Scholarlink

<p>Enigmatic dinucleoside tetraphosphates, known as ‘alarmones’ (Np<sub>4</sub>Ns), have recently been shown to function in bacteria as precursors to Np<sub>4</sub> caps on transcripts, likely influencing RNA longevity and cellular adaptation to stress. In proteobacteria, ApaH is the predominant enzyme that hydrolyzes Np<sub>4</sub>Ns and decaps Np<sub>4</sub>-capped RNAs to initiate their 5′-end-dependent degradation. Here we conducted a biochemical and structural study to uncover the catalytic mechanism of <i>Escherichia coli</i> ApaH, a prototypic symmetric Np<sub>4</sub>N hydrolase, on various Np<sub>4</sub>Ns and Np<sub>4</sub>-capped RNAs. We found that the enzyme uses a unique combination of nonspecific and semispecific substrate recognition, enabling substrates to bind in two orientations with a slight orientational preference. Despite such exceptional recognition properties, ApaH efficiently decaps various Np<sub>4</sub>-capped mRNAs and sRNAs, thereby impacting their lifetimes. Our findings highlight the need to determine substrate orientation preferences before designing substrate-mimicking drugs, as enzymes may escape activity modulation with one of the alternative substrate orientations.</p><p></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

ApaH decaps Np4N-capped RNAs in two alternative orientations

  • 期刊论文

  • 发表日期：

    2025年8月11日

查看全文
  • Ashok Nuthanakanti,
  • Megan Korn,
  • Rose Levenson-Palmer,
  • Yue Wu,
  • Nandhini Rajesh Babu,
  • Xuhui Huang,
  • Robert S. Banh,
  • Joel G. Belasco,
  • Alexander Serganov

摘要

Enigmatic dinucleoside tetraphosphates, known as ‘alarmones’ (Np4Ns), have recently been shown to function in bacteria as precursors to Np4 caps on transcripts, likely influencing RNA longevity and cellular adaptation to stress. In proteobacteria, ApaH is the predominant enzyme that hydrolyzes Np4Ns and decaps Np4-capped RNAs to initiate their 5′-end-dependent degradation. Here we conducted a biochemical and structural study to uncover the catalytic mechanism of Escherichia coli ApaH, a prototypic symmetric Np4N hydrolase, on various Np4Ns and Np4-capped RNAs. We found that the enzyme uses a unique combination of nonspecific and semispecific substrate recognition, enabling substrates to bind in two orientations with a slight orientational preference. Despite such exceptional recognition properties, ApaH efficiently decaps various Np4-capped mRNAs and sRNAs, thereby impacting their lifetimes. Our findings highlight the need to determine substrate orientation preferences before designing substrate-mimicking drugs, as enzymes may escape activity modulation with one of the alternative substrate orientations.