Identification of MYC intron 2 regions that modulate expression

Area of Science:

• Molecular Biology
• Gene Regulation
• RNA Splicing

Background:

• MYC transcription factor is vital for cellular processes like differentiation and proliferation.
• The precise mechanisms governing MYC pre-mRNA splicing are not well understood.
• Heterogeneous nuclear ribonucleoprotein C (HNRNPC) is a key RNA-binding protein.

Purpose of the Study:

• To investigate the interaction between HNRNPC and MYC intron 2.
• To elucidate the role of HNRNPC in MYC pre-mRNA splicing and translation.
• To identify regulatory elements within MYC intron 2.

Main Methods:

• Utilized published eCLIP data to confirm HNRNPC binding sites.
• Performed mutational analysis of poly(U) regions in MYC intron 2.
• Analyzed effects on HNRNPC binding, splicing fidelity, and translation efficiency.
• Examined sequence conservation of identified binding regions across species.

Main Results:

• Confirmed HNRNPC binds to specific poly(U) regions within MYC intron 2.
• Demonstrated that disruption of all three poly(U) sites significantly reduces HNRNPC binding and compromises splicing and translation fidelity.
• Found that these poly(U) sequences are conserved across multiple species.
• Identified a novel regulatory element upstream of an HNRNPC binding site that enhances MYC translation when deleted.

Conclusions:

• HNRNPC plays a critical, compensatory role in maintaining MYC pre-mRNA splicing and translation fidelity through its interaction with intronic poly(U) regions.
• The identified conserved sequences and regulatory elements offer new targets for understanding and potentially manipulating MYC gene expression.