Authors:

Sridhar Thiru

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About Paper:

Squamous Cell Carcinoma (SCC) is an aggressive malignancy driven in part by dysregulated protein synthesis. Cancer cells rewire translation to upregulate oncogenic proteins, making translational regulators attractive therapeutic targets. elF2A (eukaryotic initiation factor 2A) is highly expressed in SCC, lung, and ovarian cancers and correlates with poor patient prognosis, yet the lack of structural and functional characterization has impeded drug development efforts.'? We hypothesize that elF2A promotes tumorigenesis by remodeling translation of oncogenic transcripts, and that disrupting its interaction with the 40S ribosomal subunit will inhibit cancer cell growth. Using biochemical and structural approaches, we have characterized the elF2A-40S interaction. elF2A comprises three main domains: C-terminus a-helical domains (RAD1 and RAD2) and a B-propeller WD40 domain. Biolayer interferometry (BLI) revealed that full-length His-tagged elF2A binds 40S ribosomal subunits with a robust nanomolar affinity (Kd = 1.4 nM). Single-particle cryo-electron microscopy resolved the elF2A-40S complex to precise 2.8 Angstrom units, showing the WD40 domain positioned above the P-site while the C-terminal helical domains anchor to the 40S body. Mutational analyses confirmed that the C-terminal domain (residues 428-585) is responsible for 40S binding (Kd = 2.3 nM), while the isolated WD40 domain shows no detectable interaction. Building on this structural foundation, structure-guided small molecule inhibitors targeting the C-terminal helical domains of elF2A are being developed to disrupt 40S engagement. De novo virtual screening and Al-driven modeling using Boltz Gen, BindCraft, AlphaFold2, and ChimeraxX will be performed against commercially available compound libraries, with top candidates validated by BLI and cell-based assays in SCC cell lines. These studies aim to establish elF2A inhibition as a first-in-class therapeutic strategy for SCC and potentially other cancers driven by translational remodeling.

Source:

Northwestern University

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Co-authors:

Sridhar Thiru