BRENDA - Enzyme Database

Multisite-specific tRNA:m5C-methyltransferase (Trm4) in yeast Saccharomyces cerevisiae: identification of the gene and substrate specificity of the enzyme

Motorin, Y.; Grosjean, H.; RNA 5, 1105-1118 (1999)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.1.1.202
inserted in the expression vector pET28b, and the corresponding protein is hyperexpressed in Escherichia coli BL21(DE3)
Saccharomyces cerevisiae
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.1.1.202
77879
-
x * 77879, calculated from sequence
Saccharomyces cerevisiae
2.1.1.202
80200
-
x * 80200, calculated from sequence, including Hi26- and thrombin cleavage sequences
Saccharomyces cerevisiae
2.1.1.202
82200
-
x * 82200, SDS-PAGE, recombinant enzyme including Hi26- and thrombin cleavage sequences
Saccharomyces cerevisiae
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.1.1.202
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
-
-
?
2.1.1.202
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
-
-
?
2.1.1.202
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
-
-
?
2.1.1.202
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
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.1.1.202
Saccharomyces cerevisiae
P38205
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.1.1.202
recombinant Ybl024p is purified to apparent homogeneity by one-step affinity chromatography on Ni21-NTA-agarose column
Saccharomyces cerevisiae
Reaction
EC Number
Reaction
Commentary
Organism
2.1.1.202
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
2.1.1.202
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
2.1.1.202
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
2.1.1.202
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)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.202
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
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine34 in tRNA
-
-
-
?
2.1.1.202
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
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine34 in tRNA precursor
-
-
-
?
2.1.1.202
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
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine40 in tRNA
-
-
-
?
2.1.1.202
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
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine40 in tRNA precursor
-
-
-
?
2.1.1.202
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
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine48 in tRNA
-
-
-
?
2.1.1.202
S-adenosyl-L-methionine + cytosine48 in tRNA
tRNATyr(GUA), tRNASer(AGA), and tRNAIle(UAU) are used to test the formation of m5C48
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine48 in tRNA
-
-
-
?
2.1.1.202
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
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine49 in tRNA
-
-
-
?
2.1.1.202
S-adenosyl-L-methionine + cytosine49 in tRNA
tRNAPhe(GAA) and tRNAAsp(GUC) are used to test the formation of m5C49
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine49 in tRNA
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
2.1.1.202
?
x * 77879, calculated from sequence; x * 80200, calculated from sequence, including Hi26- and thrombin cleavage sequences; x * 82200, SDS-PAGE, recombinant enzyme including Hi26- and thrombin cleavage sequences
Saccharomyces cerevisiae
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.1.1.202
30
-
assay at
Saccharomyces cerevisiae
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.1.1.202
8
-
assay at
Saccharomyces cerevisiae
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.1.202
inserted in the expression vector pET28b, and the corresponding protein is hyperexpressed in Escherichia coli BL21(DE3)
Saccharomyces cerevisiae
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.1.1.202
77879
-
x * 77879, calculated from sequence
Saccharomyces cerevisiae
2.1.1.202
80200
-
x * 80200, calculated from sequence, including Hi26- and thrombin cleavage sequences
Saccharomyces cerevisiae
2.1.1.202
82200
-
x * 82200, SDS-PAGE, recombinant enzyme including Hi26- and thrombin cleavage sequences
Saccharomyces cerevisiae
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.1.1.202
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
-
-
?
2.1.1.202
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
-
-
?
2.1.1.202
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
-
-
?
2.1.1.202
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
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.1.202
recombinant Ybl024p is purified to apparent homogeneity by one-step affinity chromatography on Ni21-NTA-agarose column
Saccharomyces cerevisiae
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.202
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
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine34 in tRNA
-
-
-
?
2.1.1.202
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
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine34 in tRNA precursor
-
-
-
?
2.1.1.202
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
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine40 in tRNA
-
-
-
?
2.1.1.202
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
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine40 in tRNA precursor
-
-
-
?
2.1.1.202
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
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine48 in tRNA
-
-
-
?
2.1.1.202
S-adenosyl-L-methionine + cytosine48 in tRNA
tRNATyr(GUA), tRNASer(AGA), and tRNAIle(UAU) are used to test the formation of m5C48
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine48 in tRNA
-
-
-
?
2.1.1.202
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
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine49 in tRNA
-
-
-
?
2.1.1.202
S-adenosyl-L-methionine + cytosine49 in tRNA
tRNAPhe(GAA) and tRNAAsp(GUC) are used to test the formation of m5C49
639471
Saccharomyces cerevisiae
S-adenosyl-L-homocysteine + 5-methylcytosine49 in tRNA
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.1.1.202
?
x * 77879, calculated from sequence; x * 80200, calculated from sequence, including Hi26- and thrombin cleavage sequences; x * 82200, SDS-PAGE, recombinant enzyme including Hi26- and thrombin cleavage sequences
Saccharomyces cerevisiae
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.1.1.202
30
-
assay at
Saccharomyces cerevisiae
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.1.1.202
8
-
assay at
Saccharomyces cerevisiae
General Information
EC Number
General Information
Commentary
Organism
2.1.1.202
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
2.1.1.202
physiological function
the enzyme is responsible for complete m5C methylation of yeast tRNA
Saccharomyces cerevisiae
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.1.1.202
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
2.1.1.202
physiological function
the enzyme is responsible for complete m5C methylation of yeast tRNA
Saccharomyces cerevisiae