RNAi screening; p53; 40S; 60S; ribosome; protein kinase; rRNA; image analysis
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Montellese Christian, van den Heuvel Jasmin, Ashiono Caroline, Dörner Kerstin, Melnik André, Jonas Stefanie, Zemp Ivo, Picotti Paola, Gillet Ludovic C, Kutay Ulrike (2020), USP16 counteracts mono-ubiquitination of RPS27a and promotes maturation of the 40S ribosomal subunit, in
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Lukas Bammert, Stefanie Jonas, Rosemarie Ungricht, Ulrike Kutay, Lukas Bammert, Stefanie Jonas, Rosemarie Ungricht, Ulrike Kutay (2016), Human AATF/Che-1 forms a nucleolar protein complex with NGDN and NOL10 required for 40S ribosomal subunit synthesis., in
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Larburu Natacha, Montellese Christian, O'Donohue Marie-Françoise, Kutay Ulrike, Gleizes Pierre-Emmanuel, Plisson-Chastang Célia, Larburu Natacha, Montellese Christian, O'Donohue Marie-Françoise, Kutay Ulrike, Gleizes Pierre-Emmanuel, Plisson-Chastang Célia (2016), Structure of a human pre-40S particle points to a role for RACK1 in the final steps of 18S rRNA processing., in
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The coding information contained in messenger RNAs is translated into proteins by sophisticated ribonucleoprotein particles called ribosomes in all living cells. Ribosomes are composed of two subunits of distinct structure, size, composition and function. The human 80S ribosome, built by the small 40S and the large 60S subunit, is altogether 4.3 MDa in size and composed of 4 ribosomal RNAs (rRNAs) and about 80 ribosomal proteins (RPs). To be functional in protein translation, the ribosomal subunits must be correctly assembled, following a well-ordered pathway during which a long precursor rRNA (pre-rRNA) is processed, modified and folded and the RPs are placed at their designated positions. The synthesis of ribosomal subunits in eukaryotes proceeds along a complex assembly line starting with pre-rRNA synthesis in nucleoli, followed by many intermediate nuclear steps up to their final maturation in the cytoplasm. Proteomic approaches and genetic screens in yeast drove the identification of more than 250 trans-acting factors that assist ribosomal subunit production. These factors associate with pre-ribosomal particles either transiently or as structural components of precursor particles, and are continuously recycled to assist consecutive rounds of ribosome assembly. Although the basic aspects of ribosome biogenesis are conserved between yeast and man, our knowledge on the even more complex process of human ribosome biogenesis is far less complete. This increased complexity not only arises from the larger size of mammalian ribosomes and the more intricate pre-rRNA processing pathway, but also from the highly multifaceted regulatory networks and surveillance pathways linked to ribosome production in vertebrate cells. Since deregulation of ribosome biogenesis manifests in a variety of genetically inherited ribosomopathies and is associated with tumorigenesis, there is great demand to obtain a comprehensive understanding of the process in human cells. As defects in ribosome synthesis activate the p53 stress response pathway leading to cell cycle arrest and apoptosis, factors involved in the making of ribosomes have emerged as attractive targets for cancer therapy.In this grant application, we propose to address some key, unresolved questions related to ribosome biogenesis. As a broad approach, we will further pursue imaging-based RNA interference screening campaigns to comprehensively identify factors involved in human ribosome synthesis. Here, we will give focus on the completion of a genome-wide RNAi screen on 60S subunit synthesis. In parallel, we will exploit data derived from a completed screen on 40S subunit synthesis to specifically address the function of a newly identified protein complex in early steps of pre-rRNA processing, and its role in governing p53 activation. Further, by exploiting the screening data, we will study the recycling of trans-acting factors as well as the role of selected factors in ribosome synthesis. Collectively, this research will not only provide a comprehensive analysis of the machinery involved in human ribosome synthesis, but will also advance the mechanistic understanding of key processes in the making of a fundamental macromolecular complex.