Dimethylation of A2058 by the Erm methyltransferase increases the fitness cost associated with Cfr-mediated modification of A2503
additional information
-
the presence of a methyl group on Cfr Cys338 is a key determinant of the activity of the enzyme towards S-adenosyl-L-methionine, thus enabling a single active site to support two distinct modes of S-adenosyl-L-methionine cleavage
additional information
-
although SAM is the source of the appended methyl carbon in the reactions catalyzed by RlmN and Cfr, these enzymes operate by a mechanism that is distinctly different from that of typical SAM-dependent methyltransferases. As radical SAM (RS) enzymes, RlmN and Cfr employ very similar radical-based mechanisms of catalysis, initiated by the abstraction of a hydrogen atom from a Cys-appended methyl group via a 5'-deoxyadenosyl 5'-radical. Subsequent attack of the resulting methylene radical upon the carbon atom undergoing methylation affords a protein/RNA cross-linked intermediate whose resolution requires prior proton abstraction from C2 (RlmN) or C8 (Cfr) of the substrate by an unidentified base. Conversion of the intermediate to the methylated product has also been demonstrated in the Cfr reaction. The proximity (5.0 A) of the Cys 355 side chain (the proposed site of thiyl radical formation) to the sulfur atom of Met176, a strictly conserved residue in RlmN and Cfr, might allow formation of a transient thiosulfuranyl radical
additional information
gene cfr(C) is mainly confined in Clostridium difficile within polymorphic environments suggesting that this microorganism is a reservoir for PhLOPSA resistance. In silico analysis shows cfr(C) in 19 out of 274 Clostridium difficile genomes. This gene is also detected by PCR analysis in 9 out of 80 Clostridium difficile from a laboratory strain collection according to resistance to linezolid and florfenicol
additional information
-
gene cfr(C) is mainly confined in Clostridium difficile within polymorphic environments suggesting that this microorganism is a reservoir for PhLOPSA resistance. In silico analysis shows cfr(C) in 19 out of 274 Clostridium difficile genomes. This gene is also detected by PCR analysis in 9 out of 80 Clostridium difficile from a laboratory strain collection according to resistance to linezolid and florfenicol
additional information
mechanisms of catalytic action of Cfr and related RlmN (EC 2.1.1.192), the methylation mechanism involves a transitory methylation of Cys338 for Cfr and Cys355 for RlmN, investigation of target binding to the active sites of the two enzymes, overview. Cfr and RlmN are methylated before transfer of the methyl group to the substrate. Homology structure modelling, molecular dynamics simulations, and calculation of the binding free energy, using structure of Escherichia coli RlmN (PDB ID 3RFA), the homology model is made with the [4Fe-4S] cluster and a SAM molecule positioned in the same way as seen in the RlmN X-ray structure. Defining regions of the active site to be interchanged to investigate C8/C2 specificity
additional information
structure-function analysis of RlmN from Escherichia coli (EC 2.1.1.192) compared to Cfr, Escherichia coli RlmN and Staphylococcus aureus Cfr are mapped onto the RlmN structure, detailed overview. The Cfr reaction proceeds by a ping-pong mechanism. The methyl group from one SAM molecule is initially appended to a conserved Cys residue by a typical SN2 displacement. This SAM-derived one carbon unit is then attached to the RNA by radical addition initiated by a 5'-deoxyadenosyl 5'-radical formed from a second molecule of SAM. The expected role of the radical is to abstract a hydrogen atom from the substrate, in this case the C8 (Cfr) hydrogen atom from A2503, activating the substrate for subsequent methylation. Finally, this covalent intermediate is resolved by formation of a disulfide bond between the methyl-carrying Cys (mCys) residue and a second conserved Cys residue. Cys 355 is a key catalytic residue that is methylated in the first step of the proposed mechanism
additional information
-
gene cfr(C) is mainly confined in Clostridium difficile within polymorphic environments suggesting that this microorganism is a reservoir for PhLOPSA resistance. In silico analysis shows cfr(C) in 19 out of 274 Clostridium difficile genomes. This gene is also detected by PCR analysis in 9 out of 80 Clostridium difficile from a laboratory strain collection according to resistance to linezolid and florfenicol
