Lee Kwangwoon, Barone Marco, Waterbury Amanda L, Jiang Hanjie, Nam Eunju, DuBois-Coyne Sarah E, Whedon Samuel D, Wang Zhipeng A, Caroli Jonatan, Neal Katherine, Ibeabuchi Brian, Dhoondia Zuzer, Kuroda Mitzi I, Liau Brian B, Beck Samuel, Mattevi Andrea, Cole Philip A
Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Boston, MA, USA.
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA.
Nat Chem Biol. 2025 Feb;21(2):227-237. doi: 10.1038/s41589-024-01671-9. Epub 2024 Jul 4.
Biochemical crosstalk between two or more histone modifications is often observed in epigenetic enzyme regulation, but its functional significance in cells has been difficult to discern. Previous enzymatic studies revealed that Lys14 acetylation of histone H3 can inhibit Lys4 demethylation by lysine-specific demethylase 1 (LSD1). In the present study, we engineered a mutant form of LSD1, Y391K, which renders the nucleosome demethylase activity of LSD1 insensitive to Lys14 acetylation. K562 cells with the Y391K LSD1 CRISPR knockin show decreased expression of a set of genes associated with cellular adhesion and myeloid leukocyte activation. Chromatin profiling revealed that the cis-regulatory regions of these silenced genes display a higher level of H3 Lys14 acetylation, and edited K562 cells show diminished H3 mono-methyl Lys4 near these silenced genes, consistent with a role for enhanced LSD1 demethylase activity. These findings illuminate the functional consequences of disconnecting histone modification crosstalk for a key epigenetic enzyme.
在表观遗传酶调控中，经常观察到两种或更多种组蛋白修饰之间的生化串扰，但其在细胞中的功能意义一直难以辨别。先前的酶学研究表明，组蛋白H3的赖氨酸14乙酰化可抑制赖氨酸特异性去甲基化酶1（LSD1）介导的赖氨酸4去甲基化。在本研究中，我们构建了一种LSD1的突变形式Y391K，它使LSD1的核小体去甲基化酶活性对赖氨酸14乙酰化不敏感。携带Y391K LSD1 CRISPR敲入的K562细胞显示，一组与细胞黏附和髓系白细胞活化相关的基因表达降低。染色质分析表明，这些沉默基因的顺式调控区域显示出较高水平的H3赖氨酸14乙酰化，并且编辑后的K562细胞在这些沉默基因附近显示出H3单甲基赖氨酸4减少，这与LSD1去甲基化酶活性增强的作用一致。这些发现阐明了切断关键表观遗传酶的组蛋白修饰串扰的功能后果。
