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Literature summary for 2.1.1.260 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
Search for more literature for 2.1.1.260

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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
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

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

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 Saccharomyces cerevisiae
-
 S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?
S-adenosyl-L-methionine + pseudouridine1191 in yeast 18S rRNA Archaeoglobus fulgidus
-
 S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?

Organism

Organism UniProt Comment Textmining
Archaeoglobus fulgidus O29524
-
-
Saccharomyces cerevisiae
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
 Archaeoglobus fulgidus S-adenosyl-L-homocysteine + N1-methylpseudouridine1191 in yeast 18S rRNA
-
 ?
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 ?
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

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

Synonyms

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

General Information

General Information Comment Organism
malfunction mutations in Nep1 result in decreased methyl donor binding, but do not result in lethality Saccharomyces cerevisiae
additional information active site structure of scNep1, overview Saccharomyces cerevisiae
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
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