Conditional Modulation of Membrane Protein Expression in Cultured Cells Mediated by Prion Protein Recognition of Short Phosphorothioate Oligodeoxynucleotides

• Published in: The Journal of biological chemistry 2011-03, Vol.286 (9), p.6911-6917
• Main Authors: Karpuj, Marcela Viviana, Gelibter-Niv, Sagit, Tiran, Anat, Rambold, Angelika, Tatzelt, Jörg, Nunziante, Max, Schatzl, Hermann M.
• Format: Article
• Language: English
• Subjects: Animals; Cell Line, Tumor; Cell Membrane - metabolism; CHO Cells; Cricetinae; Cricetulus; Down-Regulation - drug effects; Down-Regulation - physiology; Glycosylation; Membrane Biology; Membrane Proteins; Mice; Neuroblastoma; Oligotherapy; Phosphorothioate Oligonucleotides - pharmacology; Prion Diseases - metabolism; Prion Diseases - therapy; Prions; Protein Degradation; Protein DNA-Interaction; Protein Structure, Tertiary; PrPC Proteins - chemistry; PrPC Proteins - genetics; PrPC Proteins - metabolism; PrPSc Proteins - chemistry; PrPSc Proteins - genetics; PrPSc Proteins - metabolism; RNA, Small Interfering; siRNA; Transfection; Tunicamycin - pharmacology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M110.194662

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.