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Communication Dans Un Congrès Année : 2007

Glycan chain modifications and prion propagation in cultured cells

Résumé

Rov cells do not express endogenous PrPc and are infectible by sheep scrapie prions upon expression of ovine PrPc, therefore providing a useful reverse genetic system to study effects of sequence modifications on the prion infection process. To further investigate how the glycan chains influence the conversion into PrPSc and the prion propagation, we introduced various mutations that modify the glycosylation state of the molecule in different ways, and analysed pools of selected Rov cell clones for the accumulation of PrPSc upon serial passage after exposure to 127S strain infectious prion. In a first series of constructs, either one or both asparagines at position 184 and 200 (sheep sequence numbering) were replaced by a set of amino acids to create monoor non-glycosylated mutants. The results available to date showed that: i) monoglycosylated mutants at either site can confer permissiveness to infection, however, the accumulation of PrPSc can vary dramatically according to the amino acid substitution, despite a cell surface expression of the mutated PrPc at similar levels; ii) the distal site (200) appeared to be more tolerant to mutation than the proximal site for permissiveness to infection, reminiscent of that reported for mouse PrP in transgenic mice1; iii) double mutants exhibited an impaired subcellular trafficking with overaccumulation in the Golgi compartment, and failed to be infected as yet. Altogether the above results raised the possibility that either or both amino acid change at the glycan attachment site and altered cell surface expression, rather than the ablation of the glycan chain itself, may impair the infection process. This incited us to undertake the production of a second generation of constructs, including the suppression of the GPI anchor2, that may led to the production of PrPc molecules with altered glycosylation without a substitution within the glycosylation sequon, a work that is currently in progress.
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Dates et versions

hal-02751281 , version 1 (03-06-2020)

Identifiants

  • HAL Id : hal-02751281 , version 1
  • PRODINRA : 147844

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Michel Dron, Christelle C. Langevin, Jerome Chapuis, Didier Vilette, Mohammed M. Moudjou, et al.. Glycan chain modifications and prion propagation in cultured cells. Prion 2007, Sep 2007, Edinburgh, United Kingdom. ⟨hal-02751281⟩
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