Utilisation du facteur bactérien Rho pour l'étude de la biogénèse et du contrôle qualité des transcrits eucaryotes

Abstract : In eukaryotes, the transcription of genetic information into messenger RNA is a complex process which requires multiple modifications of precursor RNA molecule, termed mRNA processing. Current view is that the nascent RNA is sequentially coated with a large set of proteins to generate messenger ribonucleoprotein particles (mRNPs). Various mRNA processing and assembly events, known as mRNP biogenesis, are coupled physically and functionally to transcription. They confer integrity and promote export of transcript to the cytoplasm for translation. The production of mature export-competent transcripts is interconnected with a quality control step in order to avoid the export of improperly processed transcript. To identify new factors of mRNP biogenesis and understand the signalisation mode of aberrant transcripts, we have implemented an innovative screen. This screen is based on the perturbation of gene expression in Saccharomyces cerevisiae by bacterial Rho factor which is able to disrupt proteins interacting with RNA molecule in vitro. Rho expression in yeast interferes with co-transcriptional nucleoprotein assembly and generates aberrant transcripts which are degraded by the nuclear surveillance system. This study reveals dynamic interactions of protein factors with transcriptional complex and their implications in recognition of aberrant transcripts by the degradation machinery. Our methodology opens interesting prospects to detect new mRNP biogenesis factors and value their role in regulation of genes expression.
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Romy Honorine. Utilisation du facteur bactérien Rho pour l'étude de la biogénèse et du contrôle qualité des transcrits eucaryotes. Sciences agricoles. Université d'Orléans, 2010. Français. ⟨NNT : 2010ORLE2004⟩. ⟨tel-00558795⟩



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