Institut de Biologie de l'École Normale Supérieure (IBENS), Département de Biologie, Ecole Normale Supérieure, CNRS, Inserm, PSL Research University, F-75005 Paris, France.
Physics Department, East China Normal University, Shanghai, China.
Nucleic Acids Res. 2018 Nov 2;46(19):10157-10172. doi: 10.1093/nar/gky797.
The spatiotemporal program of metazoan DNA replication is regulated during development and altered in cancers. We have generated novel OK-seq, Repli-seq and RNA-seq data to compare the DNA replication and gene expression programs of twelve cancer and non-cancer human cell types. Changes in replication fork directionality (RFD) determined by OK-seq are widespread but more frequent within GC-poor isochores and largely disconnected from transcription changes. Cancer cell RFD profiles cluster with non-cancer cells of similar developmental origin but not with different cancer types. Importantly, recurrent RFD changes are detected in specific tumour progression pathways. Using a model for establishment and early progression of chronic myeloid leukemia (CML), we identify 1027 replication initiation zones (IZs) that progressively change efficiency during long-term expression of the BCR-ABL1 oncogene, being twice more often downregulated than upregulated. Prolonged expression of BCR-ABL1 results in targeting of new IZs and accentuation of previous efficiency changes. Targeted IZs are predominantly located in GC-poor, late replicating gene deserts and frequently silenced in late CML. Prolonged expression of BCR-ABL1 results in massive deletion of GC-poor, late replicating DNA sequences enriched in origin silencing events. We conclude that BCR-ABL1 expression progressively affects replication and stability of GC-poor, late-replicating regions during CML progression.
真核生物 DNA 复制的时空程序在发育过程中受到调控,并在癌症中发生改变。我们生成了新的 OK-seq、Repli-seq 和 RNA-seq 数据,以比较 12 种人类癌细胞和非癌细胞的 DNA 复制和基因表达程序。OK-seq 确定的复制叉方向(RFD)变化广泛,但在 GC 贫乏的同型异序区更为频繁,并且与转录变化基本没有关联。癌细胞的 RFD 图谱与具有相似发育起源的非癌细胞聚类,但与不同的癌症类型不聚类。重要的是,在特定的肿瘤进展途径中检测到反复出现的 RFD 变化。使用慢性髓性白血病(CML)建立和早期进展的模型,我们鉴定了 1027 个复制起始区(IZ),在 BCR-ABL1 癌基因的长期表达过程中,其效率逐渐变化,下调的频率是上调的两倍。BCR-ABL1 的长期表达导致新的 IZ 靶向和先前效率变化的加重。靶向 IZ 主要位于 GC 贫乏、晚期复制的基因荒漠中,并且在晚期 CML 中经常沉默。BCR-ABL1 的长期表达导致富含起始沉默事件的 GC 贫乏、晚期复制 DNA 序列的大规模缺失。我们得出结论,BCR-ABL1 的表达在 CML 进展过程中逐渐影响 GC 贫乏、晚期复制区域的复制和稳定性。