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RelA binding of mRNAs modulates translation or sRNA-mRNA basepairing depending on the position of the GGAG site
Molecular Microbiology ( IF 3.6 ) Pub Date : 2021-09-14 , DOI: 10.1111/mmi.14812
Pallabi Basu 1 , Shoshy Altuvia 1
Affiliation  

Previously, we reported that RelA protein facilitates Hfq-mediated mRNA-sRNA regulation by binding sRNAs carrying a Shine-Dalgarno-like GGAG sequence. In turn, sRNA-Hfq monomers are stabilized, enabling the attachment of more Hfq subunits to form a functional hexamer. Here, using CLIP-seq, we present a global analysis of RelA-bound RNAs showing that RelA interacts with sRNAs as well as with mRNAs carrying a GGAG motif. RelA binding of mRNAs carrying GGAG at position −7 relative to the initiation codon (AUG) inhibits translation by interfering with the binding of 30S ribosomes. The extent of inhibition depends on the distance of GGAG relative to the AUG, as shortening the spacing between GGAG and AUG abrogates RelA-mediated inhibition. Interestingly, RelA binding of target mRNAs carrying GGAG in the coding sequence or close to AUG facilitates target gene regulation by sRNA partners that lack GGAG. However, translation inhibition caused by RelA binding of mRNAs carrying GGAG at position −7 relative to the AUG renders sRNA-mRNA basepairing regulation ineffective. Our study indicates that by binding RNAs carrying GGAG the ribosome-associated RelA protein inhibits translation of specific newly synthesized incoming mRNAs or enables basepairing regulation by their respective sRNA partners, thereby introducing a new regulatory concept for the bacterial response.

中文翻译:

mRNA 的 RelA 结合根据 GGAG 位点的位置调节翻译或 sRNA-mRNA 碱基配对

以前,我们报道了 RelA 蛋白通过结合携带 Shine-Dalgarno 样 GGAG 序列的 sRNA 来促进 Hfq 介导的 mRNA-sRNA 调节。反过来,sRNA-Hfq 单体被稳定,使更多 Hfq 亚基的附着形成功能性六聚体。在这里,我们使用 CLIP-seq 对 RelA 结合的 RNA 进行了全局分析,显示 RelA 与 sRNA 以及携带 GGAG 基序的 mRNA 相互作用。在相对于起始密码子 (AUG) 的位置 -7 携带 GGAG 的 mRNA 的 RelA 结合通过干扰 30S 核糖体的结合来抑制翻译。抑制的程度取决于 GGAG 相对于 AUG 的距离,因为缩短 GGAG 和 AUG 之间的间距会消除 RelA 介导的抑制。有趣的是,在编码序列中携带 GGAG 或接近 AUG 的靶 mRNA 的 RelA 结合促进了缺乏 GGAG 的 sRNA 伴侣对靶基因的调节。然而,在相对于 AUG 的位置 -7 处携带 GGAG 的 mRNA 与 RelA 结合引起的翻译抑制导致 sRNA-mRNA 碱基配对调节无效。我们的研究表明,通过结合携带 GGAG 的 RNA,核糖体相关的 RelA 蛋白会抑制特定新合成的传入 mRNA 的翻译,或者使它们各自的 sRNA 伴侣能够进行碱基配对调节,从而为细菌反应引入新的调节概念。
更新日期:2021-09-14
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