-
additional information
-
acquisition of the cfr gene does not produce any appreciable reduction in the cell growth rate, analysis of fitness cost of cfr expression, overview. Genes ermB and cfr are coexpressed under the Perm promoter in the mlr operon. Dimethylation of A2058 by the Erm methyltransferase increases the fitness cost associated with Cfr-mediated modification of A2503
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
a plasmid-borne cfr geneis transfromed into a uL3-depleted Escherichia coli strain containing either wild-type L3 or L3 with one of seven mutations, G147R, Q148F, N149S, N149D, N149R, Q150L, or T151P, expressed from plasmid-carried rplC genes. The L3 mutations are well tolerated, with small to moderate growth rate decreases. The presence of Cfr has a very minor influence on the growth rate. The transformants show resistance to linezolid, tiamulin, florfenicol, and Synercid (a combination of quinupristin and dalfopristin [Q-D])
-
expressed as C-terminally His6-tagged enzyme, wild-type and mutant enzymes C125A, C129A and C132A
-
expressed in Escherichia coli Rosetta2(DE3)pLysS cells
gene cfr(C), DNA and amino acid sequence determination and analysis, quantitative RT-PCR expression analysis, cfr(C) is constitutively transcribed, gene cfr(C) is part of a putative 24 kb-transposon, which generates a detectable circular intermediate
-
gene cfr(C), DNA and amino acid sequence determination and analysis, the cfr-like is cloned in Clostridium difficile strain 630DELTAerm in which it confers resistance to phenicols, lincosamides, oxazolidinones, pleuromutilins, and streptogramin A (PhLOPSA)
gene cfr(E), DNA and amino acid sequence determination and analysis, genotyping, sequence comparisons and phylogenetic analysis of different cfr gene variants, recombinant expression of tagged enzyme Cfr(E) in Escherichia coli strain Rosetta(DE3)pLysS
-
gene cfr, operon isc, co-expression with isc proteins, expression of His6-tagged Cfr under control of an arabinose-inducible promoter in Escherichia coli strain BL21Star
-
gene cfr, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain +Rosetta2(DE3)
-
gene cfr, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain JW2501-1 (rlmN knockout) and AS19
gene cfr, sequence comparisons and phylogenetic analysis and tree, overview
gene cfr, sequence comparisons and phylogenetic analysis, primer extension analysis, cloning and expression in Escherichia coli strains TOP10 and AS19
gene cfr, The Cfr-like protein in Peptoclostridium difficile is hosted by a transposon, and the same gene is found in other strains, sequence comparisons, overview. Recombinant expression in Escherichia coli strain AS19 which then shows elevated MIC values for five classes of antibiotics, and recombinant expression in rlmN-deficient Escherichia coli strain JW2505-1
gene cfr, while cfr is typically plasmid borne, in CM05 it is located on the chromosome and is coexpressed with ermB as part of the mlr operon. DNA and amino acid sequence determination and analysis and genetic organization, overview. The cfr-containing mlr operon is located within a 15.5-kb plasmid-like insertion into 23S rRNA allele 4. In the LZDs colonies, designated CM05DELTA, a recombination event involving the two ermB genes had occurred, resulting in the deletion of cfr and the 3= flanking region, cfr-istAS-istBS-ermB
gene clbA, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression in Escherichia coli strain AS19 using plasmid pLJ10
-
gene clbB, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression in Escherichia coli strain AS19 using plasmid pLJ10
gene clbC, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression in Escherichia coli strain AS19 using plasmid pLJ10
gene gfr, ermB and cfr genes are naturally coexpressed under the Perm promoter in the mlr operon, expression of mlr results in modification of A2058 and A2503 in 23S rRNA and renders cells resistant to all clinically relevant antibiotics that target the large ribosomal subunit
-
gene cfr, sequence comparisons and phylogenetic analysis and tree, overview
gene cfr, sequence comparisons and phylogenetic analysis and tree, overview
gene cfr, sequence comparisons and phylogenetic analysis and tree, overview
gene cfr, sequence comparisons and phylogenetic analysis and tree, overview
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Long, K.S.; Poehlsgaard, J.; Kehrenberg, C.; Schwarz, S.; Vester, B.
