DDX47, MeCP2, and other functionally heterogeneous factors protect cells from harmful R loops.
DDX47, MeCP2, and other functionally heterogeneous factors protect cells from harmful R loops.
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DOI：
10.1016/j.celrep.2023.112148
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发表时间：
2023-03-28
期刊：
影响因子：
8.8
通讯作者：
中科院分区：
文献类型：
--
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非预定的R环可能是基因组不稳定的一个来源，这是癌细胞的一个标志。尽管靶向蛋白质组学方法和特定突变体的细胞分析已经发现了可能参与R环稳态的因素，但我们报告了基于激活诱导胞苷脱氨酶（AID）靶向R环的能力，对其耗损导致R环的因素进行了更开放的筛选。对覆盖3205个蛋白编码基因的小干扰rna （sirna）引起的γ - h2ax进行免疫荧光分析，鉴定出59个潜在候选基因，其中13个被进一步分析，显示R环显著增加。这些候选者富含参与染色质、转录、RNA生物发生和其他过程的因子。一项更集中的研究表明，DDX47解旋酶是一个R环分解酶，而MeCP2甲基化- cpg结合蛋白揭示了DNA甲基化和R环之间的联系。因此，我们的研究结果表明，过多的基因功能障碍可以通过不同的机制影响r -环稳态来改变细胞生理。一项siRNA筛选鉴定了多达59个影响R环稳态的因子甲基- cpg结合蛋白MeCP2阻止R环和相关的DNA损伤DDX47解绕DNA- rna杂交体并保护细胞免受有害R环的伤害不同细胞过程的功能障碍增加共转录R环Marchena-Cruz等人提出了一项功能测定，确定了59个防止有害R环的因子，这些因子已知参与不同的细胞过程。包括RNA代谢和染色质功能。他们研究了其中的两个因素，DDX47和MeCP2，并得出结论，R环在不相关过程的功能障碍中积累。
Unscheduled R loops can be a source of genome instability, a hallmark of cancer cells. Although targeted proteomic approaches and cellular analysis of specific mutants have uncovered factors potentially involved in R-loop homeostasis, we report a more open screening of factors whose depletion causes R loops based on the ability of activation-induced cytidine deaminase (AID) to target R loops. Immunofluorescence analysis of γH2AX caused by small interfering RNAs (siRNAs) covering 3,205 protein-coding genes identifies 59 potential candidates, from which 13 are analyzed further and show a significant increase of R loops. Such candidates are enriched in factors involved in chromatin, transcription, and RNA biogenesis and other processes. A more focused study shows that the DDX47 helicase is an R-loop resolvase, whereas the MeCP2 methyl-CpG-binding protein uncovers a link between DNA methylation and R loops. Thus, our results suggest that a plethora of gene dysfunctions can alter cell physiology via affecting R-loop homeostasis by different mechanisms. An siRNA screening identifies up to 59 factors affecting R-loop homeostasis The methyl-CpG-binding protein MeCP2 prevents R loops and associated DNA damage DDX47 unwinds DNA-RNA hybrids and protects cells from harmful R loops Dysfunction of different cellular processes increases co-transcriptional R loops Marchena-Cruz et al. present a functional assay identifying 59 factors that protect against harmful R loops and are known to be involved in different cellular processes, including RNA metabolism and chromatin function. They study two of these factors, DDX47 and MeCP2, and conclude that R loops accumulate upon dysfunction of unrelated processes.
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