Identifying and Validating MYC:Protein Interactors in Pursuit of Novel Anti-MYC Therapies

Area of Science:

• Molecular Biology
• Oncology
• Biochemistry

Background:

• The MYC oncoprotein is a critical regulator of gene transcription and a driver of tumorigenesis.
• Understanding MYC's protein-protein interactions is essential for elucidating its oncogenic mechanisms.
• Targeting MYC interactions offers a potential therapeutic strategy for MYC-driven cancers.

Purpose of the Study:

• To detail methodologies for identifying and validating MYC protein-protein interactors.
• To provide a framework for understanding MYC's role in gene regulation and cancer.
• To enable the development of strategies that disrupt MYC interactions for cancer therapy.

Main Methods:

• BioID: Proximity-dependent biotinylation for identifying proximal interactors in living cells.
• Bioinformatic prioritization: Strategies for analyzing BioID-derived MYC-proximal complexes.
• Validation techniques: In vivo-in vitro pull-down assays and proximity ligation assay (PLA).
• Quantitative analysis: Biolayer interferometry (BLI) for characterizing direct protein interactions in vitro.

Main Results:

• The chapter outlines four distinct techniques for MYC interactor discovery and validation.
• Methodologies cover both in vivo proximity labeling and in vitro interaction characterization.
• The presented methods are adaptable for studying interactors of other proteins of interest.

Conclusions:

• Comprehensive identification and validation of MYC interactors are crucial for understanding its function.
• The discussed techniques provide a robust toolkit for interrogating MYC's molecular network.
• Disrupting key MYC interactions holds promise for inhibiting MYC-driven tumorigenesis.