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Antisense RNA protects mRNA from RNase E degradation by RNA–RNA duplex formation during phage infection

机译:反义RNA通过在噬菌体感染过程中通过RNA-RNA双链体的形成来保护mRNA免受RNAase E的降解

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摘要

The ecologically important cyanobacterium Prochlorococcus possesses the smallest genome among oxyphototrophs, with a reduced suite of protein regulators and a disproportionately high number of regulatory RNAs. Many of these are asRNAs, raising the question whether they modulate gene expression through the protection of mRNA from RNase E degradation. To address this question, we produced recombinant RNase E from Prochlorococcus sp. MED4, which functions optimally at 12 mM Mg2+, pH 9 and 35°C. RNase E cleavage assays were performed with this recombinant protein to assess enzyme activity in the presence of single- or double-stranded RNA substrates. We found that extraordinarily long asRNAs of 3.5 and 7 kb protect a set of mRNAs from RNase E degradation that accumulate during phage infection. These asRNA–mRNA duplex formations mask single-stranded recognition sites of RNase E, leading to increased stability of the mRNAs. Such interactions directly modulate RNA stability and provide an explanation for enhanced transcript abundance of certain mRNAs during phage infection. Protection from RNase E-triggered RNA decay may constitute a hitherto unknown regulatory function of bacterial cis-asRNAs, impacting gene expression.
机译:具有重要生态意义的蓝藻原球菌在氧养生物中拥有最小的基因组,其蛋白质调节剂减少了,而调节RNA的数量却成比例地增加。其中许多是asRNA,提出了一个问题,即它们是否通过保护mRNA免受RNase E降解来调节基因表达。为了解决这个问题,我们从Prochlorococcus sp。生产了重组RNaseE。 MED4在12 mM Mg 2 + ,pH 9和35°C时表现最佳。用这种重组蛋白进行RNase E裂解测定,以评估在单链或双链RNA底物存在下的酶活性。我们发现异常长的3.5和7kb的asRNA保护了一组在噬菌体感染期间积累的RNase E降解mRNA。这些asRNA-mRNA双链体形成掩盖了RNase E的单链识别位点,从而提高了mRNA的稳定性。此类相互作用直接调节RNA的稳定性,并为噬菌体感染过程中某些mRNA的转录本丰度提高提供了解释。保护免受RNase E触发的RNA衰变可能构成细菌顺式asRNA迄今未知的调节功能,从而影响基因表达。

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