Fam208a orchestrates interaction protein network essential for early embryonic development and cell division (Retracted Article)
Fam208a orchestrates interaction protein network essential for early embryonic development and cell division (Retracted Article)
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DOI：
10.1016/j.yexcr.2019.05.018
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发表时间：
2019-09-01
影响因子：
3.7
通讯作者：
Sedlacek, Radislav
中科院分区：
文献类型：
--
作者：
Gresakova, Veronika;Novosadova, Vendula;Sedlacek, Radislav
基因组稳定性的维持对于每个活细胞都是必不可少的，因为遗传信息在每次细胞分裂事件中的DNA复制期间反复受到挑战。有丝分裂或减数分裂期间出现的错误、缺陷、延迟和错误会导致DNA修复过程的激活，如果修复过程失败，可能会启动程序性细胞死亡，即细胞凋亡。Fam 208 a是一种蛋白质，其在异染色质维持中的重要性最近已被描述。在这项工作中，我们描述了Fam 208 a在细胞分裂过程中维持基因组稳定性的关键作用。使用CRISPR/Cas9在小鼠中靶向消耗Fam 208 a导致E12.5之前的胚胎死亡。我们还使用siRNA方法下调受精卵中的Fam 208 a，以避免发育早期阶段母体RNA的影响。这种早期下调增加了在两细胞阶段胚胎发育的停滞和多极纺锤体形成的发生。为了进一步研究这一点，我们使用酵母双杂交（Y2 H）系统，并确定了新的推定的相互作用伙伴Gpsm 2，Amn 1，Eml 1，Svil，和Itgb 3bp。在多种鼠组织中通过qRT-PCR分析和原位杂交[1]评估它们与Fam 208 a的共表达。基于这些结果，我们提出Fam 208 a通过与Mphosph 8相互作用在HUSH复合物中发挥作用，因为这些蛋白质不仅能够物理相互作用，而且还能够共定位。我们带来了新的证据，证明Fam 208 a是一种多相互作用的蛋白质，在发育的最早阶段影响细胞分裂水平上的基因组稳定性，也通过与成年组织中的甲基化复合物相互作用影响基因组稳定性。除了其表观遗传功能，Fam 208 a似乎在合子分裂中具有额外的作用，可能通过与新鉴定的推定伴侣Gpsm 2，Amn 1，Eml 1，Svil和Itgb 3bp相互作用。
Maintenance of genome stability is essential for every living cell as genetic information is repeatedly challenged during DNA replication in each cell division event. Errors, defects, delays, and mistakes that arise during mitosis or meiosis lead to an activation of DNA repair processes and in case of their failure, programmed cell death, i.e. apoptosis, could be initiated. Fam208a is a protein whose importance in heterochromatin maintenance has been described recently. In this work, we describe the crucial role of Fam208a in sustaining the genome stability during the cellular division. The targeted depletion of Fam208a in mice using CRISPR/Cas9 leads to embryonic lethality before E12.5. We also used the siRNA approach to downregulate Fam208a in zygotes to avoid the influence of maternal RNA in the early stages of development. This early downregulation increased arresting of the embryonal development at the two-cell stage and occurrence of multipolar spindles formation. To investigate this further, we used the yeast two-hybrid (Y2H) system and identified new putative interaction partners Gpsm2, Amn1, Eml1, Svil, and Itgb3bp. Their co-expression with Fam208a was assessed by qRT-PCR profiling and in situ hybridisation [1] in multiple murine tissues. Based on these results we proposed that Fam208a functions within the HUSH complex by interaction with Mphosph8 as these proteins are not only able to physically interact but also co-localise. We are bringing new evidence that Fam208a is multi-interacting protein affecting genome stability on the level of cell division at the earliest stages of development and also by interaction with methylation complex in adult tissues. In addition to its epigenetic functions, Fam208a appears to have an additional role in zygotic division, possibly via interaction with newly identified putative partners Gpsm2, Amn1, Eml1, Svil, and Itgb3bp.