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Literature summary for 3.1.26.3 extracted from

  • Cenik, E.S.; Fukunaga, R.; Lu, G.; Dutcher, R.; Wang, Y.; Tanaka Hall, T.M.; Zamore, P.D.
    Phosphate and R2D2 restrict the substrate specificity of Dicer-2, an ATP-driven ribonuclease (2011), Mol. Cell, 42, 172-184.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.1.26.3

Activating Compound

Activating Compound Comment Organism Structure
ATP ATP hydrolysis is required for Dicer-2 to process long dsRNA, but not pre-miRNA Drosophila melanogaster
dsRNA activates Dicer-2 Drosophila melanogaster
Loquacious-PB activates dsRNA cleavage of Dicer-2 Drosophila melanogaster
protein R2D2 Dicer-2 partner protein R2D2 inhibit pre-miRNA cleavage in vivo by Dicer-2, but activates dsRNA cleavage of Dicer-2 Drosophila melanogaster

Cloned(Commentary)

Cloned (Comment) Organism
expression of His6-tagged wild-type and mutant Dicer-2 in Spodoptera frugiperda Sf21 cells Drosophila melanogaster

Protein Variants

Protein Variants Comment Organism
D1217N/D1614N site-directed mutagenesis of Dicer-2 Drosophila melanogaster
G31R site-directed mutagenesis of Dicer-2 Drosophila melanogaster

Inhibitors

Inhibitors Comment Organism Structure
phosphate inhibits Dicer-2 Drosophila melanogaster
protein R2D2 Dicer-2 partner protein R2D2 inhibit pre-miRNA cleavage in vivo by Dicer-2, but activates dsRNA cleavage of Dicer-2 Drosophila melanogaster

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten Analysis of Dicer-2 Incubated with a 515 bp dsRNA and ATP, overview Drosophila melanogaster
0.0000004
-
 515 bp dsRNA wild-type Dicer-2 in presence of Loquacious-PB at equimolar level, pH 7.4, 25°C Drosophila melanogaster
0.000002
-
 515 bp dsRNA wild-type Dicer-2 in presence of protein R2D2 at equimolar level, pH 7.4, 25°C Drosophila melanogaster
0.000006
-
 515 bp dsRNA wild-type Dicer-2, pH 7.4, 25°C Drosophila melanogaster

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Drosophila melanogaster Dicer-1 and Dicer-2 show different substrate specificity in vivo: Dicer-2 generates small interfering RNAs, siRNAs, from long double-stranded RNA, dsRNA, whereas Dicer-1 produces microRNAs, miRNAs, from pre-miRNA. Dicer-2 can efficiently cleave pre-miRNA in vitro, but phosphate and the Dicer-2 partner protein R2D2 inhibit pre-miRNA cleavage in vivo. Wild-type Dicer-2, but not a mutant defective in ATP hydrolysis, can generate siRNAs faster than it can dissociate from a long dsRNA substrate. Dicer-1 does not efficiently process long dsRNA ?
-
 ?

Organism

Organism UniProt Comment Textmining
Drosophila melanogaster
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged wild-type and mutant Dicer-2 from Sf21 insect cells by nickel affinity and heparin affinity chromatography, followed by gel filtration Drosophila melanogaster

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
515 bp dsRNA + H2O Dicer-2 substrate is synthetic 515 bp dsRNA Drosophila melanogaster ?
-
 ?
additional information Dicer-1 and Dicer-2 show different substrate specificity in vivo: Dicer-2 generates small interfering RNAs, siRNAs, from long double-stranded RNA, dsRNA, whereas Dicer-1 produces microRNAs, miRNAs, from pre-miRNA. Dicer-2 can efficiently cleave pre-miRNA in vitro, but phosphate and the Dicer-2 partner protein R2D2 inhibit pre-miRNA cleavage in vivo. Wild-type Dicer-2, but not a mutant defective in ATP hydrolysis, can generate siRNAs faster than it can dissociate from a long dsRNA substrate. Dicer-1 does not efficiently process long dsRNA Drosophila melanogaster ?
-
 ?

Subunits

Subunits Comment Organism
More Dicer-2 contains C-terminal RNase III domains, that mediate RNA cleavage, and an N-terminal helicase motif Drosophila melanogaster

Synonyms

Synonyms Comment Organism
Dicer-2
-
 Drosophila melanogaster

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Drosophila melanogaster

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0005
-
 515 bp dsRNA wild-type Dicer-2 in presence of Loquacious-PB at equimolar level, pH 7.4, 25°C Drosophila melanogaster
0.0005
-
 515 bp dsRNA wild-type Dicer-2 in presence of protein R2D2 at equimolar level, pH 7.4, 25°C Drosophila melanogaster
0.0005
-
 515 bp dsRNA wild-type Dicer-2, pH 7.4, 25°C Drosophila melanogaster

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
 assay at Drosophila melanogaster

General Information

General Information Comment Organism
additional information Dicer-2 contains C-terminal RNase III domains that mediate RNA cleavage Drosophila melanogaster
physiological function Dicer-2 is a dsRNA-stimulated ATPase that hydrolyzes ATP to ADP. Dicer-2 generates small interfering RNAs, siRNAs, from long double-stranded RNA, dsRNA Drosophila melanogaster

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.083
-
 515 bp dsRNA wild-type Dicer-2, pH 7.4, 25°C Drosophila melanogaster
0.33
-
 515 bp dsRNA wild-type Dicer-2 in presence of protein R2D2 at equimolar level, pH 7.4, 25°C Drosophila melanogaster
1.33
-
 515 bp dsRNA wild-type Dicer-2 in presence of Loquacious-PB at equimolar level, pH 7.4, 25°C Drosophila melanogaster