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Literature summary for 2.1.1.260 extracted from
- The Bowen-Conradi syndrome protein Nep1 (Emg1) has a dual role in eukaryotic ribosome biogenesis, as an essential assembly factor and in the methylation of psi1191 in yeast 18S rRNA (2011), Nucleic Acids Res., 39, 1526-1537.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene NEP1, expression of N-terminally His6-tagged wild-type and D90G mutant Nep1 in Escherichia coli strain XL1-blue | Saccharomyces cerevisiae |
| gene NEP1, stable expression of GFP-tagged wild-type and D86G mutant Nep1 in 293T cells under control of the tetracycline-inducible promoter, expression of N-terminally His6-tagged wild-type and D90G mutant Nep1 in Escherichia coli strain XL1-blue | Homo sapiens |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D90G | a mutation in yeast Nep1 equivalent to the Bowen-Conradi syndrome, BCS, mutation in the human Nep1. Nep1 methyltransferase activity is not affected upon introduction of the BCS mutation, the mutated protein shows enhanced dimerization propensity and increased affinity for its RNA-target in vitro | Saccharomyces cerevisiae |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| nucleolus | - |
Homo sapiens | 5730 | - |
| nucleolus | - |
Saccharomyces cerevisiae | 5730 | - |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 32000 | - |
x * 32000, His6-tagged Nep1, SD-PAGE | Saccharomyces cerevisiae |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Saccharomyces cerevisiae | Nep1 target site identification by mass spectrometry, isotope-labeling, and fluorescence anisotropy measurements | ? | - |
? |  |
| S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | Homo sapiens | - |
S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | - |
? | |
| S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | Saccharomyces cerevisiae | Nep1 catalyzes the psi1191 methylation in vivo. The in vivo target site for Nep1-catalyzed methylation is located within loop 35 of the 18S rRNA that contains the unique hypermodification of U1191 to 1-methyl-3-(3-amino-3-carboxypropyl)-pseudouridine | S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Homo sapiens | - |
gene NEP1 | - |
| Saccharomyces cerevisiae | - |
gene NEP1 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant N-terminally His6-tagged wild-type and D90G mutant Nep1 from Escherichia coli strain XL1-blue by nickel affinity chromatography and gel filtration | Homo sapiens |
| recombinant N-terminally His6-tagged wild-type and D90G mutant Nep1 from Escherichia coli strain XL1-blue by nickel affinity chromatography and gel filtration | Saccharomyces cerevisiae |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Nep1 target site identification by mass spectrometry, isotope-labeling, and fluorescence anisotropy measurements | Saccharomyces cerevisiae | ? | - |
? | |
| S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | - |
Homo sapiens | S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | - |
? | |
| S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | - |
Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | - |
? | |
| S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA | Nep1 catalyzes the psi1191 methylation in vivo. The in vivo target site for Nep1-catalyzed methylation is located within loop 35 of the 18S rRNA that contains the unique hypermodification of U1191 to 1-methyl-3-(3-amino-3-carboxypropyl)-pseudouridine | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 32000, His6-tagged Nep1, SD-PAGE | Saccharomyces cerevisiae |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Bowen-Conradi syndrome protein | - |
Homo sapiens |
| Bowen-Conradi syndrome protein | - |
Saccharomyces cerevisiae |
| Emg1 | - |
Homo sapiens |
| Emg1 | - |
Saccharomyces cerevisiae |
| Nep1 | - |
Homo sapiens |
| Nep1 | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Homo sapiens |
| 37 | - |
assay at | Saccharomyces cerevisiae |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
assay at | Homo sapiens |
| 7.5 | - |
assay at | Saccharomyces cerevisiae |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| S-adenosyl-L-methionine | - |
Homo sapiens | |
| S-adenosyl-L-methionine | - |
Saccharomyces cerevisiae | |
Expression
| Organism | Comment | Expression |
|---|---|---|
| Saccharomyces cerevisiae | S-adenosylmethionine suppression of Nep1, suppression of mutant ScDELTAsnr35 nep1-1ts is independent of psi1191 methylation | down |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | Nep1 is a member of the SPOUT-family of methyltransferases | Homo sapiens |
| evolution | Nep1 is a member of the SPOUT-family of methyltransferases | Saccharomyces cerevisiae |
| malfunction | Nep1 mutation D86G causes the Bowen-Conradi syndrome, BCS, that results in severe pre and postnatal growth and psychomotor retardation, microcephaly, micrognathia, rocker bottom feet and early childhood death, overview. Human HsNep1D86G protein shows a strongly increased interaction of the monomers | Homo sapiens |
| malfunction | restored growth of a nep1-1ts mutant upon addition of S-adenosylmethionine also after preventing U1191 methylation in a DElTAsnr35 mutant. Nep1 methyltransferase activity is not affected upon introduction of the Bowen-Conradi syndrome, BCS, D86G mutation. Instead, the mutated protein shows enhanced dimerization propensity and increased affinity for its RNA-target in vitro | Saccharomyces cerevisiae |
| metabolism | replacement of U1191 by any other base caused significant growth deficiencies | Saccharomyces cerevisiae |
| physiological function | Nep1 is essential for psi119 pseudouridine methylation, but is not required for acp-modification. Nep1 has a dual function, as psi1191-methyltransferase and ribosome assembly factor | Saccharomyces cerevisiae |
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