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Etude des mécanismes de régulation de l'épissage des ARN de deux rétrovirus, le virus du Sarcome de Rous (RSV) et le virus responsable de l'immunodéficience humaine (VIH-1)

Abstract : Pre-mRNA splicing is a key step for HIV-1 multiplication. Four donor and eight acceptor sites are used in combination to produce 40 different mRNA. We established the RNA secondary structure around the A7 3' acceptor site and discovered a new regulatory cis acting sequence called Janus element that binds either hnRNP A1 or ASF/SF2 proteins. Moreover, the A7 splicing site is regulated by ASF/SF2, 9G8 and hnRNP A1. We used a proteomic approach based on an affinity chromatography coupled with 1 or 2D gel electrophoresis and mass spectrometry. By this approach, we identified new proteins which can interact with an RNA containing A7 3' acceptor site and the Janus element. Finally, we developed a dual luciferase reporter plasmid that can measure the pre-mRNA splicing efficiency in mammalian cells. In parallel, we established RNA secondary structure of the NRS element of the Rous sarcoma virus and identified the binding sites for SR proteins (9G8, SRp40 and ASF/SF2) and hnRNP A1.
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http://docnum.univ-lorraine.fr/prive/SCD_T_2005_0190_MARCHAND.pdf
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https://hal.univ-lorraine.fr/tel-01746623
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Submitted on : Thursday, March 29, 2018 - 10:41:27 AM
Last modification on : Friday, February 26, 2021 - 3:02:02 PM

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  • HAL Id : tel-01746623, version 1

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Virginie Marchand. Etude des mécanismes de régulation de l'épissage des ARN de deux rétrovirus, le virus du Sarcome de Rous (RSV) et le virus responsable de l'immunodéficience humaine (VIH-1). Biologie moléculaire. Université Henri Poincaré - Nancy 1, 2005. Français. ⟨NNT : 2005NAN10190⟩. ⟨tel-01746623⟩

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