Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11790, USA.
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11790, USA
Open Biol. 2018 Sep 26;8(9):180110. doi: 10.1098/rsob.180110.
The RNA-binding protein Mei2 is crucial for meiosis in In mutants, pre-meiotic S-phase is blocked, along with meiosis. Mei2 binds a long non-coding RNA (lncRNA) called meiRNA, which is a 'sponge RNA' for the meiotic inhibitor protein Mmi1. The interaction between Mei2, meiRNA and Mmi1 protein is essential for meiosis. But mutants have stronger and different phenotypes than meiRNA mutants, since arrests before pre-meiotic S, while the meiRNA mutant arrests after pre-meiotic S but before meiosis. This suggests Mei2 may bind additional RNAs. To identify novel RNA targets of Mei2, which might explain how Mei2 regulates pre-meiotic S, we used RNA immunoprecipitation and cross-linking immunoprecipitation. In addition to meiRNA, we found the mRNAs for (which encodes Mmi1) and for the S-phase transcription factor There were also three other RNAs of uncertain relevance. We suggest that at meiotic initiation, Mei2 may sequester mRNA to help allow pre-meiotic S, and then may bind both meiRNA and mRNA to inactivate Mmi1 at two levels, the protein level (as previously known), and also the mRNA level, allowing meiosis. We call Mei2-meiRNA a 'double sponge' (i.e. binding both an mRNA and its encoded protein).
RNA 结合蛋白 Mei2 对于减数分裂至关重要。在 突变体中,减数分裂前的 S 期被阻断,同时也阻断了减数分裂。Mei2 结合一种称为 meiRNA 的长非编码 RNA(lncRNA),它是减数分裂抑制剂蛋白 Mmi1 的“海绵 RNA”。Mei2、meiRNA 和 Mmi1 蛋白之间的相互作用对于减数分裂是必不可少的。但 突变体的表型比 meiRNA 突变体更强且不同,因为在减数分裂前的 S 期之前停滞,而 meiRNA 突变体在减数分裂前的 S 期之后但在减数分裂之前停滞。这表明 Mei2 可能结合其他 RNA。为了鉴定 Mei2 的新 RNA 靶标,这些靶标可能解释了 Mei2 如何调节减数分裂前的 S 期,我们使用了 RNA 免疫沉淀和交联免疫沉淀。除了 meiRNA,我们还发现了编码 Mmi1 的 ()mRNA 和 S 期转录因子 的 mRNA。还有三个其他 RNA 的相关性不确定。我们建议,在减数分裂起始时,Mei2 可能将 mRNA 隔离,以帮助启动减数分裂前的 S 期,然后可能结合 meiRNA 和 mRNA,从蛋白水平(如先前所知)和 mRNA 水平,使 Mmi1 失活两个水平,从而允许减数分裂。我们将 Mei2-meiRNA 称为“双重海绵”(即结合一个 mRNA 和其编码的蛋白质)。