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Literature summary for 2.1.1.260 extracted from

  • Wurm, J.; Meyer, B.; Bahr, U.; Held, M.; Frolow, O.; Koetter, P.; Engels, J.; Heckel, A.; Karas, M.; Entian, K.; Woehnert, J.
    The ribosome assembly factor Nep1 responsible for Bowen-Conradi syndrome is a pseudouridine-N1-specific methyltransferase (2010), Nucleic Acids Res., 38, 2387-2398.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.1.1.260

Cloned(Commentary)

Cloned (Comment) Organism
gene NEP1, recombinaant expression of Nep1 Methanocaldococcus jannaschii
gene NEP1, recombinant expression of His-tagged Nep1 in Escherichia coli strain M15(pREP4) Homo sapiens

Protein Variants

Protein Variants Comment Organism
D86G the mutation causes the Bowen-Conradi syndrome Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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 Methanocaldococcus jannaschii
-
 S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-
Methanocaldococcus jannaschii
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged Nep1 from Escherichia coli strain M15(pREP4) by nickel affinity chromatography Homo sapiens
recombinant Nep1 by cation exchange chromatography and gel filtration Methanocaldococcus jannaschii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information RNA-binding specificity by in vitro using fluorescence quenching assays and yeast three-hybrid screening. Identification of the binding site for methylation target RNAs by NMR-spectroscopy, overview Homo sapiens ?
-
 ?
additional information the Methanocaldococcus jannaschii Nep1 binds to the Saccharomyces cerevisiae Nep1 RNA consensus sequence 5'-UUCAAC-3' in the Methanocaldococcus jannaschii 16S rRNA. The region of Methanocaldococcus jannaschii 16S rRNA is equivalent to nt 1190-1195 in yeast 18S rRNA including the m1acp3-psi nucleotide at position 1191. And binding of MjNep1 to a 6mer RNA containing the consensus sequence 5'-UUCAAC-3'. Even a 5'-truncated RNA lacking the first uridine of the consensus sequence is bound with comparable affinity. RNA-binding specificity by in vitro using fluorescence quenching assays and yeast three-hybrid screening. Identification of the binding site for methylation target RNAs by NMR-spectroscopy, overview Methanocaldococcus jannaschii ?
-
 ?
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
-
 Methanocaldococcus jannaschii S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?
S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA
-
 Methanocaldococcus jannaschii S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA product identification by 1H-NMR-spectroscopy ?
S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA RNA-oligonucleotides corresponding to nucleotides 1245-1255 of human 18S rRNA and containing pseudouridine (5'-GACWCAACACG-3') at position 1248. This position corresponds to nucleotide 914 of Methanocaldococcus jannaschii 16S rRNA and to nt 1191 of yeast 18S rRNA Homo sapiens S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA product identification by MALDI-mass spectrometrical analysis ?

Subunits

Subunits Comment Organism
dimer three-dimensional structure of the Nep1 dimer, overview Homo sapiens
dimer three-dimensional structure of the Nep1 dimer, overview Methanocaldococcus jannaschii

Synonyms

Synonyms Comment Organism
Emg1
-
 Homo sapiens
Emg1
-
 Methanocaldococcus jannaschii
Nep1
-
 Homo sapiens
Nep1
-
 Methanocaldococcus jannaschii
Nep1 methyltransferase
-
 Homo sapiens
Nep1 methyltransferase
-
 Methanocaldococcus jannaschii
pseudouridine-N1-specific methyltransferase
-
 Homo sapiens
pseudouridine-N1-specific methyltransferase
-
 Methanocaldococcus jannaschii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Homo sapiens
65
-
 assay at Methanocaldococcus jannaschii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Homo sapiens
7.5
-
 assay at Methanocaldococcus jannaschii

Cofactor

Cofactor Comment Organism Structure
S-adenosyl-L-methionine
-
 Homo sapiens
S-adenosyl-L-methionine
-
 Methanocaldococcus jannaschii

General Information

General Information Comment Organism
evolution Nep1 belongs to the SPOUT-class RNA methyltransferases, Nep1 subfamily Homo sapiens
evolution Nep1 belongs to the SPOUT-class RNA methyltransferases, Nep1 subfamily Methanocaldococcus jannaschii
physiological function MjNep1 binds its RNA target along an extended basic surface cleft at the dimer interface that involves both insertions to the SPOUT-class core fold in an orientation consistent with the proposed methylation of nt 914 in Methanocaldococcus jannaschii 16S rRNA Methanocaldococcus jannaschii