Eukaryotic Ribosome Biogenesis: Focus on the function of the assembly factor Mrd1p
2012 (English)Doctoral thesis, comprehensive summary (Other academic)
The ribosome, the protein factory of the cell, is essential for all life forms. The ribosome is a large RNA-protein machine. It is built in a complex, multi-step process that involves a large number of accessory trans-acting factors and the synthesis consumes a considerable part of the cellular energy. The ribosomal RNA is transcribed as a large precursor rRNA (pre-rRNA) molecule that undergoes extensive processing during maturation, including chemical modifications, pre-rRNA cleavage events, pre-rRNA folding and assembly with ribosomal proteins. More than 200 non-ribosomal proteins and small nucleolar RNAs ensure a successful maturation of the two ribosomal subunits during a pathway that starts with coupled synthesis and processing of the pre-rRNA within the nucleolus. Processing continues through the nucleus and ends with the final maturation in the cytoplasm.
We have studied one of the eukaryotic ribosomal biogenesis proteins, Mrd1 to learn about its essential function in the pre-ribosome maturation process in the yeast, Saccharomyces cerevisiae. Mrd1 contains multiple RNA-binding domains and the protein and its modular design is conserved throughout eukarya. Evolution of Mrd1 is most likely coupled to a common eukaryotic way of producing ribosomes. Together with a large set of other factors, Mrd1 associates early with the nascent pre-rRNA and forms a 90S pre-ribosome that can be seen in Chromatin Miller spreads of active rRNA genes as large terminal knob structures on the growing pre-rRNA. In the absence of Mrd1, essential steps in pre-ribosome maturation cannot occur and small ribosomal subunits are not produced. We have demonstrated that Mrd1 interacts with the pre-rRNA in vivo at two specific sites within the 18S rRNA sequence, both located close in space to where the essential and universally conserved central pseudoknot of the small ribosomal subunit is formed. Furthermore, we have shown that Mrd1 influences the release of the U3 snoRNA from the pre-ribosome. U3 snoRNA is essential for synthesis of the small ribosomal subunit and is involved in pseudoknot formation. Our results show that Mrd1 is present within the pre-ribosome at a crucial location and that it is required for essential maturation steps. Based on our results, we hypothesize that Mrd1 modulates the pre-rRNA folding and assembly to assist pre-ribosome structures necessary for pseudoknot formation and early cleavages. This essential function is conserved in all eukaryotes.
Place, publisher, year, edition, pages
Stockholm: Department of Molecular Biology and Functional Genomics, Stockholm University , 2012. , 83 p.
Ribosome biogenesis, nucleolus, pre-rRNA processing, RNA-binding proteins, Mrd1
Research subject Molecular Biology
IdentifiersURN: urn:nbn:se:su:diva-75829ISBN: 978-91-7447-509-8OAI: oai:DiVA.org:su-75829DiVA: diva2:524227
2012-06-07, De Geersalen, Geovetenskapens hus, Svante Arrhenius väg 14, Stockholm, 10:00 (English)
Gerbi, Susan, Professor
Wieslander, Lars, Professor
At the time of doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Manuscript. Paper 4: Manuscript. 2012-05-102012-04-302012-05-03Bibliographically approved
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