title: Conditional modulation of membrane protein expression in cultured cells mediated by prion protein recognition of short phosphorothioate oligodeoxynucleotides.
abstract: We demonstrate that the levels of native as well as transfected prion protein (PrP) are lowered in various cell lines exposed to phosphorothioate oligodeoxynucleotides (PS-DNA) and can be rapidly reverted to their normal amounts by removal of PS-DNA. This transient modulation was independent of the glycosylation state of PrP, and in addition, all three PrP glycoforms were susceptible to PS-DNA treatment. Deletion of the N-terminal domain (amino acids 23-99), but not of the other domains of PrP, abrogated its PS-DNA-mediated down-regulation. PrP versions localized in the mitochondria, cytoplasm, or nucleus were not modulated by PS-DNA, indicating that PrP surface exposure is required for executing this effect. Proteins that in their native forms were not responsive to PS-DNA, such as thymocyte antigen 1 (Thy1), Doppel protein (Dpl), green fluorescent protein (GFP), and cyan fluorescent protein (CFP), became susceptible to PS-DNA-mediated down-regulation following introduction of the N terminus of PrP into their sequence. These observations demonstrate the essential role of the N-terminal domain for promoting oligonucleotide-mediated reduction of the PrP level and suggest that transient treatment of cultured cells with PS-DNA may provide a general method for targeted modulation of the levels of desired surface proteins in a conditional and reversible manner.
authors.affiliation: Institute of Biochemistry, Food Science and Nutrition Food and Environmental Quality Sciences, Faculty of Agriculture, Hebrew University of Jerusalem, Rehovot 76100, Israel. mvkarpuj@gmail.com
authors.forename: Marcela Viviana
authors.initials: MV
authors.lastname: Karpuj
authors.forename: Sagit
authors.initials: S
authors.lastname: Gelibter-Niv
authors.forename: Anat
authors.initials: A
authors.lastname: Tiran
authors.forename: Angelika
authors.initials: A
authors.lastname: Rambold
authors.forename: Jörg
authors.initials: J
authors.lastname: Tatzelt
authors.forename: Max
authors.initials: M
authors.lastname: Nunziante
authors.forename: Hermann M
authors.initials: HM
authors.lastname: Schatzl
mesh_terms: D000818:Animals; D016466:CHO Cells; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D015536:Down-Regulation / Q000187:drug effects / Q000502:physiology; D006031:Glycosylation; D051379:Mice; D009447:Neuroblastoma; D054735:Phosphorothioate Oligonucleotides / Q000494:pharmacology*; D018621:PrPC Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018620:PrPSc Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017096:Prion Diseases / Q000378:metabolism* / Q000628:therapy; D017434:Protein Structure, Tertiary; D034741:RNA, Small Interfering; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology
chemical_list: D054735:Phosphorothioate Oligonucleotides; D018621:PrPC Proteins; D018620:PrPSc Proteins; D034741:RNA, Small Interfering; D014415:Tunicamycin