DBP10 Antibody

Introduction to DBP10 Antibody

The DBP10 antibody is a research tool designed to target the yeast DEAD-box RNA helicase Dbp10, a critical protein involved in ribosome biogenesis. Dbp10 facilitates the maturation of 60S ribosomal subunits by restructuring pre-rRNA and coordinating the integration of assembly factors . Below is a detailed analysis of its function, research significance, and experimental applications.

Functional Role of Dbp10

Dbp10 operates in the nucleolus, where it binds pre-60S ribosomal particles and regulates RNA conformational changes essential for peptidyl-transferase center (PTC) formation . Key findings include:

• Catalytic Activity: Mutations in Dbp10’s ATP-binding/hydrolysis motifs (Walker A/B) impair rRNA methylation and delay pre-60S maturation .

• Interactions: Dbp10 associates with GTPase Nug1 and methyltransferase Spb1, enabling 25S rRNA modification at nucleotide G2922 .

• Phenotypic Consequences: Dominant-negative mutations in Dbp10 lead to a lethal growth phenotype and accumulation of immature rRNA intermediates .

Research Applications of DBP10 Antibody

The antibody is used to study Dbp10’s localization, interaction networks, and functional dynamics. Common techniques include:

• Immunoprecipitation: To isolate pre-60S particles and analyze cofactors (e.g., Nug1, Spb1) .

• Western Blotting: Detection of Dbp10 in nucleolar fractions or mutant strains .

• Immunofluorescence: Visualizing Dbp10’s colocalization with nucleolar markers (e.g., fibrillarin) .

4.1. Helicase Activity

Mutational studies (K187R, E289A) reveal that Dbp10’s ATPase activity is critical for Nug1 recruitment and rRNA methylation . Antibody-based pulldowns confirmed reduced Spb1 incorporation in mutant particles .

4.2. rRNA Methylation

Dbp10 facilitates 2′-O-methylation of G2922 by Spb1, as shown by reverse transcription stops in wild-type vs. K187R mutant extracts .

4.3. Genetic Depletion

Cold-sensitive alleles (e.g., dbp10-ts) cause 60S subunit deficits and half-mer polysome accumulation, detectable via sucrose gradient analysis .