Structural and mechanistic insights into ribosomal ITS2 RNA processing by nuclease-kinase machinery
Precursor ribosomal RNA (pre-rRNA) processing is a key step in ribosome biosynthesis and involves numerous RNases. A HEPN (higher eukaryote and prokaryote nucleotide binding) nuclease Las1 and a polynucleotide kinase Grc3 assemble into a tetramerase responsible for rRNA maturation. Here, we report t...
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eLife Sciences Publications Ltd
2024-01-01
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| 丛编: | eLife |
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| 在线阅读: | https://elifesciences.org/articles/86847 |
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| author | Jiyun Chen Hong Chen Shanshan Li Xiaofeng Lin Rong Hu Kaiming Zhang Liang Liu |
| author_facet | Jiyun Chen Hong Chen Shanshan Li Xiaofeng Lin Rong Hu Kaiming Zhang Liang Liu |
| author_sort | Jiyun Chen |
| collection | DOAJ |
| description | Precursor ribosomal RNA (pre-rRNA) processing is a key step in ribosome biosynthesis and involves numerous RNases. A HEPN (higher eukaryote and prokaryote nucleotide binding) nuclease Las1 and a polynucleotide kinase Grc3 assemble into a tetramerase responsible for rRNA maturation. Here, we report the structures of full-length Saccharomyces cerevisiae and Cyberlindnera jadinii Las1-Grc3 complexes, and C. jadinii Las1. The Las1-Grc3 structures show that the central coiled-coil domain of Las1 facilitates pre-rRNA binding and cleavage, while the Grc3 C-terminal loop motif directly binds to the HEPN active center of Las1 and regulates pre-rRNA cleavage. Structural comparison between Las1 and Las1-Grc3 complex exhibits that Grc3 binding induces conformational rearrangements of catalytic residues associated with HEPN nuclease activation. Biochemical assays identify that Las1 processes pre-rRNA at the two specific sites (C2 and C2′), which greatly facilitates rRNA maturation. Our structures and specific pre-rRNA cleavage findings provide crucial insights into the mechanism and pathway of pre-rRNA processing in ribosome biosynthesis. |
| first_indexed | 2024-03-08T16:37:36Z |
| format | Article |
| id | doaj.art-d13151944a24497ca72e50ec1a8350c2 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-03-08T16:37:36Z |
| publishDate | 2024-01-01 |
| publisher | eLife Sciences Publications Ltd |
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| series | eLife |
| spelling | doaj.art-d13151944a24497ca72e50ec1a8350c22024-01-05T14:10:57ZengeLife Sciences Publications LtdeLife2050-084X2024-01-011210.7554/eLife.86847Structural and mechanistic insights into ribosomal ITS2 RNA processing by nuclease-kinase machineryJiyun Chen0Hong Chen1https://orcid.org/0000-0002-0179-4493Shanshan Li2Xiaofeng Lin3Rong Hu4Kaiming Zhang5Liang Liu6https://orcid.org/0000-0002-5379-0638State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University, Xiamen, ChinaState Key Laboratory of Cellular Stress Biology, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University, Xiamen, ChinaMOE Key Laboratory for Cellular Dynamics and Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, ChinaState Key Laboratory of Cellular Stress Biology, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University, Xiamen, ChinaState Key Laboratory of Cellular Stress Biology, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University, Xiamen, ChinaMOE Key Laboratory for Cellular Dynamics and Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, ChinaState Key Laboratory of Cellular Stress Biology, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University, Xiamen, ChinaPrecursor ribosomal RNA (pre-rRNA) processing is a key step in ribosome biosynthesis and involves numerous RNases. A HEPN (higher eukaryote and prokaryote nucleotide binding) nuclease Las1 and a polynucleotide kinase Grc3 assemble into a tetramerase responsible for rRNA maturation. Here, we report the structures of full-length Saccharomyces cerevisiae and Cyberlindnera jadinii Las1-Grc3 complexes, and C. jadinii Las1. The Las1-Grc3 structures show that the central coiled-coil domain of Las1 facilitates pre-rRNA binding and cleavage, while the Grc3 C-terminal loop motif directly binds to the HEPN active center of Las1 and regulates pre-rRNA cleavage. Structural comparison between Las1 and Las1-Grc3 complex exhibits that Grc3 binding induces conformational rearrangements of catalytic residues associated with HEPN nuclease activation. Biochemical assays identify that Las1 processes pre-rRNA at the two specific sites (C2 and C2′), which greatly facilitates rRNA maturation. Our structures and specific pre-rRNA cleavage findings provide crucial insights into the mechanism and pathway of pre-rRNA processing in ribosome biosynthesis.https://elifesciences.org/articles/86847HEPN nucleasePre-rRNA processingribosome biosynthesis |
| spellingShingle | Jiyun Chen Hong Chen Shanshan Li Xiaofeng Lin Rong Hu Kaiming Zhang Liang Liu Structural and mechanistic insights into ribosomal ITS2 RNA processing by nuclease-kinase machinery eLife HEPN nuclease Pre-rRNA processing ribosome biosynthesis |
| title | Structural and mechanistic insights into ribosomal ITS2 RNA processing by nuclease-kinase machinery |
| title_full | Structural and mechanistic insights into ribosomal ITS2 RNA processing by nuclease-kinase machinery |
| title_fullStr | Structural and mechanistic insights into ribosomal ITS2 RNA processing by nuclease-kinase machinery |
| title_full_unstemmed | Structural and mechanistic insights into ribosomal ITS2 RNA processing by nuclease-kinase machinery |
| title_short | Structural and mechanistic insights into ribosomal ITS2 RNA processing by nuclease-kinase machinery |
| title_sort | structural and mechanistic insights into ribosomal its2 rna processing by nuclease kinase machinery |
| topic | HEPN nuclease Pre-rRNA processing ribosome biosynthesis |
| url | https://elifesciences.org/articles/86847 |
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