Cloned (Comment) | Organism |
---|---|
gene trm7, recombinant expression of trm7 in Escherichia coli strain BL21(DE3) Rosetta 2, coexpression of trm732 and trm734, construction of Trm7-Trm734 and Trm7-Trm732 co-expression vectors | Saccharomyces cerevisiae |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme complexes Trm7-Trm734 and Trm7-Trm734-SAM, hanging drop vapor diffusion technique, mixing of 3 mg/ml protein solution with reservoir solution containing 0.1 M HEPES-NaOH, pH 7.5, 2 M (NH4)2SO4, 2% PEG 400, 2 mM phenol, and 3% ethylene-glycol, and equilibration against 0.2m ml reservoir solution, soaking of the crystals in in the mother liquor containing 1 mM SAM for complex 2, 20°C, 5 days, X-ray diffraction structure determination and analysis at 2.32-3.20 A resolution | Saccharomyces cerevisiae |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytoplasm | Trm7, Trm732, and Trm734 are localized to cytoplasm | Saccharomyces cerevisiae | 5737 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + cytidine32/guanosine34 in tRNALeu | Saccharomyces cerevisiae | enzyme complex Trm7-Trm734 | S-adenosyl-L-homocysteine + 2'-O-methylcytidine32/2'-O methylguanosine34 in tRNALeu | - |
? | |
S-adenosyl-L-methionine + cytidine32/guanosine34 in tRNALeu | Saccharomyces cerevisiae ATCC 204508 | enzyme complex Trm7-Trm734 | S-adenosyl-L-homocysteine + 2'-O-methylcytidine32/2'-O methylguanosine34 in tRNALeu | - |
? | |
S-adenosyl-L-methionine + cytidine32/guanosine34 in tRNAPhe | Saccharomyces cerevisiae | enzyme complex Trm7-Trm734 | S-adenosyl-L-homocysteine + 2'-O-methylcytidine32/2'-O methylguanosine34 in tRNAPhe | - |
? | |
S-adenosyl-L-methionine + cytidine32/guanosine34 in tRNAPhe | Saccharomyces cerevisiae ATCC 204508 | enzyme complex Trm7-Trm734 | S-adenosyl-L-homocysteine + 2'-O-methylcytidine32/2'-O methylguanosine34 in tRNAPhe | - |
? | |
S-adenosyl-L-methionine + cytidine32/guanosine34 in tRNATrp | Saccharomyces cerevisiae | enzyme complex Trm7-Trm734 | S-adenosyl-L-homocysteine + 2'-O-methylcytidine32/2'-O methylguanosine34 in tRNATrp | - |
? | |
S-adenosyl-L-methionine + cytidine32/guanosine34 in tRNATrp | Saccharomyces cerevisiae ATCC 204508 | enzyme complex Trm7-Trm734 | S-adenosyl-L-homocysteine + 2'-O-methylcytidine32/2'-O methylguanosine34 in tRNATrp | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | P38238 | - |
- |
Saccharomyces cerevisiae ATCC 204508 | P38238 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzymatic activity of Trm7-Trm734 is measured by incorporation of 14C-methyl groups from [methyl-14C]-SAM into tRNA transcripts. Although these mutant tRNAPhe transcripts are methylated by Trm7-Trm734, the substitution of G34 by C or U (G34C or G34U) strongly enhances the methyl-group acceptance activity. Trm7-Trm734 preferentially methylates pyrimidine nucleoside at position 34 (pyrimidine34) rather than purine nucleoside. Of the 43 tRNA molecular species in Saccharomyces cerevisiae, only tRNAPhe, tRNATrp and tRNALeu possess the combination of Cm32, m1G37 and/or pyrimidine34 in the normal anticodon-loop, which is required for the effective methylation by Trm7-Trm734 | Saccharomyces cerevisiae | ? | - |
- |
|
