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Modifications structurales du Virus de la Mosaïque du Brome et interactions entre particules virales en solution : application à la cristallisation

Abstract : Viruses are biological objects, which mechanisms of proliferation remain badly understood, and which size and organization of the genome make crystallization difficult. We studied the structural modifications of a spherical plant virus, the Brome mosaic Virus (BMV), as well as the interactions between viral particles in solution in order to define crystallization conditions. The capsid swelling that occurs when the virus enters into the host cells is involved in the viral proliferation. We modeled the BMV capsid, in its compact form at pH 5.9 and in its swollen form at pH 7.5, by three-dimensional reconstruction from electron cryomicroscopy images. We thus could observe the rearrangement of RNA between the two forms. We then studied the interactions between viral particles in solution by small angle X-ray scattering. We varied several physicochemical parameters as pH, and polymer and salt concentration, to induce attractive interactions between viruses in solution. Indeed, crystallization of protein occurs in attractive regime. Owing to the results obtained by using polyethylene glycol (PEG), we could determine crystallization conditions of BMV, and show the correlation existing between the nature of the interactions in solution and the crystallization of macromolecules of the size of BMV. We also highlighted that an excess of PEG causes the microcrystalline precipitation of the viruses. The study of kinetics of appearance and growth of the microcrystals enabled us to better characterize the early stages of crystallization of BMV in presence of PEG.
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Submitted on : Friday, June 18, 2004 - 1:19:32 PM
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  • HAL Id : tel-00006118, version 1

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Marina Casselyn. Modifications structurales du Virus de la Mosaïque du Brome et interactions entre particules virales en solution : application à la cristallisation. Biochimie [q-bio.BM]. Université Pierre et Marie Curie - Paris VI, 2002. Français. ⟨tel-00006118⟩

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