Structural Probing with MNase Tethered to Ribosome Assembly Factors Resolves Flexible RNA Regions within the Nascent Pre-Ribosomal RNA
The synthesis of ribosomes involves the correct folding of the pre-ribosomal RNA within pre-ribosomal particles. The first ribosomal precursor or small subunit processome assembles stepwise on the nascent transcript of the 35S gene. At the earlier stages, the pre-ribosomal particles undergo structur...
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MDPI AG
2022-01-01
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| Series: | Non-Coding RNA |
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| Online Access: | https://www.mdpi.com/2311-553X/8/1/1 |
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| author | Tom Dielforder Christina Maria Braun Fabian Hölzgen Shuang Li Mona Thiele Marina Huber Uli Ohmayer Jorge Perez-Fernandez |
| author_facet | Tom Dielforder Christina Maria Braun Fabian Hölzgen Shuang Li Mona Thiele Marina Huber Uli Ohmayer Jorge Perez-Fernandez |
| author_sort | Tom Dielforder |
| collection | DOAJ |
| description | The synthesis of ribosomes involves the correct folding of the pre-ribosomal RNA within pre-ribosomal particles. The first ribosomal precursor or small subunit processome assembles stepwise on the nascent transcript of the 35S gene. At the earlier stages, the pre-ribosomal particles undergo structural and compositional changes, resulting in heterogeneous populations of particles with highly flexible regions. Structural probing methods are suitable for resolving these structures and providing evidence about the architecture of ribonucleoprotein complexes. Our approach used MNase tethered to the assembly factors Nan1/Utp17, Utp10, Utp12, and Utp13, which among other factors, initiate the formation of the small subunit processome. Our results provide dynamic information about the folding of the pre-ribosomes by elucidating the relative organization of the 5′ETS and ITS1 regions within the 35S and U3 snoRNA around the C-terminal domains of Nan1/Utp17, Utp10, Utp12, and Utp13. |
| first_indexed | 2024-12-19T12:15:00Z |
| format | Article |
| id | doaj.art-75ba1be2daa345b19f37814f74fa6fd2 |
| institution | Directory Open Access Journal |
| issn | 2311-553X |
| language | English |
| last_indexed | 2024-12-19T12:15:00Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
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| series | Non-Coding RNA |
| spelling | doaj.art-75ba1be2daa345b19f37814f74fa6fd22022-12-21T20:22:02ZengMDPI AGNon-Coding RNA2311-553X2022-01-0181110.3390/ncrna8010001Structural Probing with MNase Tethered to Ribosome Assembly Factors Resolves Flexible RNA Regions within the Nascent Pre-Ribosomal RNATom Dielforder0Christina Maria Braun1Fabian Hölzgen2Shuang Li3Mona Thiele4Marina Huber5Uli Ohmayer6Jorge Perez-Fernandez7Department of Biochemistry III, University of Regensburg, Universitätstrasse 31, D-93051 Regensburg, GermanyDepartment of Biochemistry III, University of Regensburg, Universitätstrasse 31, D-93051 Regensburg, GermanyDepartment of Biochemistry III, University of Regensburg, Universitätstrasse 31, D-93051 Regensburg, GermanyDepartment of Biochemistry III, University of Regensburg, Universitätstrasse 31, D-93051 Regensburg, GermanyDepartment of Biochemistry III, University of Regensburg, Universitätstrasse 31, D-93051 Regensburg, GermanyDepartment of Biochemistry III, University of Regensburg, Universitätstrasse 31, D-93051 Regensburg, GermanyDepartment of Biochemistry III, University of Regensburg, Universitätstrasse 31, D-93051 Regensburg, GermanyDepartment of Biochemistry III, University of Regensburg, Universitätstrasse 31, D-93051 Regensburg, GermanyThe synthesis of ribosomes involves the correct folding of the pre-ribosomal RNA within pre-ribosomal particles. The first ribosomal precursor or small subunit processome assembles stepwise on the nascent transcript of the 35S gene. At the earlier stages, the pre-ribosomal particles undergo structural and compositional changes, resulting in heterogeneous populations of particles with highly flexible regions. Structural probing methods are suitable for resolving these structures and providing evidence about the architecture of ribonucleoprotein complexes. Our approach used MNase tethered to the assembly factors Nan1/Utp17, Utp10, Utp12, and Utp13, which among other factors, initiate the formation of the small subunit processome. Our results provide dynamic information about the folding of the pre-ribosomes by elucidating the relative organization of the 5′ETS and ITS1 regions within the 35S and U3 snoRNA around the C-terminal domains of Nan1/Utp17, Utp10, Utp12, and Utp13.https://www.mdpi.com/2311-553X/8/1/1ribosome biogenesisSSU-processome<i>S. cerevisiae</i>structural probingRNA |
| spellingShingle | Tom Dielforder Christina Maria Braun Fabian Hölzgen Shuang Li Mona Thiele Marina Huber Uli Ohmayer Jorge Perez-Fernandez Structural Probing with MNase Tethered to Ribosome Assembly Factors Resolves Flexible RNA Regions within the Nascent Pre-Ribosomal RNA Non-Coding RNA ribosome biogenesis SSU-processome <i>S. cerevisiae</i> structural probing RNA |
| title | Structural Probing with MNase Tethered to Ribosome Assembly Factors Resolves Flexible RNA Regions within the Nascent Pre-Ribosomal RNA |
| title_full | Structural Probing with MNase Tethered to Ribosome Assembly Factors Resolves Flexible RNA Regions within the Nascent Pre-Ribosomal RNA |
| title_fullStr | Structural Probing with MNase Tethered to Ribosome Assembly Factors Resolves Flexible RNA Regions within the Nascent Pre-Ribosomal RNA |
| title_full_unstemmed | Structural Probing with MNase Tethered to Ribosome Assembly Factors Resolves Flexible RNA Regions within the Nascent Pre-Ribosomal RNA |
| title_short | Structural Probing with MNase Tethered to Ribosome Assembly Factors Resolves Flexible RNA Regions within the Nascent Pre-Ribosomal RNA |
| title_sort | structural probing with mnase tethered to ribosome assembly factors resolves flexible rna regions within the nascent pre ribosomal rna |
| topic | ribosome biogenesis SSU-processome <i>S. cerevisiae</i> structural probing RNA |
| url | https://www.mdpi.com/2311-553X/8/1/1 |
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