additional information | the enzymatic activity of Trm7-Trm734 is measured by incorporation of 14C-methyl groups from [methyl-14C]-SAM into tRNA transcripts. Although these mutant tRNAPhe transcripts are methylated by Trm7-Trm734, the substitution of G34 by C or U (G34C or G34U) strongly enhances the methyl-group acceptance activity. Trm7-Trm734 preferentially methylates pyrimidine nucleoside at position 34 (pyrimidine34) rather than purine nucleoside. Of the 43 tRNA molecular species in Saccharomyces cerevisiae, only tRNAPhe, tRNATrp and tRNALeu possess the combination of Cm32, m1G37 and/or pyrimidine34 in the normal anticodon-loop, which is required for the effective methylation by Trm7-Trm734 | Saccharomyces cerevisiae ATCC 204508 | ? | - |
- |
|
S-adenosyl-L-methionine + cytidine32/guanosine34 in tRNALeu | enzyme complex Trm7-Trm734 | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + 2'-O-methylcytidine32/2'-O methylguanosine34 in tRNALeu | - |
? | |
S-adenosyl-L-methionine + cytidine32/guanosine34 in tRNALeu | enzyme complex Trm7-Trm734 | Saccharomyces cerevisiae ATCC 204508 | S-adenosyl-L-homocysteine + 2'-O-methylcytidine32/2'-O methylguanosine34 in tRNALeu | - |
? | |
S-adenosyl-L-methionine + cytidine32/guanosine34 in tRNAPhe | enzyme complex Trm7-Trm734 | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + 2'-O-methylcytidine32/2'-O methylguanosine34 in tRNAPhe | - |
? | |
S-adenosyl-L-methionine + cytidine32/guanosine34 in tRNAPhe | enzyme complex Trm7-Trm734 | Saccharomyces cerevisiae ATCC 204508 | S-adenosyl-L-homocysteine + 2'-O-methylcytidine32/2'-O methylguanosine34 in tRNAPhe | - |
? | |
S-adenosyl-L-methionine + cytidine32/guanosine34 in tRNATrp | enzyme complex Trm7-Trm734 | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + 2'-O-methylcytidine32/2'-O methylguanosine34 in tRNATrp | - |
? | |
S-adenosyl-L-methionine + cytidine32/guanosine34 in tRNATrp | enzyme complex Trm7-Trm734 | Saccharomyces cerevisiae ATCC 204508 | S-adenosyl-L-homocysteine + 2'-O-methylcytidine32/2'-O methylguanosine34 in tRNATrp | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterodimer | Trm7 forms a heterodimer with Trm734 | Saccharomyces cerevisiae |
Synonyms | Comment | Organism |
---|---|---|
trm7 | - |
Saccharomyces cerevisiae |
Trm7p | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Saccharomyces cerevisiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Saccharomyces cerevisiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | binding structure with Trm7-Trm734, overview | Saccharomyces cerevisiae |
General Information | Comment | Organism |
---|---|---|
evolution | sequence and structure-function comaprisons of yeast Trm7 and Trm7 enzymes from other species. Trm7 has the class I-type Rossmann fold, frequently found in SAM-dependent RNA methyltransferases, and probably employs a 2'-O-methylation mechanism similar to that of Escherichia coli RlmE, given that their overall structures are similar and that catalytic residues and SAM-binding mode are highly conserved | Saccharomyces cerevisiae |
metabolism | the degree of some of the modifications in tRNAs from Saccharomyces cerevisiae may be regulated by the balance between substrate tRNAs and modification enzymes. Because one of key modifications, Cm32, is conferred by Trm7-Trm732, it is clear that the activity of Trm7-Trm734 is regulated by the quantitative balance between Trm732 and Trm734 | Saccharomyces cerevisiae |
additional information | the structure of tRNA methyltransferase complex of Trm7 and Trm734 (regulator of Ty1 transposition protein 10, RTT10, UniProt Q08924) reveals another binding interface for tRNA recognition, crystal structure analysis and substrate tRNA recognition by Trm7-Trm734, and active site structure, overview. Hydrogen bonds and salt bridges between Trm7 and Trm734 are determined | Saccharomyces cerevisiae |