Small, Highly Structured RNAs Participate in the Conversion of Human Recombinant PrPSen to PrPRes in Vitro
| Autor: | Valentin Kryukov, Brian Zeiler, Richard Rubenstein, Richard J. Kascsak, Victor Adler, Abraham Grossman |
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| Rok vydání: | 2003 |
| Předmět: |
PrPSc Proteins
Cell law.invention Mice Structural Biology law Cricetinae medicine Animals Humans PrPC Proteins Ribonuclease Nucleic acid structure Molecular Biology Mice Knockout biology Tissue Extracts Brain RNA Ribonuclease Pancreatic Proteinase K Recombinant Proteins In vitro Rats Nucleoprotein Nucleoproteins medicine.anatomical_structure Biochemistry biology.protein Recombinant DNA Nucleic Acid Conformation Protein Binding |
| Zdroj: | Journal of Molecular Biology. 332:47-57 |
| ISSN: | 0022-2836 |
| DOI: | 10.1016/s0022-2836(03)00919-7 |
| Popis: | We have identified a small, highly structured (shs)RNA that binds human recombinant prion protein (hrPrP) with high affinity and specificity under physiological conditions (e.g. 10% bovine calf serum (BCS), neutral pH, nanomolar concentrations of RNA and hrPrP). We also demonstrate the ability of this shsRNA to form highly stable nucleoprotein complexes with hrPrP and cellular PrP (PrP(C)) from various cell extracts and mammalian brain homogenates. The apparent mass of the nucleoprotein complex is dependent on the molar ratio of hrPrP to RNA during complex formation. The hrPrP in these complexes acquires resistance to degradation by Proteinase K (PK). Other shsRNAs, however, under identical conditions, neither form stable complexes with hrPrP nor do they induce resistance to PK digestion. We also demonstrate that the RNAs in these nucleoprotein complexes become resistant to ribonuclease A hydrolysis. These interactions between shsRNAs and hrPrP suggest possible roles of RNAs in the modulation of PrP structure and perhaps disease development. ShsRNAs that bind to hrPrP with high affinity and induce resistance to PK digestion can be used to develop molecular biology assays for the screening of compounds associated with PrP structure transformation or for drugs that inhibit this process. |
| Databáze: | OpenAIRE |
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