E2F regulation of Phosphoglycerate kinase (Pgk) gene is functionally important in Drosophila development. E2F regulation of Phosphoglycerate kinase (Pgk) gene is functionally important in Drosophila development

Our study focuses on understanding the changes in chromatin accessibility in the Pgk-DeltaE2F mutant that takes place during flight muscles development in Drosophila. E2F binding sites were mutated upstream Pgk gene. E2F/Dp/Rbf were no longer recruited to this regulatory region. As a result of this, the expression of Pgk mRNA was downregulated, which led to reduced levels in many metabolites that are glycolytic and TCA intermediates. Using ATAC-seq we determined that there was a global reduction in chromatin accessibility in the mutant compared to control, and it was accompagnied by a widespread reduction in gene expression, including metabolic genes. In addition we identified a broad range of phenotypes when analyzing the functional relevance of E2F regulation on Pgk gene. These phenotypes range from low ATP content, abnormal mitochondrial morphology, shortened life span to embrionic lethality. Furthermore, defects in high-energy consuming organs, such as ovaries and muscles, were found. In sum, our results illustrate the pleiotropic effects on metabolism, gene expression and development in the Pgk-deltaE2F animals, which underlies the importance of E2F regulation on a single E2F target, Pgk. Overall design: ATAC-Seq on nuclei extracted from thoracic muscles enriched for flight muscles staged at pharate (96h APF). The Pgk.deltaE2F mutant was compared to control. A total of 2 replicates per genotype were processed.

本研究旨在解析果蝇（Drosophila）飞行肌发育过程中，Pgk-DeltaE2F突变体的染色质可及性（chromatin accessibility）变化。研究中对Pgk基因上游的E2F结合位点进行了突变修饰，导致E2F/Dp/Rbf复合物无法再被招募至该调控区域。受此影响，Pgk基因的mRNA表达水平下调，进而使得多种糖酵解及三羧酸（TCA）循环中间代谢物的含量显著降低。我们通过转座酶可及性染色质测序（ATAC-seq）检测发现，相较于野生型对照，突变体的全基因组染色质可及性整体降低，且伴随包括代谢基因在内的大量基因的表达水平广泛下调。此外，在分析E2F对Pgk基因的调控功能相关性时，我们观测到一系列表型变化：这些表型涵盖ATP含量降低、线粒体形态异常、寿命缩短乃至胚胎致死；同时还发现卵巢、肌肉等高能量消耗器官存在功能缺陷。综上，本研究结果揭示了Pgk-deltaE2F突变体在代谢、基因表达及发育层面的多效性影响，阐明了单个E2F靶基因Pgk的E2F调控的重要性。实验设计：对处于预蛹期（96h APF）、富集有飞行肌的胸部肌肉组织提取的细胞核进行ATAC-seq测序，以Pgk.deltaE2F突变体与野生型对照进行对比，每种基因型设置2个生物学重复。