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Literature summary for 2.1.1.202 extracted from
- Multisite-specific tRNA:m5C-methyltransferase (Trm4) in yeast Saccharomyces cerevisiae: identification of the gene and substrate specificity of the enzyme (1999), RNA, 5, 1105-1118.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| inserted in the expression vector pET28b, and the corresponding protein is hyperexpressed in Escherichia coli BL21(DE3) | Saccharomyces cerevisiae |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 77879 | - |
x * 77879, calculated from sequence | Saccharomyces cerevisiae |
| 80200 | - |
x * 80200, calculated from sequence, including Hi26- and thrombin cleavage sequences | Saccharomyces cerevisiae |
| 82200 | - |
x * 82200, SDS-PAGE, recombinant enzyme including Hi26- and thrombin cleavage sequences | Saccharomyces cerevisiae |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| S-adenosyl-L-methionine + cytosine34 in tRNA | Saccharomyces cerevisiae | the enzyme is responsible for complete m5C methylation of yeast tRNA. The frequency of modification depends on the cytosine position in tRNA. At positions 34 and 40, m5C is found only in two yeast tRNAs (tRNALeu (CUA) and tRNAPhe (GAA), respectively), whereas most other elongator yeast tRNAs bear either m5C48 or m5C49, but never both in the same tRNA molecule | S-adenosyl-L-homocysteine + 5-methylcytosine34 in tRNA | - |
? |  |
| S-adenosyl-L-methionine + cytosine40 in tRNA | Saccharomyces cerevisiae | the enzyme is responsible for complete m5C methylation of yeast tRNA. The frequency of modification depends on the cytosine position in tRNA. At positions 34 and 40, m5C is found only in two yeast tRNAs (tRNALeu (CUA) and tRNAPhe (GAA), respectively), whereas most other elongator yeast tRNAs bear either m5C48 or m5C49, but never both in the same tRNA molecule | S-adenosyl-L-homocysteine + 5-methylcytosine40 in tRNA | - |
? | |
| S-adenosyl-L-methionine + cytosine48 in tRNA | Saccharomyces cerevisiae | the enzyme is responsible for complete m5C methylation of yeast tRNA. The frequency of modification depends on the cytosine position in tRNA. At positions 34 and 40, m5C is found only in two yeast tRNAs (tRNALeu (CUA) and tRNAPhe (GAA), respectively), whereas most other elongator yeast tRNAs bear either m5C48 or m5C49, but never both in the same tRNA molecule | S-adenosyl-L-homocysteine + 5-methylcytosine48 in tRNA | - |
? | |
| S-adenosyl-L-methionine + cytosine49 in tRNA | Saccharomyces cerevisiae | the enzyme is responsible for complete m5C methylation of yeast tRNA. The frequency of modification depends on the cytosine position in tRNA. At positions 34 and 40, m5C is found only in two yeast tRNAs (tRNALeu (CUA) and tRNAPhe (GAA), respectively), whereas most other elongator yeast tRNAs bear either m5C48 or m5C49, but never both in the same tRNA molecule | S-adenosyl-L-homocysteine + 5-methylcytosine49 in tRNA | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Saccharomyces cerevisiae | P38205 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant Ybl024p is purified to apparent homogeneity by one-step affinity chromatography on Ni21-NTA-agarose column | Saccharomyces cerevisiae |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| S-adenosyl-L-methionine + cytosine34 in tRNA precursor = S-adenosyl-L-homocysteine + 5-methylcytosine34 in tRNA precursor | the formation of 5-methylcytosine34 is a strictly intron-dependent process | Saccharomyces cerevisiae | |
| S-adenosyl-L-methionine + cytosine40 in tRNA precursor = S-adenosyl-L-homocysteine + 5-methylcytosine40 in tRNA precursor | the formation of 5-methylcytosine34 is a strictly intron-dependent process | Saccharomyces cerevisiae | |
| S-adenosyl-L-methionine + cytosine48 in tRNA = S-adenosyl-L-homocysteine + 5-methylcytosine48 in tRNA | the formation of 5-methylcytosine48 is an intron-independent process | Saccharomyces cerevisiae | |
| S-adenosyl-L-methionine + cytosine49 in tRNA = S-adenosyl-L-homocysteine + 5-methylcytosine49 in tRNA | the formation of 5-methylcytosine49 is an intron-independent process | Saccharomyces cerevisiae |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| S-adenosyl-L-methionine + cytosine34 in tRNA | the enzyme is responsible for complete m5C methylation of yeast tRNA. The frequency of modification depends on the cytosine position in tRNA. At positions 34 and 40, m5C is found only in two yeast tRNAs (tRNALeu (CUA) and tRNAPhe (GAA), respectively), whereas most other elongator yeast tRNAs bear either m5C48 or m5C49, but never both in the same tRNA molecule | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + 5-methylcytosine34 in tRNA | - |