The Cfr rRNA methyltransferase confers resistance to phenicols, lincosamides, oxazolidinones, pleuromutilins, and streptogramin A antibiotics
Antimicrob. Agents Chemother.
50
2500-2505
2006
Escherichia coli
brenda
Yan, F.; LaMarre, J.M.; Rhrich, R.; Wiesner, J.; Jomaa, H.; Mankin, A.S.; Fujimori, D.G.
RlmN and Cfr are radical SAM enzymes involved in methylation of ribosomal RNA
J. Am. Chem. Soc.
132
3953-3964
2010
Escherichia coli
brenda
Kehrenberg, C.; Schwarz, S.; Jacobsen, L.; Hansen, L.H.; Vester, B.
A new mechanism for chloramphenicol, florfenicol and clindamycin resistance: methylation of 23S ribosomal RNA at A2503
Mol. Microbiol.
57
1064-1073
2005
Escherichia coli
brenda
Kaminska, K.H.; Purta, E.; Hansen, L.H.; Bujnicki, J.M.; Vester, B.; Long, K.S.
Insights into the structure, function and evolution of the radical-SAM 23S rRNA methyltransferase Cfr that confers antibiotic resistance in bacteria
Nucleic Acids Res.
38
1652-1663
2009
Escherichia coli
brenda
Giessing, A.M.; Jensen, S.S.; Rasmussen, A.; Hansen, L.H.; Gondela, A.; Long, K.; Vester, B.; Kirpekar, F.
Identification of 8-methyladenosine as the modification catalyzed by the radical SAM methyltransferase Cfr that confers antibiotic resistance in bacteria
RNA
15
327-336
2009
Escherichia coli K-12 (P36979)
brenda
Yan, F.; Fujimori, D.G.
RNA methylation by radical SAM enzymes RlmN and Cfr proceeds via methylene transfer and hydride shift
Proc. Natl. Acad. Sci. USA
108
3930-3934
2011
Escherichia coli
brenda
Boal, A.K.; Grove, T.L.; McLaughlin, M.I.; Yennawar, N.H.; Booker, S.J.; Rosenzweig, A.C.
Structural basis for methyl transfer by a radical SAM enzyme
Science
332
1089-1092
2011
Escherichia coli
brenda
Grove, T.L.; Benner, J.S.; Radle, M.I.; Ahlum, J.H.; Landgraf, B.J.; Krebs, C.; Booker, S.J.
A radically different mechanism for S-adenosylmethionine-dependent methyltransferases
Science
332
604-607
2011
Escherichia coli
brenda
LaMarre, J.M.; Locke, J.B.; Shaw, K.J.; Mankin, A.S.
Low fitness cost of the multidrug resistance gene cfr
Antimicrob. Agents Chemother.
55
3714-3719
2011
Staphylococcus aureus, Staphylococcus aureus RN4220
brenda
Locke, J.B.; Rahawi, S.; Lamarre, J.; Mankin, A.S.; Shaw, K.J.
Genetic environment and stability of cfr in methicillin-resistant Staphylococcus aureus CM05
Antimicrob. Agents Chemother.
56
332-340
2012
Staphylococcus aureus (E1VCY5), Staphylococcus aureus, Staphylococcus aureus CM05 (E1VCY5), Staphylococcus aureus CM05
brenda
Hansen, L.H.; Planellas, M.H.; Long, K.S.; Vester, B.
The order Bacillales hosts functional homologs of the worrisome cfr antibiotic resistance gene
Antimicrob. Agents Chemother.
56
3563-3567
2012
Bacillus amyloliquefaciens, Brevibacillus brevis (C0ZJA6), Alkalihalobacillus clausii (Q5WJ42), Bacillus amyloliquefaciens FZB42, Brevibacillus brevis NBRC 100599 (C0ZJA6)
brenda
Booth, M.P.; Challand, M.R.; Emery, D.C.; Roach, P.L.; Spencer, J.
High-level expression and reconstitution of active Cfr, a radical-SAM rRNA methyltransferase that confers resistance to ribosome-acting antibiotics
Protein Expr. Purif.
74
204-210
2010
Azotobacter vinelandii
brenda
Atkinson, G.C.; Hansen, L.H.; Tenson, T.; Rasmussen, A.; Kirpekar, F.; Vester, B.
