Perez Christelle Alexa Garcia, Adachi Shungo, Nong Quang Dang, Adhitama Nikko, Matsuura Tomoaki, Natsume Toru, Wada Tadashi, Kato Yasuhiko, Watanabe Hajime
Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Japan.
Cellular and Molecular Biotechnology Research Institute (CMB), National Institute of Advanced Industrial Science and Technology (AIST), Tokyo, Japan.
PLoS Genet. 2021 Jul 28;17(7):e1009683. doi: 10.1371/journal.pgen.1009683. eCollection 2021 Jul.
Long noncoding RNAs (lncRNAs) are vastly transcribed and extensively studied but lncRNAs overlapping with the sense orientation of mRNA have been poorly studied. We analyzed the lncRNA DAPALR overlapping with the 5´ UTR of the Doublesex1 (Dsx1), the male determining gene in Daphnia magna. By affinity purification, we identified an RNA binding protein, Shep as a DAPALR binding protein. Shep also binds to Dsx1 5´ UTR by recognizing the overlapping sequence and suppresses translation of the mRNA. In vitro and in vivo analyses indicated that DAPALR increased Dsx1 translation efficiency by sequestration of Shep. This regulation was impaired when the Shep binding site in DAPALR was deleted. These results suggest that Shep suppresses the unintentional translation of Dsx1 by setting a threshold; and when the sense lncRNA DAPALR is expressed, DAPALR cancels the suppression caused by Shep. This mechanism may be important to show dimorphic gene expressions such as sex determination and it may account for the binary expression in various developmental processes.
长链非编码RNA（lncRNAs）转录广泛且研究深入，但与mRNA正义链方向重叠的lncRNAs研究较少。我们分析了与大型溞雄性决定基因双性基因1（Dsx1）的5´非翻译区（UTR）重叠的lncRNA DAPALR。通过亲和纯化，我们鉴定出一种RNA结合蛋白Shep作为DAPALR结合蛋白。Shep还通过识别重叠序列与Dsx1 5´UTR结合，并抑制mRNA的翻译。体外和体内分析表明，DAPALR通过隔离Shep提高了Dsx1的翻译效率。当DAPALR中的Shep结合位点被删除时，这种调控就会受损。这些结果表明，Shep通过设定一个阈值来抑制Dsx1的非特异性翻译；当正义lncRNA DAPALR表达时，DAPALR会消除Shep引起的抑制作用。这种机制对于显示性别决定等二态性基因表达可能很重要，并且可能解释了各种发育过程中的二元表达。
