Energy status orchestrates YTHDF1 phase separation and tumorigenesis-Accepted Version

Aberrant energy status impacts the initiation and progression of tumorigenesis, although the underlying mechanisms remain poorly understood. Adenosine monophosphate (AMP)-activated protein kinase (AMPK), a key sensor of cellular energy stress, is activated to facilitate metabolic adaptation and regulate tumorigenesis. Here, we reveal that energy deprivation-induced activation of AMPK phosphorylates YTHDF1 at Ser198, counteracting its O-GlcNAcylation. This phosphorylation alters the functional properties of YTHDF1 by suppressing its phase separation and its interaction with the translation initiation factor eIF3b, ultimately reducing protein translation. Notably, enhancing YTHDF1 phosphorylation to antagonize its O-GlcNAcylation through AMPK agonists or ketogenic diet effectively inhibits tumor cell growth both in vitro and in vivo. These findings elucidate a regulatory mechanism that links cellular energy status to YTHDF1 post-translational modifications and highlight the therapeutic potential of targeting YTHDF1-mediated pathways via metabolic interventions for cancer treatment.异常的能量状态会影响肿瘤发生的起始与进展，但其潜在分子机制仍不甚明确。腺苷一磷酸（AMP）激活的蛋白激酶（AMPK）作为细胞能量应激的关键感受器，可被激活以促进代谢适应并调控肿瘤发生。本研究发现，能量剥夺诱导的AMPK激活会在丝氨酸198（Ser198）位点磷酸化YTHDF1，从而拮抗其O-连接β-N-乙酰葡糖胺糖基化（O-GlcNAcylation）修饰。该磷酸化事件通过抑制YTHDF1的相分离能力及其与翻译起始因子eIF3b的相互作用，改变其功能特性，最终减少蛋白质翻译过程。值得注意的是，通过AMPK激动剂或生酮饮食增强YTHDF1的磷酸化以拮抗其O-连接β-N-乙酰葡糖胺糖基化修饰，可在体外与体内实验中有效抑制肿瘤细胞的生长。本研究揭示了一条将细胞能量状态与YTHDF1翻译后修饰相联系的调控机制，并凸显了通过代谢干预靶向YTHDF1介导的通路以治疗癌症的治疗潜力。