Nakajima Ryusuke, Sato Takuya, Ogawa Takehiko, Okano Hideyuki, Noce Toshiaki
Department of Physiology, Keio University School of Medicine, 35 Shinamomachi, Shinjuku-ku, Tokyo, Japan.
Laboratory of Proteomics, Institute of Molecular Medicine and Life Science, Yokohama City University Association of Medical Science, Yokohama, Kanagawa, Japan.
PLoS One. 2017 Jun 28;12(6):e0179585. doi: 10.1371/journal.pone.0179585. eCollection 2017.
A search for early response genes that are activated following germ cell induction from mouse embryonic stem cells in vitro led us to the isolation of a long noncoding RNA that contains a SINE (short interspersed element)-B1F motif that was named R53. In situ hybridization and northern blot analyses revealed that the R53 subfragment RNA bears a B1F motif, is processed from the primary transcript, is expressed in adult testis and is predominantly localized in meiotic metaphase chromatin during spermatogenesis. Recent studies of chromosome-associated RNAs have explored novel functions of noncoding RNAs. Specifically, chromosome-bound noncoding RNAs function not only as structural components of chromosome but also as scaffolds that recruit epigenetic modulators for transcriptional regulation, and they are dynamically rearranged during the cell cycle. However, few studies have explored meiotic chromatin; thus, R53 RNA appears to be the first long noncoding RNA to be tightly associated with the metaphase chromatin during spermatogenesis. Furthermore, R53 knockdown using a lentivirus-mediated RNAi injected into mouse testis and organ culture of the fragments revealed a remarkable reduction in postmeiotic cells and irregular up-regulation of several postmeiotic genes, which suggests the possibility that the SINE-B1-derived noncoding RNA R53 plays an indispensable role in the transcriptional regulation of key spermatogenesis genes.
为了寻找从小鼠胚胎干细胞体外诱导生殖细胞后被激活的早期反应基因,我们分离出了一种长链非编码RNA,它含有一个SINE(短散在元件)-B1F基序,被命名为R53。原位杂交和Northern印迹分析表明,R53亚片段RNA带有B1F基序,是从初级转录本加工而来,在成年睾丸中表达,并且在精子发生过程中主要定位于减数分裂中期染色质。最近对染色体相关RNA的研究探索了非编码RNA的新功能。具体而言,与染色体结合的非编码RNA不仅作为染色体的结构成分,还作为招募表观遗传调节剂进行转录调控的支架,并且它们在细胞周期中动态重排。然而,很少有研究探索减数分裂染色质;因此,R53 RNA似乎是第一个在精子发生过程中与中期染色质紧密相关的长链非编码RNA。此外,将慢病毒介导的RNAi注射到小鼠睾丸中敲低R53,并对片段进行器官培养,结果显示减数分裂后细胞显著减少,几个减数分裂后基因出现不规则上调,这表明源自SINE-B1的非编码RNA R53可能在关键精子发生基因的转录调控中发挥不可或缺的作用。