Wee1 opposes APC/C(Cdh1) activity to promote S-phase entry

Vuillemenot, L.-A.; Morales, L. G.; Dragoi, C.-M.; Pennycook, B. R.; Harris, E.; Lin, L. C.; Weston, W. A.; Brown, J.-E.; Graham, E.; Andrew, I.; Roumelioti, G.; Montoya, A.; Game, L.; Young, G. R.; Shliaha, P.; Novak, B.; Barr, A. R.

doi:10.64898/2025.12.01.691493

bioRxiv · 10.64898/2025.12.01.691493

Wee1 opposes APC/C(Cdh1) activity to promote S-phase entry

Vuillemenot, L.-A.·Morales, L. G.·Dragoi, C.-M.·Pennycook, B. R.·Harris, E.·Lin, L. C.·Weston, W. A.·Brown, J.-E.·Graham, E.·Andrew, I.·Roumelioti, G.·Montoya, A.·Game, L.·Young, G. R.·Shliaha, P.·Novak, B.·Barr, A. R.

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Abstract

Wee1 phosphorylates and inhibits CDK activity to inhibit mitotic entry and establish a G2 DNA damage checkpoint. Consequently, Wee1 inhibitors are in clinical trials, developed to be synthetically lethal in TP53 mutant tumours that become reliant on a Wee1-mediated DNA damage checkpoint. However, Wee1 inhibitors have efficacy in TP53 wild-type tumours and many trials have been terminated due to high levels of toxic side-effects, suggesting that Wee1 has unknown functions. Here, we show that Wee1 promotes cell cycle re-entry from quiescence (G0) by opposing the activity of the E3 ubiquitin ligase, APC/CCdh1. Wee1 phosphorylates Cdh1 (FZR1) at key residues that mediate the interaction between Cdh1 and APC/C. Cells with loss-of-function of Wee1 during G0/G1 have delayed S-phase entry, an impaired G1/S transition, abnormal S-phase accumulation of the CDK inhibitor p21 and enter a p21-dependent G2 arrest. Reduced expression of APC/CCdh1 or p21 renders cells more sensitive to acute Wee1 inhibition and both pathways are downregulated in acquired Wee1 inhibitor resistance. Our study reveals a new cell cycle control mechanism that has implications for how Wee1 inhibitors should be used in the clinic.

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Vuillemenot, L.-A., Morales, L. G., Dragoi, C.-M., Pennycook, B. R., Harris, E., Lin, L. C., Weston, W. A., Brown, J.-E., Graham, E., Andrew, I., Roumelioti, G., Montoya, A., Game, L., Young, G. R., Shliaha, P., Novak, B., Barr, A. R.. 2025-12-02. Wee1 opposes APC/C(Cdh1) activity to promote S-phase entry. https://doi.org/10.64898/2025.12.01.691493

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