DPF2 reads histone lactylation to drive transcription and tumorigenesis [CUT&TAG]

Lysine lactylation (Kla) is a new type of histone mark implicated in the regulation of various functional processes such as transcription. However, how this histone mark acts in cancers remains unexplored due in part to a lack of knowledge about its reader proteins. Here, we observe that cervical cancer (CC) cells undergo metabolic reprogram by which lactate accumulation and thereby boost histone lactylation, particularly H3K14la. Utilizing a multivalent photoaffinity probe in combination with quantitative proteomics approach, we identify DPF2 as a candidate target of H3K14la. Biochemical studies as well as CUT&Tag analysis reveal that DPF2 is capable of binding to H3K14la, and co-localizes with it on promoters of oncogenic genes. Notably, disrupting the association between DPF2 and histone lactylation through structure-guided mutation blunts those cancer-related gene expression along with cell survival. Together, our findings reveal DPF2 as a bona fide H3K14la effector that couples histone lactylation to gene transcription and cell survival, offering insight into how histone Kla engages in transcription and tumorigenesis. We implemented CUT&Tag assays in Hela cells to test whether DPF2 links H3K14la in the native chromatin context. To this end, the cells were immunoprecipitated by H3K14la or H3K14ac antibodies, respectively, followed by high-throughput sequencing. Furthermore, to test whether DPF2-occupied peaks are enriched in a DPF-domain-dependent manner, cells stably expressing Flag-tagged DPF2 or its mutants were immunoprecipitated using Flag antibodies and analyzed by sequencing.