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Literature summary extracted from

  • 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 (2019), Antimicrob. Agents Chemother., 64, e01074-19 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.224 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 Clostridioides difficile

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.1.1.224 Mg2+ required Clostridioides difficile

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.224 2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin Clostridioides difficile
-
 S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
 ?
2.1.1.224 2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin Clostridioides difficile DF11
-
 S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.224 Clostridioides difficile
-
 clinical isolate from Mexico
-
2.1.1.224 Clostridioides difficile DF11
-
 clinical isolate from Mexico
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.224 recombinant tagged enzyme Cfr(E) from Escherichia coli strain Rosetta(DE3)pLysS by metal chelating chromatography Clostridioides difficile

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.224 2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
-
 Clostridioides difficile S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
 ?
2.1.1.224 2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin
-
 Clostridioides difficile DF11 S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
-
 ?
2.1.1.224 additional information in vitro assays with Escherichia coli RNA Clostridioides difficile ?
-
-
2.1.1.224 additional information in vitro assays with Escherichia coli RNA Clostridioides difficile DF11 ?
-
-

Synonyms

EC Number Synonyms Comment Organism
2.1.1.224 Cfr
-
 Clostridioides difficile
2.1.1.224 Cfr(E)
-
 Clostridioides difficile

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.224 37
-
 assay at Clostridioides difficile

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.224 8
-
 assay at Clostridioides difficile

Cofactor

EC Number Cofactor Comment Organism Structure
2.1.1.224 reduced [2Fe-2S] ferredoxin
-
 Clostridioides difficile
2.1.1.224 S-adenosyl-L-methionine
-
 Clostridioides difficile

General Information

EC Number General Information Comment Organism
2.1.1.224 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 Clostridioides difficile
2.1.1.224 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 Clostridioides difficile