Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant enzymes | Saccharomyces cerevisiae |
Crystallization (Comment) | Organism |
---|---|
analysis of crystal structures of Bud23-Trm112 MTase complexes, overview | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
D112A | site-directed mutagenesis | Saccharomyces cerevisiae |
D112R | site-directed mutagenesis, methylation inactive mutant | Saccharomyces cerevisiae |
D77A | site-directed mutagenesis, methylation inactive mutant, 25% remaining S-adenosyl-L-methionine binding activity | Saccharomyces cerevisiae |
D94A | site-directed mutagenesis | Saccharomyces cerevisiae |
D94R | site-directed mutagenesis | Saccharomyces cerevisiae |
E18A | site-directed mutagenesis | Saccharomyces cerevisiae |
I31W | site-directed mutagenesis, methylation inactive mutant, 25% remaining S-adenosyl-L-methionine binding activity | Saccharomyces cerevisiae |
K21E/R27E | site-directed mutagenesis, methylation inactive mutant | Saccharomyces cerevisiae |
M96A | site-directed mutagenesis | Saccharomyces cerevisiae |
additional information | protein levels and cell growth of mutants compared to wild-type, overview. Reconstitution of Dhr1-Bud23-Trm112 ternary complex by mixing Bud23-Trm112 with a 1.5 M excess of Dhr1[58-270] in 20 mM Tris·HCl, pH 7.5, 50 mM NaCl, 0.010 mM ZnCl2, 5 mM 2-mercaptoethanol | Saccharomyces cerevisiae |
S118E | site-directed mutagenesis, methylation and S-adenosyl-L-methionine binding inactive mutant | Saccharomyces cerevisiae |
S118R | site-directed mutagenesis, methylation and S-adenosyl-L-methionine binding inactive mutant | Saccharomyces cerevisiae |
W122A | site-directed mutagenesis, methylation inactive mutant | Saccharomyces cerevisiae |
Y159A | site-directed mutagenesis, methylation inactive mutant, 50% remaining S-adenosyl-L-methionine binding activity | Saccharomyces cerevisiae |
Y22A | site-directed mutagenesis, methylation inactive mutant | Saccharomyces cerevisiae |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | required | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Saccharomyces cerevisiae | Bud23-Trm112 interacts directly with the box C/D snoRNA U3-associated DEAH RNA helicase Dhr1 supposedly involved in central pseudoknot formation. Bud23Trm112 might also contribute to controlling formation of this irreversible and dramatic structural reorganization essential to overall folding of small subunit rRNA | ? | - |
? | |
S-adenosyl-L-methionine + guanine1575 in 18S rRNA | Saccharomyces cerevisiae | Bud23 is active in 18S rRNA N7G methylation as a heterodimer in a complex with Trm112. Trm112 is a small zinc finger protein that interacts with and activates three class I MTases in addition to Bud23. N7-methylguanosine introduced at position 1575 on 18S rRNA by Bud23-Trm112 is at a ridge forming a steric block between P- and E-site tRNAs. G1575 is coordinated by a network of conserved Bud23 residues, overview | S-adenosyl-L-homocysteine + N7-methylguanine1575 in 18S rRNA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | P25627 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Bud23-Trm112 interacts directly with the box C/D snoRNA U3-associated DEAH RNA helicase Dhr1 supposedly involved in central pseudoknot formation. Bud23Trm112 might also contribute to controlling formation of this irreversible and dramatic structural reorganization essential to overall folding of small subunit rRNA | Saccharomyces cerevisiae | ? | - |
? | |
S-adenosyl-L-methionine + guanine1575 in 18S rRNA | Bud23 is active in 18S rRNA N7G methylation as a heterodimer in a complex with Trm112. Trm112 is a small zinc finger protein that interacts with and activates three class I MTases in addition to Bud23. N7-methylguanosine introduced at position 1575 on 18S rRNA by Bud23-Trm112 is at a ridge forming a steric block between P- and E-site tRNAs. G1575 is coordinated by a network of conserved Bud23 residues, overview | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + N7-methylguanine1575 in 18S rRNA | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BUD23 | - |
Saccharomyces cerevisiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Saccharomyces cerevisiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Saccharomyces cerevisiae |
General Information | Comment | Organism |
---|---|---|
additional information | Bud23 and Trm112 interact through formation of a beta-zipper involving main-chain atoms, burying an important hydrophobic surface and stabilizing the complex. The structures reveal that the coactivator Trm112 undergoes an induced fit to accommodate its methyltransferase (MTase) partner, identification of Bud23 residues important for Bud23-Trm112 complex formation and recruitment to pre-ribosomes | Saccharomyces cerevisiae |
physiological function | Bud23-Trm112 is required for efficient pre-rRNA processing steps leading to 18S rRNA synthesis and methylation of 18S rRNA at position G1575. Identification amd localization of Bud23Trm112 contacts with precursor ribosomes. The essential helicase Dhr1 interacts directly with Bud23-Trm112, proposing a concerted action of these proteins in ribosome assembly. The methyltransferase activity of Bud23-Trm112 is required for pre-rRNA processing are disconnected in time. Though Bud23-Trm112 binds precursor ribosomes at an early nucleolar stage, m7G methylation occurs at a late step of small subunit biogenesis, implying specifically delayed catalytic activation. Bud23-Trm112 interacts directly with the box C/D snoRNA U3-associated DEAH RNA helicase Dhr1 supposedly involved in central pseudoknot formation suggesting that Bud23-Trm112 might also contribute to controlling formation of this irreversible and dramatic structural reorganization essential to overall folding of small subunit rRNA. Formation of the Bud23-Trm112 complex is required for efficient 18S rRNA maturation | Saccharomyces cerevisiae |