title: Nervous and nonnervous cell transduction by recombinant adenoviruses that inducibly express the human PrP.
abstract: The study of the prion protein (PrP) physiological functions or its specific role in transmissible spongiform encephalopathies (TSE) requires new tools, particularly those able to induce PrP overexpression in a large range of cells, in vivo as well as in vitro. Here we describe the construction of two recombinant adenoviruses encoding the human PrP either with a valine at position 129 (AdTRVal) or a methionine (AdTRMet). Both genes were put under the control of the tetracycline-responsive promoter, allowing tight regulation of PrP expression. AdTRVal and AdTRMet induced high expression of the human PrP in CHO-KI cells and in organotypic brain slices in culture. The proteins expressed from these viruses exhibited a glycosylphosphatidyl inositol (GPI) anchor, proper glycosylation and sensitivity to proteinase K digestion. AdTRVal and AdTRMet will allow future studies on the human PrP and on the role of the codon 129 polyphormism in human TSE.
authors.affiliation: CEA, Service de Neurovirologie, BP 6 92265 Fontenay-aux-Roses Cedex, France.
authors.forename: S
authors.initials: S
authors.lastname: Arrabal
authors.forename: M
authors.initials: M
authors.lastname: Touchard
authors.forename: F
authors.initials: F
authors.lastname: Mouthon
authors.forename: B
authors.initials: B
authors.lastname: Klonjkowski
authors.forename: J P
authors.initials: JP
authors.lastname: Deslys
authors.forename: D
authors.initials: D
authors.lastname: Dormont
authors.forename: M
authors.initials: M
authors.lastname: Eloit
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D019943:Amino Acid Substitution; D000818:Animals; D016466:CHO Cells / Q000166:cytology / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002531:Cerebellum / Q000378:metabolism; D006224:Cricetinae; D004318:Doxycycline / Q000494:pharmacology; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006624:Hippocampus / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D018345:Mice, Knockout; D016297:Mutagenesis, Site-Directed; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011328:Prions / Q000096:biosynthesis* / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D013752:Tetracycline / Q000494:pharmacology; D014161:Transduction, Genetic / Q000379:methods*
chemical_list: D011328:Prions; D012333:RNA, Messenger; D013752:Tetracycline; D004318:Doxycycline