Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore 560064, India.
National laboratory of Bio-macromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing 100101, China.
Nucleic Acids Res. 2022 Jun 24;50(11):6116-6136. doi: 10.1093/nar/gkac450.
Human Positive Coactivator 4 (PC4) is a multifaceted chromatin protein involved in diverse cellular processes including genome organization, transcription regulation, replication, DNA repair and autophagy. PC4 exists as a phospho-protein in cells which impinges on its acetylation by p300 and thereby affects its transcriptional co-activator functions via double-stranded DNA binding. Despite the inhibitory effects, the abundance of phosphorylated PC4 in cells intrigued us to investigate its role in chromatin functions in a basal state of the cell. We found that casein kinase-II (CKII)-mediated phosphorylation of PC4 is critical for its interaction with linker histone H1. By employing analytical ultracentrifugation and electron microscopy imaging of in vitro reconstituted nucleosomal array, we observed that phospho-mimic (PM) PC4 displays a superior chromatin condensation potential in conjunction with linker histone H1. ATAC-sequencing further unveiled the role of PC4 phosphorylation to be critical in inducing chromatin compaction of a wide array of coding and non-coding genes in vivo. Concordantly, phospho-PC4 mediated changes in chromatin accessibility led to gene repression and affected global histone modifications. We propose that the abundance of PC4 in its phosphorylated state contributes to genome compaction contrary to its co-activator function in driving several cellular processes like gene transcription and autophagy.
人类阳性共激活因子 4(PC4)是一种多功能染色质蛋白,参与多种细胞过程,包括基因组组织、转录调控、复制、DNA 修复和自噬。PC4 在细胞中作为一种磷酸化蛋白存在,其磷酸化会影响 p300 的乙酰化,从而通过双链 DNA 结合影响其转录共激活因子功能。尽管存在抑制作用,但细胞中磷酸化 PC4 的丰度引起了我们的兴趣,以研究其在细胞基础状态下染色质功能中的作用。我们发现,酪蛋白激酶-II(CKII)介导的 PC4 磷酸化对于其与连接组蛋白 H1 的相互作用至关重要。通过采用分析超速离心和体外重建核小体阵列的电子显微镜成像,我们观察到磷酸模拟(PM)PC4 与连接组蛋白 H1 一起显示出优越的染色质浓缩潜力。ATAC 测序进一步揭示了 PC4 磷酸化在体内诱导广泛的编码和非编码基因染色质紧缩中的关键作用。一致地,磷酸化 PC4 介导的染色质可及性变化导致基因抑制,并影响全局组蛋白修饰。我们提出,PC4 在其磷酸化状态下的丰度有助于基因组紧缩,与它在驱动基因转录和自噬等多种细胞过程中的共激活因子功能相反。