Bio@SNS Laboratory, Scuola Normale Superiore, 56126 Pisa, Italy.
European Molecular Biology Laboratory, Genome Biology Unit, 69117 Heidelberg, Germany.
Int J Mol Sci. 2022 Aug 10;23(16):8892. doi: 10.3390/ijms23168892.
Among Histone post-translational modifications (PTMs), lysine acetylation plays a pivotal role in the epigenetic regulation of gene expression, mediated by chromatin modifying enzymes. Due to their activity in physiology and pathology, several chemical compounds have been developed to inhibit the function of these proteins. However, the pleiotropy of these classes of proteins represents a weakness of epigenetic drugs. Ideally, a new generation of epigenetic drugs should target with molecular precision individual acetylated lysines on the target protein. We exploit a PTM-directed interference, based on an intrabody (scFv-58F) that selectively binds acetylated lysine 9 of histone H3 (H3K9ac), to test the hypothesis that targeting H3K9ac yields more specific effects than inhibiting the corresponding HAT enzyme that installs that PTM. In yeast scFv-58F modulates, gene expression in a more specific way, compared to two well-established HAT inhibitors. This PTM-specific interference modulated expression of genes involved in ribosome biogenesis and function. In mammalian cells, the scFv-58F induces exclusive changes in the H3K9ac-dependent expression of specific genes. These results suggest the H3K9ac-specific intrabody as the founder of a new class of molecules to directly target histone PTMs, inverting the paradigm from inhibiting the writer enzyme to acting on the PTM.
在组蛋白翻译后修饰 (PTMs) 中,赖氨酸乙酰化在由染色质修饰酶介导的基因表达的表观遗传调控中起着关键作用。由于它们在生理学和病理学中的活性,已经开发了几种化学化合物来抑制这些蛋白质的功能。然而,这些蛋白质类别的多效性是表观遗传药物的一个弱点。理想情况下,新一代的表观遗传药物应该以分子精度针对目标蛋白上的单个乙酰化赖氨酸。我们利用基于内体 (scFv-58F) 的 PTM 靶向干扰,该内体特异性结合组蛋白 H3 的乙酰化赖氨酸 9 (H3K9ac),以检验以下假设:靶向 H3K9ac 比抑制安装该 PTM 的相应 HAT 酶产生更特异的效果。与两种成熟的 HAT 抑制剂相比,在酵母中,scFv-58F 以更特异的方式调节基因表达。这种 PTM 特异性干扰调节了参与核糖体生物发生和功能的基因的表达。在哺乳动物细胞中,scFv-58F 诱导特定基因的 H3K9ac 依赖性表达的独特变化。这些结果表明 H3K9ac 特异性内体作为一类新分子的基础,可以直接针对组蛋白 PTM,将抑制写入酶的范式转变为针对 PTM 的作用。