Department of Cell and Developmental Biology, John Innes Centre, Norwich Research Park, Norwich, NR4 7UH, UK.
College of Life Sciences, University of Chinese Academy of Sciences, Beijing, 100049, China.
Genome Biol. 2020 Sep 1;21(1):226. doi: 10.1186/s13059-020-02142-9.
Guanine-rich sequences are able to form complex RNA structures termed RNA G-quadruplexes in vitro. Because of their high stability, RNA G-quadruplexes are proposed to exist in vivo and are suggested to be associated with important biological relevance. However, there is a lack of direct evidence for RNA G-quadruplex formation in living eukaryotic cells. Therefore, it is unclear whether any purported functions are associated with the specific sequence content or the formation of an RNA G-quadruplex structure.
Using rG4-seq, we profile the landscape of those guanine-rich regions with the in vitro folding potential in the Arabidopsis transcriptome. We find a global enrichment of RNA G-quadruplexes with two G-quartets whereby the folding potential is strongly influenced by RNA secondary structures. Using in vitro and in vivo RNA chemical structure profiling, we determine that hundreds of RNA G-quadruplex structures are strongly folded in both Arabidopsis and rice, providing direct evidence of RNA G-quadruplex formation in living eukaryotic cells. Subsequent genetic and biochemical analyses show that RNA G-quadruplex folding is able to regulate translation and modulate plant growth.
Our study reveals the existence of RNA G-quadruplex in vivo and indicates that RNA G-quadruplex structures act as important regulators of plant development and growth.
富含鸟嘌呤的序列能够在体外形成复杂的 RNA 结构,称为 RNA G-四链体。由于其高度稳定性,RNA G-四链体被认为存在于体内,并与重要的生物学相关性有关。然而,在活的真核细胞中直接证明 RNA G-四链体形成的证据还很缺乏。因此,尚不清楚任何所谓的功能是否与特定的序列含量或 RNA G-四链体结构的形成有关。
使用 rG4-seq,我们在拟南芥转录组中描绘了具有体外折叠潜力的富含鸟嘌呤区域的图谱。我们发现富含具有两个 G-四联体的 RNA G-四链体的整体富集,其中折叠潜力受 RNA 二级结构的强烈影响。通过体外和体内 RNA 化学结构分析,我们确定在拟南芥和水稻中,数百个 RNA G-四链体结构都强烈折叠,为活的真核细胞中 RNA G-四链体的形成提供了直接证据。随后的遗传和生化分析表明,RNA G-四链体折叠能够调节翻译并调节植物生长。
我们的研究揭示了体内 RNA G-四链体的存在,并表明 RNA G-四链体结构是植物发育和生长的重要调节剂。