Department of Life Science, Graduate School of Advanced Engineering, Chiba Institute of Technology, Chiba 275-0016, Japan.
Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo 113-0032, Japan.
RNA. 2022 Apr;28(4):541-550. doi: 10.1261/rna.078967.121. Epub 2022 Jan 5.
PIWI-interacting RNAs (piRNAs) repress transposons to protect the germline genome from DNA damage caused by transposon transposition. In , the () mRNA is consumed to produce piRNA in its 3'-UTR. A element located within the 3'-UTR, , is necessary for piRNA biogenesis. In this study, we analyzed the structure of the RNA, a 100-nt RNA corresponding to the element, by the SHAPE and NMR analyses and found that a stable hairpin structure formed in the 5' half of the RNA. The tertiary structure of the 16-nt stable hairpin was analyzed by NMR, and a novel stem-loop structure, the T-hairpin, was found. In the T-hairpin, four uridine residues are exposed to the solvent, suggesting that this stem-loop is the target of Yb protein, a Tudor domain-containing piRNA biogenesis factor. The piRNA biogenesis assay showed that both the T-hairpin and the 3' half are required for the function of the element, suggesting that both the T-hairpin and the 3' half are recognized by Yb protein.
PIWI 相互作用 RNA（piRNA）抑制转座子，以保护生殖系基因组免受转座子转座引起的 DNA 损伤。在 中，mRNA 被消耗以在其 3'-UTR 中产生 piRNA。位于 3'-UTR 内的 元件对于 piRNA 的生物发生是必需的。在这项研究中，我们通过 SHAPE 和 NMR 分析分析了 RNA 的结构，该 RNA 是对应于 元件的 100nt RNA，并发现 RNA 的 5' 半部分形成了一个稳定的发夹结构。通过 NMR 分析了 16nt 稳定发夹的三级结构，发现了一个新的茎环结构，即 T 发夹。在 T 发夹中，四个尿嘧啶残基暴露在溶剂中，这表明该茎环是 Tudor 结构域包含的 piRNA 生物发生因子 Yb 蛋白的靶标。piRNA 生物发生测定表明，T 发夹和 3' 半部分对于 元件的功能都是必需的，这表明 T 发夹和 3' 半部分都被 Yb 蛋白识别。
