DRBD3 regulates long non-coding RNA abundance and cryptic splice site selection in trypanosomes.

Area of Science:

• Molecular Biology
• Genetics
• Parasitology

Background:

• Trypanosomes utilize post-transcriptional regulation for gene expression control.
• DRBD3 is an RNA-binding protein involved in mRNA metabolism.
• DRBD3 associates with the long non-coding RNA (lncRNA) grumpy in Trypanosoma brucei.

Purpose of the Study:

• To investigate the role of DRBD3 in lncRNA metabolism in trypanosomes.
• To elucidate the impact of DRBD3 depletion on both lncRNA and protein-coding gene expression.
• To understand DRBD3's function in splicing regulation.

Main Methods:

• DRBD3 depletion via silencing.
• Analysis of lncRNA and mRNA expression profiles.
• Investigation of poly(A) polymerase PAP1 levels.
• Assessment of alternative splicing events.

Main Results:

• DRBD3 depletion upregulates a subset of lncRNAs, potentially via cryptic splice site activation.
• Reduced PAP1 levels correlate with DRBD3 silencing, affecting lncRNA expression indirectly.
• DRBD3 loss impacts protein-coding gene processing, causing alternative trans-splicing and protein truncation.
• DRBD3 regulates RBP20 transcript splicing and the balance between trans- and cis-splicing.

Conclusions:

• DRBD3 acts as a high-level regulator of gene expression in trypanosomes.
• DRBD3 influences the expression landscape of both coding and non-coding genes.
• DRBD3 is crucial for maintaining proper RNA processing and splicing fidelity in trypanosomes.