Sinha Kalyan, Bilokapic Silvija, Du Yongming, Malik Deepshikha, Halic Mario
Department of Structural Biology, St. Jude Children's Research Hospital, 262 Danny Thomas Place, Memphis, TN, 38105, USA.
bioRxiv. 2023 Mar 15:2023.03.14.532583. doi: 10.1101/2023.03.14.532583.
Pioneer transcription factors have the ability to access DNA in compacted chromatin. Multiple transcription factors can bind together to a regulatory element in a cooperative way and cooperation between pioneer transcription factors Oct4 and Sox2 is important for pluripotency and reprogramming. However, the molecular mechanisms by which pioneer transcription factors function and cooperate remain unclear. Here we present cryo-EM structures of human Oct4 bound to a nucleosome containing human Lin28B and nMatn1 DNA sequences, which bear multiple binding sites for Oct4. Our structural and biochemistry data reveal that Oct4 binding induces changes to the nucleosome structure, repositions the nucleosomal DNA and facilitates cooperative binding of additional Oct4 and of Sox2 to their internal binding sites. The flexible activation domain of Oct4 contacts the histone H4 N-terminal tail, altering its conformation and thus promoting chromatin decompaction. Moreover, the DNA binding domain of Oct4 engages with histone H3 N-terminal tail, and posttranslational modifications at H3K27 modulate DNA positioning and affect transcription factor cooperativity. Thus, our data show that the epigenetic landscape can regulate Oct4 activity to ensure proper cell reprogramming.
先驱转录因子能够在紧密的染色质中接触到DNA。多种转录因子可以以协同的方式共同结合到一个调控元件上,而先驱转录因子Oct4和Sox2之间的协同作用对于多能性和重编程至关重要。然而,先驱转录因子发挥功能和协同作用的分子机制仍不清楚。在此,我们展示了与包含人类Lin28B和nMatn1 DNA序列的核小体结合的人类Oct4的冷冻电镜结构,这些序列带有多个Oct4的结合位点。我们的结构和生化数据表明,Oct4的结合会诱导核小体结构发生变化,重新定位核小体DNA,并促进额外的Oct4和Sox2与其内部结合位点的协同结合。Oct4灵活的激活结构域与组蛋白H4的N端尾巴接触,改变其构象,从而促进染色质解压缩。此外,Oct4的DNA结合结构域与组蛋白H3的N端尾巴相互作用,H3K27处的翻译后修饰调节DNA定位并影响转录因子的协同作用。因此,我们的数据表明表观遗传格局可以调节Oct4的活性,以确保正确的细胞重编程。