YAP engages RIF1 to dampen replication stress-induced DNA damage in human squamous cell carcinoma

Squamous cell carcinoma cells experience high levels of replication stress due to oncogene-induced cell cycle deregulation. How such cells sustain rapid proliferation despite replication stress is still not fully understood. Here, we discovered, using rapid immunoprecipitation mass spectrometry of endogenous protein (RIME) analysis, that the squamous cell carcinoma oncoprotein YAP engages with RIF1, a key regulator of DNA replication and DNA damage repair under replication stress. RIF1 is highly expressed in human squamous cell carcinoma cell lines and tissues and upregulated during tumour progression. Depletion of RIF1 in squamous cell carcinoma cells exacerbates their endogenous replication stress. Mechanistically, we show that YAP interacts with RIF1 specifically at broken replication forks, and that YAP depletion impairs DNA damage repair under replication stress. Our results thus demonstrate that YAP’s oncogenic functions in squamous cancers involve both transcriptional and non-transcriptional mechanisms, the latter through interaction with RIF1 to dampen replication stress.