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Literature summary for 3.1.26.3 extracted from

  • Nagel, R.; Ares, M.Jr.
    Substrate recognition by a eukaryotic RNase III: the double-stranded RNA-binding domain of Rnt1p selectively binds RNA containing a 5'-AGNN-3' tetraloop (2000), RNA, 6, 1142-1156.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli BL21(DE3) as C-terminally His-tagged proteins Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
additional information construction of deletion mutants RNT1DELTA2-329 and RNT1DELTA2-198, which are both catalytically active in vitro but do not rescue a growth defective mutant and are not able to retain activity and viability in vivo, construction of a AGNN-loop exchange mutant GNRA-loop shows reduced activity and substrate selectivity Saccharomyces cerevisiae

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Saccharomyces cerevisiae

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
55000
-
2 * 55000, amino acid sequence calculation Saccharomyces cerevisiae
110000
-
recombinant wild-type enzyme, gel filtration Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
pre-rRNA + H2O Saccharomyces cerevisiae enzyme is required for processing mature rRNA
-
?
pre-snoRNA + H2O Saccharomyces cerevisiae enzyme is required for processing mature snoRNA
-
?
pre-snRNA + H2O Saccharomyces cerevisiae enzyme is required for processing mature snRNA
-
?

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli, expression of the mutant enzymes in a two-hybrid system in Saccharomyces strains Saccharomyces cerevisiae

Reaction

Reaction Comment Organism Reaction ID
Endonucleolytic cleavage to a 5'-phosphomonoester RNA substrate forms a duplex structure with a terminal four-base loop with the sequence AGNN, being an important determinant of the substrate selectivity in the dsRNA binding domain dsRBD, the N-terminal region is not required for selective cleavage in vitro but required for full function and viability of yeast cells in vivo, substrate binding modeling Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
pre-rRNA + H2O
-
Saccharomyces cerevisiae mature rRNA
-
?
pre-rRNA + H2O enzyme is required for processing Saccharomyces cerevisiae mature rRNA
-
?
pre-snoRNA + H2O
-
Saccharomyces cerevisiae mature snoRNA
-
?
pre-snoRNA + H2O enzyme is required for processing Saccharomyces cerevisiae mature snoRNA
-
?
pre-snRNA + H2O
-
Saccharomyces cerevisiae mature snRNA
-
?
pre-snRNA + H2O enzyme is required for processing Saccharomyces cerevisiae mature snRNA
-
?
synthetic 25S rRNA 3' ETS cleavage site containing RNA + H2O
-
Saccharomyces cerevisiae ?
-
?

Subunits

Subunits Comment Organism
dimer 2 * 55000, amino acid sequence calculation Saccharomyces cerevisiae
More deletion mutant RNT1DELTA2-198 also behaves as a dimer Saccharomyces cerevisiae

Synonyms

Synonyms Comment Organism
RNase III
-
Saccharomyces cerevisiae
RNT1
-
Saccharomyces cerevisiae
Rnt1p
-
Saccharomyces cerevisiae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Saccharomyces cerevisiae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Saccharomyces cerevisiae