Structural and functional analysis of a plant nucleolar RNA chaperone-like protein
Abstract Ribosome biogenesis is a key process in all eukaryotic cells that requires hundreds of ribosome biogenesis factors (RBFs), which are essential to build the mature ribosomes consisting of proteins and rRNAs. The processing of the required rRNAs has been studied extensively in yeast and mamma...
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Nature Portfolio
2023-06-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-36426-4 |
| _version_ | 1797801455090925568 |
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| author | Rita Fernandes Anna Ostendorp Steffen Ostendorp Judith Mehrmann Sven Falke Melissa Ann Graewert Magdalena Weingartner Julia Kehr Stefan Hoth |
| author_facet | Rita Fernandes Anna Ostendorp Steffen Ostendorp Judith Mehrmann Sven Falke Melissa Ann Graewert Magdalena Weingartner Julia Kehr Stefan Hoth |
| author_sort | Rita Fernandes |
| collection | DOAJ |
| description | Abstract Ribosome biogenesis is a key process in all eukaryotic cells that requires hundreds of ribosome biogenesis factors (RBFs), which are essential to build the mature ribosomes consisting of proteins and rRNAs. The processing of the required rRNAs has been studied extensively in yeast and mammals, but in plants much is still unknown. In this study, we focused on a RBF from A. thaliana that we named NUCLEOLAR RNA CHAPERONE-LIKE 1 (NURC1). NURC1 was localized in the nucleolus of plant cell nuclei, and other plant RBF candidates shared the same localization. SEC-SAXS experiments revealed that NURC1 has an elongated and flexible structure. In addition, SEC-MALLS experiments confirmed that NURC1 was present in its monomeric form with a molecular weight of around 28 kDa. RNA binding was assessed by performing microscale thermophoresis with the Arabidopsis internal transcribed spacer 2 (ITS2) of the polycistronic pre-rRNA precursor, which contains the 5.8S, 18S, and 25S rRNA. NURC1 showed binding activity to the ITS2 with a dissociation constant of 228 nM and exhibited RNA chaperone-like activity. Our data suggested that NURC1 may have a function in pre-rRNA processing and thus ribosome biogenesis. |
| first_indexed | 2024-03-13T04:50:44Z |
| format | Article |
| id | doaj.art-9f04ef727caa41c6b9c71e3443d81980 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-13T04:50:44Z |
| publishDate | 2023-06-01 |
| publisher | Nature Portfolio |
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| spelling | doaj.art-9f04ef727caa41c6b9c71e3443d819802023-06-18T11:14:04ZengNature PortfolioScientific Reports2045-23222023-06-0113111210.1038/s41598-023-36426-4Structural and functional analysis of a plant nucleolar RNA chaperone-like proteinRita Fernandes0Anna Ostendorp1Steffen Ostendorp2Judith Mehrmann3Sven Falke4Melissa Ann Graewert5Magdalena Weingartner6Julia Kehr7Stefan Hoth8Molecular Plant Physiology, Institute of Plant Science and Microbiology, Department of Biology, Universität HamburgMolecular Plant Genetics, Institute of Plant Science and Microbiology, Department of Biology, Universität HamburgMolecular Plant Genetics, Institute of Plant Science and Microbiology, Department of Biology, Universität HamburgMolecular Plant Physiology, Institute of Plant Science and Microbiology, Department of Biology, Universität HamburgCenter for Free-Electron Laser Science (CFEL), Deutsches Elektronen Synchrotron (DESY)European Molecular Biology Laboratory (EMBL) Hamburg UnitMolecular Plant Physiology, Institute of Plant Science and Microbiology, Department of Biology, Universität HamburgMolecular Plant Genetics, Institute of Plant Science and Microbiology, Department of Biology, Universität HamburgMolecular Plant Physiology, Institute of Plant Science and Microbiology, Department of Biology, Universität HamburgAbstract Ribosome biogenesis is a key process in all eukaryotic cells that requires hundreds of ribosome biogenesis factors (RBFs), which are essential to build the mature ribosomes consisting of proteins and rRNAs. The processing of the required rRNAs has been studied extensively in yeast and mammals, but in plants much is still unknown. In this study, we focused on a RBF from A. thaliana that we named NUCLEOLAR RNA CHAPERONE-LIKE 1 (NURC1). NURC1 was localized in the nucleolus of plant cell nuclei, and other plant RBF candidates shared the same localization. SEC-SAXS experiments revealed that NURC1 has an elongated and flexible structure. In addition, SEC-MALLS experiments confirmed that NURC1 was present in its monomeric form with a molecular weight of around 28 kDa. RNA binding was assessed by performing microscale thermophoresis with the Arabidopsis internal transcribed spacer 2 (ITS2) of the polycistronic pre-rRNA precursor, which contains the 5.8S, 18S, and 25S rRNA. NURC1 showed binding activity to the ITS2 with a dissociation constant of 228 nM and exhibited RNA chaperone-like activity. Our data suggested that NURC1 may have a function in pre-rRNA processing and thus ribosome biogenesis.https://doi.org/10.1038/s41598-023-36426-4 |
| spellingShingle | Rita Fernandes Anna Ostendorp Steffen Ostendorp Judith Mehrmann Sven Falke Melissa Ann Graewert Magdalena Weingartner Julia Kehr Stefan Hoth Structural and functional analysis of a plant nucleolar RNA chaperone-like protein Scientific Reports |
| title | Structural and functional analysis of a plant nucleolar RNA chaperone-like protein |
| title_full | Structural and functional analysis of a plant nucleolar RNA chaperone-like protein |
| title_fullStr | Structural and functional analysis of a plant nucleolar RNA chaperone-like protein |
| title_full_unstemmed | Structural and functional analysis of a plant nucleolar RNA chaperone-like protein |
| title_short | Structural and functional analysis of a plant nucleolar RNA chaperone-like protein |
| title_sort | structural and functional analysis of a plant nucleolar rna chaperone like protein |
| url | https://doi.org/10.1038/s41598-023-36426-4 |
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