Distinction between the Cfr methyltransferase conferring antibiotic resistance and the housekeeping RlmN methyltransferase
Antimicrob. Agents Chemother.
57
4019-4026
2013
Mammaliicoccus sciuri (Q9FBG4)
brenda
Challand, M.R.; Salvadori, E.; Driesener, R.C.; Kay, C.W.; Roach, P.L.; Spencer, J.
Cysteine methylation controls radical generation in the Cfr radical AdoMet rRNA methyltransferase
PLoS ONE
8
e67979
2013
Staphylococcus aureus, Mammaliicoccus sciuri (Q9FBG4)
brenda
Hansen, L.H.; Vester, B.
A cfr-like gene from Clostridium difficile confers multiple antibiotic resistance by the same mechanism as the cfr gene
Antimicrob. Agents Chemother.
59
5841-5843
2015
Clostridioides difficile (V5ZEZ1), Clostridioides difficile
brenda
Wang, C.; Yang, X.; Wang, E.; Li, B.
Quantum chemistry studies of adenosine 2503 methylation by S-adenosylmethionine-dependent enzymes
Int. J. Quantum Chem.
113
1409-1415
2013
Staphylococcus aureus (A5HBL2)
-
brenda
Pakula, K.K.; Hansen, L.H.; Vester, B.
Combined effect of the Cfr methyltransferase and ribosomal protein L3 mutations on resistance to ribosome-targeting antibiotics
Antimicrob. Agents Chemother.
61
e00862-17
2017
Staphylococcus sp.
brenda
Stojkovic, V.; Ulate, M.F.; Hidalgo-Villeda, F.; Aguilar, E.; Monge-Cascante, C.; Pizarro-Guajardo, M.; Tsai, K.; Tzoc, E.; Camorlinga, M.; Paredes-Sabja, D.; Quesada-Gomez, C.; Fujimori, D.G.; Rodriguez, C.
cfr(B), cfr(C), and a new cfr-like gene, cfr(E), in Clostridium difficile strains recovered across Latin America
Antimicrob. Agents Chemother.
64
e01074-19
2019
Clostridioides difficile, Clostridioides difficile DF11
brenda
Candela, T.; Marvaud, J.; Nguyen, T.; Lambert, T.
A cfr-like gene cfr(C) conferring linezolid resistance is common in Clostridium difficile
Int. J. Antimicrob. Agents
50
496-500
2017
Enterocloster bolteae, Clostridioides difficile (V5ZEQ2), Clostridioides difficile, Clostridioides difficile T10 (V5ZEQ2), Clostridioides difficile T10, Enterocloster bolteae 90B3
brenda
Ntokou, E.; Hansen, L.; Kongsted, J.; Vester, B.
Biochemical and computational analysis of the substrate specificities of Cfr and RlmN Methyltransferases
PLoS ONE
10
e0145655
2015
Mammaliicoccus sciuri (Q9FBG4)
brenda
Fitzsimmons, C.M.; Fujimori, D.G.
Determinants of tRNA recognition by the radical SAM enzyme RlmN
PLoS ONE
11
e0167298
2016
Enterococcus faecalis (A0A2Z6BLR4), Bacillus amyloliquefaciens (A0A4V7TJX0), Paenibacillus lautus (D3EIG6), Clostridioides difficile (V5ZEQ2), Bacillus amyloliquefaciens DSM-7 (A0A4V7TJX0), Paenibacillus lautus Y412MC10 (D3EIG6), Enterococcus faecalis TX0645 (A0A2Z6BLR4), Clostridioides difficile QCD63q42 (V5ZEQ2)
brenda
Boal, A.K.; Grove, T.L.; McLaughlin, M.I.; Yennawar, N.H.; Booker, S.J.; Rosenzweig, A.C.
Structural basis for methyl transfer by a radical SAM enzyme
Science
332
1089-1092
2011
Staphylococcus aureus (A5HBL2)
brenda
Schwalm, E.; Grove, T.; Booker, S.; Boal, A.
Crystallographic capture of a radical S-adenosylmethionine enzyme in the act of modifying tRNA
Science
352
309-312
2016
Escherichia coli
brenda