DPF2 reads histone lactylation to drive transcription and tumorigenesis [RNA-seq]. DPF2 reads histone lactylation to drive transcription and tumorigenesis [RNA-seq]

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. Overall design: On the basis of our findings that the correlation between DPF2 and H3K14la at promoter regions and lactate boosts H3K14la instead of H3K14ac, we propose that DPF2-H3K14la recognition confer transcription, and the genes modulated by this recognition could be obtained through a quantitative RNA-Seq analysis of cells expressing WT-DPF2 and Mut-DPF2 prior to lactate supplementation. To assess this, we performed RNA-Seq detection from WT-DPF2 or Mut-DPF2 expressed cells upon lactate pre-treatment.

赖氨酸乳酰化（Kla，Lysine lactylation）是一类新型组蛋白修饰，参与转录等多种生物学功能过程的调控。然而，由于对其组蛋白读取蛋白（reader proteins）的认知不足，这类组蛋白修饰在癌症中的作用机制仍未被阐明。本研究中，我们观察到宫颈癌细胞（CC，cervical cancer）会发生代谢重编程，通过该过程实现乳酸堆积，进而促进组蛋白乳酰化，尤其是H3K14la修饰。我们利用多价光亲和探针结合定量蛋白质组学方法，将DPF2鉴定为H3K14la的候选结合蛋白。生化实验与CUT&Tag分析结果表明，DPF2能够特异性结合H3K14la，并在致癌基因的启动子区域与该修饰共定位。值得注意的是，通过结构指导的突变破坏DPF2与组蛋白乳酰化的结合，会削弱相关致癌基因的表达水平并抑制细胞存活。综上，本研究证实DPF2是确凿的H3K14la效应蛋白，可将组蛋白乳酰化与基因转录及细胞存活过程相耦联，为解析组蛋白乳酰化如何参与转录调控与肿瘤发生提供了全新视角。

整体实验设计：基于我们的前期发现——启动子区域DPF2与H3K14la存在共定位关联，且乳酸可特异性促进H3K14la修饰而非H3K14乙酰化修饰——我们提出假设：DPF2与H3K14la的识别作用可调控基因转录，而该识别通路所调控的基因可通过对乳酸预处理前分别表达野生型DPF2（WT-DPF2）与突变型DPF2（Mut-DPF2）的细胞进行定量RNA测序（RNA-Seq）分析获得。为验证该假设，我们对乳酸预处理后的表达WT-DPF2或Mut-DPF2的细胞开展了RNA测序检测。