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Literature summary extracted from

  • Thomas, S.R.; Keller, C.A.; Szyk, A.; Cannon, J.R.; Laronde-Leblanc, N.A.
    Structural insight into the functional mechanism of Nep1/Emg1 N1-specific pseudouridine methyltransferase in ribosome biogenesis (2011), Nucleic Acids Res., 39, 2445-2457.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.260 expression of His6-tagged Nep1 in Escherichia coli strain RosettaTM-DE3-pLysS Saccharomyces cerevisiae
2.1.1.260 expression of wild-type and selenomethionine-labeled His6-tagged Nep1 in Escherichia coli strain RosettaTM-DE3-pLysS Archaeoglobus fulgidus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.1.260 recombinant detagged scNep1 dimer bound to S-adenosyl homocysteine and in complexes with RNA, i.e. one molecule and two molecules of cognate RNA, X-ray diffraction structure determination and analysis at 1.80-1.90 A and at 3.00 A resolution, respectively Saccharomyces cerevisiae
2.1.1.260 recombinant detagged wild-type and selenomethionine-labeled afNep1 dimer bound to S-adenosyl homocysteine, X-ray diffraction structure determination and analysis at 1.45-2.0 A resolution Archaeoglobus fulgidus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.1.1.260 nucleolus
-
 Saccharomyces cerevisiae 5730
-

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.260 S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA Saccharomyces cerevisiae
-
 S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?
2.1.1.260 S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA Archaeoglobus fulgidus
-
 S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.260 Archaeoglobus fulgidus O29524
-
-
2.1.1.260 Saccharomyces cerevisiae
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.260 recombinant His6-tagged Nep1 from Escherichia coli strain RosettaTM-DE3-pLysS by nickel affinity chromatography, cleavage of the tag by TEV protease, and two steps of gel filtration to over 99% purity Saccharomyces cerevisiae
2.1.1.260 recombinant wild-type and selenomethionine-labeled His6-tagged Nep1 from Escherichia coli strain RosettaTM-DE3-pLysS by nickel affinity chromatography, cleavage of the tag by TEV protease, and two steps of gel filtration to over 99% purity Archaeoglobus fulgidus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.260 S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA
-
 Saccharomyces cerevisiae S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?
2.1.1.260 S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA
-
 Archaeoglobus fulgidus S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?
2.1.1.260 S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA S-adenosyl-L-methionine binding structure, overview Saccharomyces cerevisiae S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA S-adenosyl-L-homocysteine binding structure, overview ?
2.1.1.260 S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA S-adenosyl-L-methionine binding structure, overview Archaeoglobus fulgidus S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA S-adenosyl-L-homocysteine binding structure, overview ?

Subunits

EC Number Subunits Comment Organism
2.1.1.260 dimer
-
 Archaeoglobus fulgidus
2.1.1.260 dimer structure of the dimerization interface of scNep1, overview Saccharomyces cerevisiae

Synonyms

EC Number Synonyms Comment Organism
2.1.1.260 Emg1
-
 Saccharomyces cerevisiae
2.1.1.260 Emg1
-
 Archaeoglobus fulgidus
2.1.1.260 N1-specific pseudouridine methyltransferase
-
 Saccharomyces cerevisiae
2.1.1.260 N1-specific pseudouridine methyltransferase
-
 Archaeoglobus fulgidus
2.1.1.260 Nep1
-
 Saccharomyces cerevisiae
2.1.1.260 Nep1
-
 Archaeoglobus fulgidus
2.1.1.260 pseudouridine N1-methyltransferase
-
 Saccharomyces cerevisiae
2.1.1.260 pseudouridine N1-methyltransferase
-
 Archaeoglobus fulgidus

General Information

EC Number General Information Comment Organism
2.1.1.260 malfunction mutations in Nep1 result in decreased methyl donor binding, but do not result in lethality Saccharomyces cerevisiae
2.1.1.260 additional information active site structure of scNep1, overview Saccharomyces cerevisiae
2.1.1.260 physiological function Nep1 is a SPOUT RNA methyltransferase, and can catalyze methylation at the N1 of pseudouridine, it is required for 18S rRNA maturation. Nep1 is also involved in assembly of Rps19, an SSU ribosomal protein. Functional mechanism of Nep1/Emg1 N1-specific pseudouridine methyltransferase in ribosome biogenesis, structure-function relationship, overview Archaeoglobus fulgidus
2.1.1.260 physiological function Nep1 is a SPOUT RNA methyltransferase, and can catalyze methylation at the N1 of pseudouridine, it is required for 18S rRNA maturation. Nep1 is also involved in assembly of Rps19, an SSU ribosomal protein. Functional mechanism of Nep1/Emg1 N1-specific pseudouridine methyltransferase in ribosome biogenesis, structure-function relationship, overview. Nep1 recognizes its RNA site via base-specific interactions and stabilizes a stem-loop in the bound RNA. Nep1 changes rRNA structure upon binding, a uridine base is bound in the active site of Nep1, positioned for a methyltransfer at the C5 position Saccharomyces cerevisiae