? | |
| S-adenosyl-L-methionine + cytosine34 in tRNA precursor | mini-tRNALeu (composed of the anticodon stem-loop extended by the intron) is used to test the formation of m5C34. The formation of m5C34 is strictly intron dependent | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + 5-methylcytosine34 in tRNA precursor | - |
? | |
| S-adenosyl-L-methionine + cytosine40 in tRNA | the enzyme is responsible for complete m5C methylation of yeast tRNA. The frequency of modification depends on the cytosine position in tRNA. At positions 34 and 40, m5C is found only in two yeast tRNAs (tRNALeu (CUA) and tRNAPhe (GAA), respectively), whereas most other elongator yeast tRNAs bear either m5C48 or m5C49, but never both in the same tRNA molecule | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + 5-methylcytosine40 in tRNA | - |
? | |
| S-adenosyl-L-methionine + cytosine40 in tRNA precursor | mini-tRNAPhe (composed of the anticodon stem-loop extended by the intron) is used to test the formation of m5C40. The formation of m5C40 is strictly intron dependent | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + 5-methylcytosine40 in tRNA precursor | - |
? | |
| S-adenosyl-L-methionine + cytosine48 in tRNA | the enzyme is responsible for complete m5C methylation of yeast tRNA. The frequency of modification depends on the cytosine position in tRNA. At positions 34 and 40, m5C is found only in two yeast tRNAs (tRNALeu (CUA) and tRNAPhe (GAA), respectively), whereas most other elongator yeast tRNAs bear either m5C48 or m5C49, but never both in the same tRNA molecule | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + 5-methylcytosine48 in tRNA | - |
? | |
| S-adenosyl-L-methionine + cytosine48 in tRNA | tRNATyr(GUA), tRNASer(AGA), and tRNAIle(UAU) are used to test the formation of m5C48 | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + 5-methylcytosine48 in tRNA | - |
? | |
| S-adenosyl-L-methionine + cytosine49 in tRNA | the enzyme is responsible for complete m5C methylation of yeast tRNA. The frequency of modification depends on the cytosine position in tRNA. At positions 34 and 40, m5C is found only in two yeast tRNAs (tRNALeu (CUA) and tRNAPhe (GAA), respectively), whereas most other elongator yeast tRNAs bear either m5C48 or m5C49, but never both in the same tRNA molecule | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + 5-methylcytosine49 in tRNA | - |
? | |
| S-adenosyl-L-methionine + cytosine49 in tRNA | tRNAPhe(GAA) and tRNAAsp(GUC) are used to test the formation of m5C49 | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + 5-methylcytosine49 in tRNA | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 77879, calculated from sequence | Saccharomyces cerevisiae |
| ? | x * 80200, calculated from sequence, including Hi26- and thrombin cleavage sequences | Saccharomyces cerevisiae |
| ? | x * 82200, SDS-PAGE, recombinant enzyme including Hi26- and thrombin cleavage sequences | Saccharomyces cerevisiae |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| multisite-specific tRNA:m5C-methyltransferase | - |
Saccharomyces cerevisiae |
| TRM4 | gene name, gene corresponding to ORF YBL024w | Saccharomyces cerevisiae |
| tRNA:m5C-methyltransferase | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Saccharomyces cerevisiae |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
assay at | Saccharomyces cerevisiae |
General Information
| General Information | Comment | Organism |
|---|---|---|
| malfunction | disruption of the ORF YBL024w leads to the complete absence of m5C in total yeast tRNA. No tRNA:m5C-methyltransferase activity towards all potential m5C methylation sites is detected in the extract of the disrupted yeast strain. The protein product of a single gene is responsible for complete m5C methylation of yeast tRNA | Saccharomyces cerevisiae |
| physiological function | the enzyme is responsible for complete m5C methylation of yeast tRNA | Saccharomyces cerevisiae |
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Release 2023.1 (January 2023)
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