School of Life Sciences, Jilin University, Changchun 130012, China.
School of Pharmacy, Changchun University of Chinese Medicine, Changchun 130117, China.
Int J Mol Sci. 2022 Mar 30;23(7):3801. doi: 10.3390/ijms23073801.
The human males absent on the first (MOF)-containing non-specific lethal (NSL) histone acetyltransferase (HAT) complex acetylates histone H4 at lysine K5, K8, and K16. This complex shares several subunits with other epigenetic regulatory enzymes, which highlights the complexity of its intracellular function. However, the effect of the NSL HAT complex on the genome and target genes in human cells is still unclear. By using a CRISPR/Cas9-mediated NSL3-knockout 293T cell line and chromatin immunoprecipitation-sequencing (ChIP-Seq) approaches, we identified more than 100 genes as NSL HAT transcriptional targets, including several transcription factors, such as () which are mainly involved in cell proliferation, biological adhesion, and metabolic processes. We found here that the ChIP-Seq peaks of MOF and NSL3 co-localized with H4K16ac, H3K4me2, and H3K4me3 at the transcriptional start site of YY1. In addition, both the mRNA and protein expression levels of YY1 were regulated by silencing or overexpressing NSL HAT. Interestingly, the expression levels of cell division cycle 6, a downstream target gene of YY1, were regulated by MOF or NSL3. In addition, the suppressed clonogenic ability of HepG2 cells caused by siNSL3 was reversed by overexpressing YY1, suggesting the involvement of YY1 in NSL HAT functioning. Additionally, de novo motif analysis of MOF and NSL3 targets indicated that the NSL HAT complex may recognize the specific DNA-binding sites in the promoter region of target genes in order to regulate their transcription.
人类雄性个体在包含第一个（MOF）的非特异性致死（NSL）组蛋白乙酰转移酶（HAT）复合物中不存在，该复合物在赖氨酸 K5、K8 和 K16 处乙酰化组蛋白 H4。该复合物与其他表观遗传调节酶共享几个亚基，这突出了其细胞内功能的复杂性。然而，NSL HAT 复合物对人类细胞基因组和靶基因的影响仍不清楚。通过使用 CRISPR/Cas9 介导的 NSL3 敲除 293T 细胞系和染色质免疫沉淀测序（ChIP-Seq）方法，我们鉴定出超过 100 个基因作为 NSL HAT 的转录靶标，其中包括几个转录因子，如 ()，它们主要参与细胞增殖、生物粘附和代谢过程。我们发现这里 MOF 和 NSL3 的 ChIP-Seq 峰与 YY1 的转录起始位点处的 H4K16ac、H3K4me2 和 H3K4me3 共定位。此外，YY1 的 mRNA 和蛋白表达水平均受 NSL HAT 的沉默或过表达调节。有趣的是，YY1 的下游靶基因细胞分裂周期 6 的表达水平受 MOF 或 NSL3 调节。此外，siNSL3 抑制 HepG2 细胞的集落形成能力被过表达 YY1 逆转，表明 YY1 参与 NSL HAT 功能。此外，MOF 和 NSL3 靶标的从头 motif 分析表明，NSL HAT 复合物可能识别靶基因启动子区域中的特定 DNA 结合位点，以调节其转录。
