CENP-A associated lncRNAs influence chromosome segregation in human cells. CENP-A associated lncRNAs influence chromosome segregation in human cells

Transcription occurs ubiquitously throughout non-coding parts of the genome, including at repetitive alpha-satellite DNA elements which comprise the majority of human centromeres. The function of temporally regulated centromeric transcription, and transcripts, is consequently a topic of intense investigation. In this study, we use high throughput approaches to identify and describe lncRNAs associated with the centromere specific histone variant CENP-A that arise from the transcription of specific centromeres at early G1, which we then show are physically associated with centromeres, and which are functionally necessary for accurate chromosome segregation. Targeted depletion of one such centromeric RNA, which originates from a single centromere, is sufficient to increase the frequency of chromosome segregation defects. These data support the emerging paradigm of the necessity of centromere-specific lncRNAs in the integrity of faithful chromosome segregation. Overall design: RNA-immunoprecipitation followed by Illumina sequencing (RIP-Seq) on chromatin associated or nucleoplasmic RNA.

转录广泛分布于基因组的非编码区域，包括构成人类着丝粒主体的重复α-卫星DNA（alpha-satellite DNA）元件。因此，受时间调控的着丝粒转录及其转录本的功能，是当前广受关注的研究课题。本研究采用高通量实验手段，对细胞周期早期G1阶段由特定着丝粒转录产生的、与着丝粒特异性组蛋白变体CENP-A结合的长链非编码RNA（long non-coding RNAs，lncRNAs）进行鉴定与表征；后续实验证实，此类RNA可物理结合于着丝粒，且对精准染色体分离具有功能必要性。靶向耗竭源自单个着丝粒的此类着丝粒RNA之一，即可显著提升染色体分离缺陷的发生频率。本研究数据支持了新兴的学术范式：着丝粒特异性长链非编码RNA对于维持染色体精准分离的完整性不可或缺。实验整体设计：对染色质结合RNA或核质RNA实施RNA免疫共沉淀（RNA-immunoprecipitation）后，开展Illumina测序（RIP-Seq）。