Scholarlink

<p>Super-resolution imaging with RhoBAST fluorogenic RNA aptamer sheds new light on decoding RNA dynamics with high spatiotemporal resolution. To uncover the molecular basis underlying its exceptional photophysical properties, we determined the first free-form structure of RhoBAST, along with its ligand-bound complexes with TMR-DN and related analogs. RhoBAST adopts an inverted ‘V’-shaped architecture, with the fluorophore accommodated between the J2/3 and L3 loops. Comparative structural analyses reveal that nucleotide G38 undergoes a pronounced conformational transition—from an inward-facing to an outward-flipped orientation—upon ligand binding. Structure-guided mutagenesis and complementary biophysical assays, including fluorescence spectroscopy, surface plasmon resonance, and 2-aminopurine kinetics, establish that this dynamic nucleotide flipping facilitates rapid ligand turnover and fluorescence flickering, the hallmark of super-resolution imaging. These findings uncover a previously unrecognized nucleotide-flipping mechanism in a fluorogenic RNA aptamer, providing a structural and mechanistic blueprint for the rational design of next-generation super-resolution RNA imaging tools.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Nucleotide flipping correlates with fluorescence activation in the RhoBAST imaging platform

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年7月21日

查看全文
  • Xiaoqing Tai,
  • Mengqi He,
  • Christoph Mitteregger,
  • Huiqin You,
  • Zejia Hu,
  • Peng Xiong,
  • Xinyue Bao,
  • Xin Shen,
  • Hongcheng Li,
  • Yu Li,
  • Jinzhu Zhang,
  • Ronald Micura,
  • Aiming Ren

摘要

Super-resolution imaging with RhoBAST fluorogenic RNA aptamer sheds new light on decoding RNA dynamics with high spatiotemporal resolution. To uncover the molecular basis underlying its exceptional photophysical properties, we determined the first free-form structure of RhoBAST, along with its ligand-bound complexes with TMR-DN and related analogs. RhoBAST adopts an inverted ‘V’-shaped architecture, with the fluorophore accommodated between the J2/3 and L3 loops. Comparative structural analyses reveal that nucleotide G38 undergoes a pronounced conformational transition—from an inward-facing to an outward-flipped orientation—upon ligand binding. Structure-guided mutagenesis and complementary biophysical assays, including fluorescence spectroscopy, surface plasmon resonance, and 2-aminopurine kinetics, establish that this dynamic nucleotide flipping facilitates rapid ligand turnover and fluorescence flickering, the hallmark of super-resolution imaging. These findings uncover a previously unrecognized nucleotide-flipping mechanism in a fluorogenic RNA aptamer, providing a structural and mechanistic blueprint for the rational design of next-generation super-resolution RNA imaging tools.