Identification of genes that promote or antagonize somatic homolog pairing using a high-throughput FISH-based screen.
Identification of genes that promote or antagonize somatic homolog pairing using a high-throughput FISH-based screen.
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DOI：
10.1371/journal.pgen.1002667
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发表时间：
2012
期刊：
影响因子：
4.5
通讯作者：
Wu CT
中科院分区：
文献类型：
--
作者：
Joyce EF;Williams BR;Xie T;Wu CT
同源染色体的配对是减数分裂细胞的基本特征。此外，许多物种在非减数分裂体细胞中也表现出同源配对，有证据表明其对基因调控和双链断裂（DSB）修复的影响。体细胞配对的一个极端例子可以在黑腹果蝇中观察到，其中同源染色体在大部分发育过程中保持对齐。然而，我们对体细胞同源配对机制的理解仍然不清楚，因为只有少数基因参与了这一过程。在这项研究中，我们介绍了一种新的高通量荧光原位杂交（FISH）技术，使我们能够进行全基因组RNAi屏幕中观察到的强大的果蝇体细胞配对的因素。我们确定了候选的“配对促进基因”和候选的“反配对基因”，提供了配对是一个动态过程的证据，可以增强和拮抗。许多被发现对促进配对很重要的基因高度富集了与有丝分裂细胞分裂相关的功能，这表明有丝分裂期间染色体动态与间期核组织之间存在长期联系的遗传框架。相反，几个候选的反配对基因具有已知的间期功能与S期进展，DNA复制，染色质压实，包括几个组件的凝聚素II复合物。结合各种次级测定，这些结果提供了深入了解体细胞配对的机制和动力学。除了它们的数量和结构之外，染色体的位置和空间动态也受到严格控制，因为染色体之间的直接相互作用可以促进基因的激活或抑制。在这里，我们专注于一种特殊类型的相互作用，称为体细胞同源配对，发生在染色体的母系和父系副本之间。虽然体细胞配对在下游同源性驱动过程中的作用已经得到了很好的确立，但关于同源染色体片段如何在体细胞内找到彼此、物理对齐并形成稳定的配对相互作用，还有很多东西需要了解。利用一种新的高通量FISH技术和同源染色体密切配对沿着它们的长度在果蝇的体细胞的事实，我们进行了筛选的因素，是重要的体细胞配对的保真度。最终，这些配对基因的特征将揭示配对的机制，以及对发育和疾病有影响的配对介导的过程。最后，我们的配对基因筛选的功效表明，这里描述的高通量FISH技术将被证明是有用的研究形式的核组织和染色体定位配对以外。
The pairing of homologous chromosomes is a fundamental feature of the meiotic cell. In addition, a number of species exhibit homolog pairing in nonmeiotic, somatic cells as well, with evidence for its impact on both gene regulation and double-strand break (DSB) repair. An extreme example of somatic pairing can be observed in Drosophila melanogaster, where homologous chromosomes remain aligned throughout most of development. However, our understanding of the mechanism of somatic homolog pairing remains unclear, as only a few genes have been implicated in this process. In this study, we introduce a novel high-throughput fluorescent in situ hybridization (FISH) technology that enabled us to conduct a genome-wide RNAi screen for factors involved in the robust somatic pairing observed in Drosophila. We identified both candidate “pairing promoting genes” and candidate “anti-pairing genes,” providing evidence that pairing is a dynamic process that can be both enhanced and antagonized. Many of the genes found to be important for promoting pairing are highly enriched for functions associated with mitotic cell division, suggesting a genetic framework for a long-standing link between chromosome dynamics during mitosis and nuclear organization during interphase. In contrast, several of the candidate anti-pairing genes have known interphase functions associated with S-phase progression, DNA replication, and chromatin compaction, including several components of the condensin II complex. In combination with a variety of secondary assays, these results provide insights into the mechanism and dynamics of somatic pairing. In addition to their number and structure, the position and spatial dynamics of chromosomes are under tight control, as direct interactions between chromosomes can contribute to the activation or repression of genes. Here, we focus on a particular type of interaction, known as somatic homolog pairing, which occurs between the maternal and paternal copies of chromosomes. While the role of somatic pairing on downstream homology-driven processes is well-established, there is much to be learned about how homologous chromosome segments find each other, physically align, and form stable pairing interactions within somatic cells. Taking advantage of a novel high-throughput FISH technology and the fact that homologous chromosomes are intimately paired along their lengths in the somatic cells of Drosophila, we have conducted a screen for factors that are important for the fidelity of somatic pairing. Ultimately, the characterization of these pairing genes will shed light on the mechanism of pairing, as well as pairing-mediated processes that have implications for development and disease. Finally, the efficacy of our screen for pairing genes suggests that the high-throughput FISH technology described here will prove useful for studying forms of nuclear organization and chromosome positioning beyond pairing.
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影响因子：
4
作者：
Delcros, Jean-Guy;Prigent, Claude;Giet, Regis
通讯作者：
Giet, Regis
DOI：
10.1083/jcb.200908026
发表时间：
2010-02-22
期刊：
The Journal of cell biology
影响因子：
--
作者：
Fazzio TG;Panning B
通讯作者：
Panning B
影响因子：
4
作者：
Dorsett, Dale
通讯作者：
Dorsett, Dale
影响因子：
64.8
作者：
Björklund, M;Taipale, M;Taipale, J
通讯作者：
Taipale, J
影响因子：
3.3
作者：
Bateman, Jack R.;Wu, C. -ting
通讯作者：
Wu, C. -ting