In situ photodegradation of incorporated polyanion does not alter prion infectivity.
Single-stranded polyanions ≥40 bases in length facilitate the formation of hamster scrapie prions in vitro, and polyanions co-localize with PrP(Sc) aggregates in vivo. To test the hypothesis that intact polyanionic molecules might serve as a structural backbone essential for maintaining the infectio...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2011-02-01
|
| Series: | PLoS Pathogens |
| Online Access: | http://europepmc.org/articles/PMC3033378?pdf=render |
| _version_ | 1818506827322621952 |
|---|---|
| author | Justin R Piro Brent T Harris Surachai Supattapone |
| author_facet | Justin R Piro Brent T Harris Surachai Supattapone |
| author_sort | Justin R Piro |
| collection | DOAJ |
| description | Single-stranded polyanions ≥40 bases in length facilitate the formation of hamster scrapie prions in vitro, and polyanions co-localize with PrP(Sc) aggregates in vivo. To test the hypothesis that intact polyanionic molecules might serve as a structural backbone essential for maintaining the infectious conformation(s) of PrP(Sc), we produced synthetic prions using a photocleavable, 100-base oligonucleotide (PC-oligo). In serial Protein Misfolding Cyclic Amplification (sPMCA) reactions using purified PrP(C) substrate, PC-oligo was incorporated into physical complexes with PrP(Sc) molecules that were resistant to benzonase digestion. Exposure of these nuclease-resistant prion complexes to long wave ultraviolet light (315 nm) induced degradation of PC-oligo into 5 base fragments. Light-induced photolysis of incorporated PC-oligo did not alter the infectivity of in vitro-generated prions, as determined by bioassay in hamsters and brain homogenate sPMCA assays. Neuropathological analysis also revealed no significant differences in the neurotropism of prions containing intact versus degraded PC-oligo. These results show that polyanions >5 bases in length are not required for maintaining the infectious properties of in vitro-generated scrapie prions, and indicate that such properties are maintained either by short polyanion remnants, other co-purified cofactors, or by PrP(Sc) molecules alone. |
| first_indexed | 2024-12-10T22:09:53Z |
| format | Article |
| id | doaj.art-a62dfa2a40da43d4ab6cb4a9073652ef |
| institution | Directory Open Access Journal |
| issn | 1553-7366 1553-7374 |
| language | English |
| last_indexed | 2024-12-10T22:09:53Z |
| publishDate | 2011-02-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj.art-a62dfa2a40da43d4ab6cb4a9073652ef2022-12-22T01:31:38ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742011-02-0172e100200110.1371/journal.ppat.1002001In situ photodegradation of incorporated polyanion does not alter prion infectivity.Justin R PiroBrent T HarrisSurachai SupattaponeSingle-stranded polyanions ≥40 bases in length facilitate the formation of hamster scrapie prions in vitro, and polyanions co-localize with PrP(Sc) aggregates in vivo. To test the hypothesis that intact polyanionic molecules might serve as a structural backbone essential for maintaining the infectious conformation(s) of PrP(Sc), we produced synthetic prions using a photocleavable, 100-base oligonucleotide (PC-oligo). In serial Protein Misfolding Cyclic Amplification (sPMCA) reactions using purified PrP(C) substrate, PC-oligo was incorporated into physical complexes with PrP(Sc) molecules that were resistant to benzonase digestion. Exposure of these nuclease-resistant prion complexes to long wave ultraviolet light (315 nm) induced degradation of PC-oligo into 5 base fragments. Light-induced photolysis of incorporated PC-oligo did not alter the infectivity of in vitro-generated prions, as determined by bioassay in hamsters and brain homogenate sPMCA assays. Neuropathological analysis also revealed no significant differences in the neurotropism of prions containing intact versus degraded PC-oligo. These results show that polyanions >5 bases in length are not required for maintaining the infectious properties of in vitro-generated scrapie prions, and indicate that such properties are maintained either by short polyanion remnants, other co-purified cofactors, or by PrP(Sc) molecules alone.http://europepmc.org/articles/PMC3033378?pdf=render |
| spellingShingle | Justin R Piro Brent T Harris Surachai Supattapone In situ photodegradation of incorporated polyanion does not alter prion infectivity. PLoS Pathogens |
| title | In situ photodegradation of incorporated polyanion does not alter prion infectivity. |
| title_full | In situ photodegradation of incorporated polyanion does not alter prion infectivity. |
| title_fullStr | In situ photodegradation of incorporated polyanion does not alter prion infectivity. |
| title_full_unstemmed | In situ photodegradation of incorporated polyanion does not alter prion infectivity. |
| title_short | In situ photodegradation of incorporated polyanion does not alter prion infectivity. |
| title_sort | in situ photodegradation of incorporated polyanion does not alter prion infectivity |
| url | http://europepmc.org/articles/PMC3033378?pdf=render |
| work_keys_str_mv | AT justinrpiro insituphotodegradationofincorporatedpolyaniondoesnotalterprioninfectivity AT brenttharris insituphotodegradationofincorporatedpolyaniondoesnotalterprioninfectivity AT surachaisupattapone insituphotodegradationofincorporatedpolyaniondoesnotalterprioninfectivity |