Datasheet

生物学的記述

Specificity	Drosha Antibody (Rabbit mAb) [M15H18] detects endogenous levels of total Drosha proteins.
Background	Drosha is the catalytic ribonuclease III enzyme of the Microprocessor complex, functioning together with its obligate RNA-binding partner DGCR8 to catalyze the first, nuclear step of microRNA biogenesis: cleavage of the hairpin structures embedded within long primary microRNA transcripts (pri-miRNAs) to release the shorter precursor miRNA (pre-miRNA) that is subsequently exported to the cytoplasm for further processing by Dicer. Cryo-electron microscopy structures of the Drosha-DGCR8 complex, solved both with and without a bound pri-miRNA, show that a helix-hairpin element within an extended PAZ domain of Drosha, together with a mobile basic helix in Drosha's RNase IIIa domain, jointly recognize the single-stranded-to-double-stranded junction of the pri-miRNA hairpin, while Drosha's double-stranded RNA-binding domain makes extensive contacts along the RNA stem; the RNA-free structure additionally reveals that this same PAZ helix-hairpin element adopts an autoinhibitory conformation in the absence of substrate, indicating that pri-miRNA binding itself triggers a conformational change required to position Drosha's catalytic centers for correct, sequence-independent cleavage site selection. DGCR8 is required not only for RNA substrate recognition through its own double-stranded RNA-binding domains but also for stabilizing Drosha itself: DGCR8 positively regulates Drosha protein levels through a direct protein-protein interaction, while Drosha reciprocally regulates DGCR8 by cleaving conserved hairpin structures embedded within the DGCR8 mRNA itself, a Microprocessor-mediated cleavage event that destabilizes the DGCR8 transcript and reduces DGCR8 protein output. This reciprocal, cross-regulatory relationship, in which Drosha stabilizes its partner at the protein level while DGCR8 destabilizes Drosha's partner at the mRNA level, establishes a homeostatic feedback loop that keeps the stoichiometry of the two Microprocessor components in balance and thereby maintains consistent output of the miRNA biogenesis pathway; microarray analysis further suggests that the Drosha-DGCR8 complex acts on a broader range of hairpin-containing mRNAs beyond DGCR8's own transcript, extending Microprocessor's regulatory reach to additional gene targets in a manner independent of downstream miRNA-mediated silencing.

Specificity

Drosha Antibody (Rabbit mAb) [M15H18] detects endogenous levels of total Drosha proteins.

Background

Drosha is the catalytic ribonuclease III enzyme of the Microprocessor complex, functioning together with its obligate RNA-binding partner DGCR8 to catalyze the first, nuclear step of microRNA biogenesis: cleavage of the hairpin structures embedded within long primary microRNA transcripts (pri-miRNAs) to release the shorter precursor miRNA (pre-miRNA) that is subsequently exported to the cytoplasm for further processing by Dicer. Cryo-electron microscopy structures of the Drosha-DGCR8 complex, solved both with and without a bound pri-miRNA, show that a helix-hairpin element within an extended PAZ domain of Drosha, together with a mobile basic helix in Drosha's RNase IIIa domain, jointly recognize the single-stranded-to-double-stranded junction of the pri-miRNA hairpin, while Drosha's double-stranded RNA-binding domain makes extensive contacts along the RNA stem; the RNA-free structure additionally reveals that this same PAZ helix-hairpin element adopts an autoinhibitory conformation in the absence of substrate, indicating that pri-miRNA binding itself triggers a conformational change required to position Drosha's catalytic centers for correct, sequence-independent cleavage site selection. DGCR8 is required not only for RNA substrate recognition through its own double-stranded RNA-binding domains but also for stabilizing Drosha itself: DGCR8 positively regulates Drosha protein levels through a direct protein-protein interaction, while Drosha reciprocally regulates DGCR8 by cleaving conserved hairpin structures embedded within the DGCR8 mRNA itself, a Microprocessor-mediated cleavage event that destabilizes the DGCR8 transcript and reduces DGCR8 protein output. This reciprocal, cross-regulatory relationship, in which Drosha stabilizes its partner at the protein level while DGCR8 destabilizes Drosha's partner at the mRNA level, establishes a homeostatic feedback loop that keeps the stoichiometry of the two Microprocessor components in balance and thereby maintains consistent output of the miRNA biogenesis pathway; microarray analysis further suggests that the Drosha-DGCR8 complex acts on a broader range of hairpin-containing mRNAs beyond DGCR8's own transcript, extending Microprocessor's regulatory reach to additional gene targets in a manner independent of downstream miRNA-mediated silencing.

使用情報

Application

WB, IP

Dilution

WB	IP
1:1000	1:50

Reactivity

Human, Mouse

Source

Rabbit Monoclonal Antibody

MW

159 kDa

Storage Buffer

PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage
(from the date of receipt)

-20°C (avoid freeze-thaw cycles), 2 years

プロトコール

References

https://pubmed.ncbi.nlm.nih.gov/19135890/
https://pubmed.ncbi.nlm.nih.gov/32220645/

Drosha Antibody (Rabbit mAb) [M15H18]

生物学的記述

使用情報

References

Application Data