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5'非翻译区G-四链体结构以大小依赖的方式增强翻译。

5'UTR G-quadruplex structure enhances translation in size dependent manner.

作者信息

Myong Sua, Lee Chun-Ying, Joshi Meera, Wang Ashley

机构信息

Boston Children's Hospital/Harvard Medical School.

Johns Hopkins University.

出版信息

Res Sq. 2023 Sep 22:rs.3.rs-3352233. doi: 10.21203/rs.3.rs-3352233/v1.

DOI:10.21203/rs.3.rs-3352233/v1
PMID:37790436
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10543253/
Abstract

Translation initiation in bacteria is frequently regulated by various structures in the 5' untranslated region (5'UTR). Previously, we demonstrated that G-quadruplex (G4) formation in non-template DNA enhances transcription. In this study, we aimed to explore how G4 formation in mRNA (RG4) at 5'UTR impacts translation using a T7-based in vitro translation system and in We showed that RG4 strongly promotes translation efficiency in a size-dependent manner. Additionally, inserting a hairpin upstream of the RG4 further enhances translation efficiency, reaching up to a 12-fold increase. We found that the RG4-dependent effect is not due to increased ribosome affinity, ribosome binding site accessibility, or mRNA stability. We proposed a physical barrier model in which bulky structures in 5'UTR prevent ribosome dislodging and thereby increase the translation output. This study provides biophysical insights into the regulatory role of 5'UTR structures in bacterial translation, highlighting their potential applications in tuning gene expression.

摘要

细菌中的翻译起始常常受到5'非翻译区（5'UTR）中各种结构的调控。此前，我们证明非模板DNA中G-四链体（G4）的形成会增强转录。在本研究中，我们旨在利用基于T7的体外翻译系统，探讨5'UTR处mRNA中的G4（RG4）形成如何影响翻译。我们发现RG4以大小依赖的方式强烈促进翻译效率。此外，在RG4上游插入一个发夹结构可进一步提高翻译效率，增幅高达12倍。我们发现RG4依赖性效应并非由于核糖体亲和力增加、核糖体结合位点可及性提高或mRNA稳定性增强。我们提出了一个物理屏障模型，其中5'UTR中的庞大结构可防止核糖体移位，从而增加翻译产出。本研究为5'UTR结构在细菌翻译中的调控作用提供了生物物理见解，突出了它们在调节基因表达方面的潜在应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/64246e49660e/nihpp-rs3352233v1-f0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/8093c96652ae/nihpp-rs3352233v1-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/cd5feeb1787f/nihpp-rs3352233v1-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/ae2e46111392/nihpp-rs3352233v1-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/74a69806559b/nihpp-rs3352233v1-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/7719b17da38e/nihpp-rs3352233v1-f0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/9a1a9998b054/nihpp-rs3352233v1-f0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/64246e49660e/nihpp-rs3352233v1-f0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/8093c96652ae/nihpp-rs3352233v1-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/cd5feeb1787f/nihpp-rs3352233v1-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/ae2e46111392/nihpp-rs3352233v1-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/74a69806559b/nihpp-rs3352233v1-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/7719b17da38e/nihpp-rs3352233v1-f0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/9a1a9998b054/nihpp-rs3352233v1-f0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f801/10543253/64246e49660e/nihpp-rs3352233v1-f0007.jpg

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