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<p>The most common rRNA modification is 2’-<i>O</i>-methylation. Here, we determine the landscape of 2’-<i>O</i>-methylation in <i>Leishmania</i>, a parasite that cycles between two different hosts, insect and mammalian. We find two 2’-<i>O</i>-methylated positions that are differentially modified during the parasite’s two life stages. The deposition of these modifications is guided by snoRNAs. When we perform cytosine base-editing of the snoRNA responsible for guiding one of the two stage-regulated modifications, Am479, we fail to detect ribosomes lacking this modification, suggesting that it is essential. To better understand the role of the snoRNA and its guided modification, we determine the cryo-EM structures of ribosomes from cells overexpressing the guiding snoRNA and compare them to ribosomes from the parental strain. We do not find structural changes around Am479 or in the small subunit rRNA, but observe a difference in H68 of the large subunit rRNA due to a second base-pairing interaction, suggesting a potential chaperone activity for the snoRNA. Based on these results, translatome and tRNA analysis, we propose a mechanism whereby changes in ribosome structure affect the release of specific tRNAs, which correlate with changes in translation of only a subset of mRNAs.</p>

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A small nucleolar RNA dictates the structure and function of translating ribosomes in Leishmania

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月13日

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  • K. Shanmugha Rajan,
  • Saurav Aryal,
  • Sharanya Murugeshan,
  • Yinzhou Zhu,
  • Anat Bashan,
  • Mika Olami,
  • Hava Madmoni,
  • Yuko Nobe,
  • Tirza Doniger,
  • Smadar Cohen-Chalamish,
  • Eric Prina,
  • Pascale Pescher,
  • Tom Beneke,
  • Masato Taoka,
  • Christopher L. Holley,
  • Ron Unger,
  • Gerald F. Späth,
  • Ada Yonath,
  • Shulamit Michaeli

摘要

The most common rRNA modification is 2’-O-methylation. Here, we determine the landscape of 2’-O-methylation in Leishmania, a parasite that cycles between two different hosts, insect and mammalian. We find two 2’-O-methylated positions that are differentially modified during the parasite’s two life stages. The deposition of these modifications is guided by snoRNAs. When we perform cytosine base-editing of the snoRNA responsible for guiding one of the two stage-regulated modifications, Am479, we fail to detect ribosomes lacking this modification, suggesting that it is essential. To better understand the role of the snoRNA and its guided modification, we determine the cryo-EM structures of ribosomes from cells overexpressing the guiding snoRNA and compare them to ribosomes from the parental strain. We do not find structural changes around Am479 or in the small subunit rRNA, but observe a difference in H68 of the large subunit rRNA due to a second base-pairing interaction, suggesting a potential chaperone activity for the snoRNA. Based on these results, translatome and tRNA analysis, we propose a mechanism whereby changes in ribosome structure affect the release of specific tRNAs, which correlate with changes in translation of only a subset of mRNAs.