TimeLapse-seq reveals that ALKBH5 modulates RNA stability of metabolic transcripts. TimeLapse-seq reveals that ALKBH5 modulates RNA stability of metabolic transcripts

N6-methyadenosine (m6A) RNA modification controls numerous cellular processes through regulation of RNA stability and translational efficiency. Whether the RNA m6A modification regulates energy metabolism process by posttranscriptional regulation is unknown. We here show that loss of the m6A demethylase ALKBH5 results in instability of oxogluatarate-dehydrogenase (Ogdh) messenger RNA and transcripts of other metabolic enzymes. Overall design: RNA isolated from wildtype and Alkbh5-deficient murine lineage-depleted bone marrow cells treated +/- s4U was subjected to oxidative-nucleophilic-aromatic-substitution chemistry. Sequencing libraries were prepared from all RNA.

N6-甲基腺嘌呤（N6-methyadenosine，m6A）RNA修饰通过调控RNA稳定性与翻译效率，调控诸多细胞生命活动。目前尚不明确RNA m6A修饰是否可通过转录后调控参与能量代谢过程。本研究证实，m6A去甲基化酶ALKBH5的缺失会导致氧戊二酸脱氢酶（oxogluatarate-dehydrogenase，Ogdh）的信使RNA以及其他代谢酶的转录本稳定性降低。实验设计概述：从野生型及Alkbh5缺陷型小鼠的谱系剔除骨髓细胞中提取RNA，将这些细胞分别经4-硫代尿苷（s4U）处理与未经处理，随后对提取的RNA实施氧化-亲核芳香取代化学反应，并基于所有上述RNA样品构建测序文库。