Marhabaie Mohammad, Wharton Tammy H, Kim Sung Yun, Wharton Robin P
bioRxiv. 2023 Oct 24:2023.08.28.555109. doi: 10.1101/2023.08.28.555109.
The translational repressor Nanos (Nos) regulates a single target, maternal hunchback (hb) mRNA, to govern abdominal segmentation in the early Drosophila embryo. Nos is recruited specifically to sites in the 3'-UTR of hb mRNA in collaboration with the sequence-specific RNA-binding protein Pumilio (Pum); on its own, Nos has no binding specificity. Nos is expressed at other stages of development, but very few mRNA targets that might mediate its action at these stages have been described. Nor has it been clear whether Nos is targeted to other mRNAs in concert with Pum or via other mechanisms. In this report, we identify mRNAs targeted by Nos via two approaches. In the first method, we identify mRNAs depleted upon expression of a chimera bearing Nos fused to the nonsense mediated decay (NMD) factor Upf1. We find that, in addition to hb, Upf1-Nos depletes ~2600 mRNAs from the maternal transcriptome in early embryos. Virtually all of these appear to be targeted in a canonical, hb-like manner in concert with Pum. In a second, more conventional approach, we identify mRNAs that are stabilized during the maternal zygotic transition (MZT) in embryos from nos- females. Most (86%) of the 1185 mRNAs regulated by Nos are also targeted by Upf1-Nos, validating use of the chimera. Approximately 60% of mRNAs targeted by Upf1-Nos are not stabilized in the absence of Nos. However, Upf1-Nos mRNA targets are hypo-adenylated and inefficiently translated at the ovary-embryo transition, whether or not they suffer Nos-dependent degradation in the embryo. We suggest that the late ovarian burst of Nos represses a large fraction of the maternal transcriptome, priming it for later degradation by other factors during the MZT in the embryo.
翻译抑制因子Nanos(Nos)调控单个靶标——母体驼背(hb)mRNA,以控制果蝇早期胚胎的腹部体节形成。Nos与序列特异性RNA结合蛋白Pumilio(Pum)协同作用,特异性地募集到hb mRNA 3'-UTR中的位点;单独存在时,Nos没有结合特异性。Nos在发育的其他阶段也有表达,但在这些阶段可能介导其作用的mRNA靶标很少被描述。目前也不清楚Nos是与Pum协同作用靶向其他mRNA,还是通过其他机制靶向。在本报告中,我们通过两种方法鉴定了被Nos靶向的mRNA。在第一种方法中,我们鉴定了在表达与无义介导的衰变(NMD)因子Upf1融合的Nos嵌合体时被耗尽的mRNA。我们发现,除了hb之外,Upf1-Nos在早期胚胎的母体转录组中耗尽了约2600种mRNA。实际上,所有这些mRNA似乎都以与Pum协同的典型hb样方式被靶向。在第二种更传统的方法中,我们鉴定了在nos-雌性胚胎的母源合子转变(MZT)过程中稳定的mRNA。由Nos调控的1185种mRNA中的大多数(86%)也被Upf1-Nos靶向,验证了该嵌合体的用途。在没有Nos的情况下,约60%被Upf1-Nos靶向的mRNA不稳定。然而,无论Upf1-Nos的mRNA靶标在胚胎中是否经历Nos依赖性降解,它们在卵巢-胚胎转变时都会发生低腺苷酸化且翻译效率低下。我们认为,Nos在卵巢后期的爆发抑制了大部分母体转录组,使其在胚胎的MZT过程中为后续被其他因子降解做好准备。